PARP1 inhibitors and the use in the treatment of CNS diseases

Abstract

The invention provides PARP1 inhibitors and their use in the treatment of CNS diseases. The PARP1 inhibitor of the invention has the structure shown in Formula (I). Compared to PARP2 activity, the PARP1 inhibitor can selectively inhibit PARP1 activity. The invention also provides the use of the PARP1 inhibitor in the treatment of CNS diseases.

Description

064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS PARP1 INHIBITORS AND THE USE IN THE TREATMENT OF CNS DISEASES Field of the Invention

[0001] The invention is in the field of medicinal chemistry, and relates to the field of disease treatment. In particular, the invention relates to the use of PARP1 inhibitors in the treatment of CNS diseases, such as brain tumors. Background of the Invention

[0002] Poly(ADP-ribose) polymerase (PARP) is a family of proteins, which transfer negatively charged ADP-ribose groups from donor NAD+onto target proteins. That is one of many post-transcriptional modifications. Therefore, PARP is also termed an ADP-ribose transferase.

[0003] The most characteristic member of the PARP family is PARP1, which was found to have the highest intracellular levels. PARP1 consists of 1014 amino acids (NCBI Accession P09874) with a total molecular weight of approximately 116 kDa. Structurally, this enzyme is composed of two main domains including an N-terminal DNA-binding domain and a catalytic domain. PARP1 is known to play an important role in many cellular functions, including gene expression, transcription, cell division, cell differentiation, cell apoptosis, DNA damage response and repair. PARP1 is activated when DNA damage occurs and is involved in base excision repair (BER) which is a major mechanism of DNA single-strand damage repair. PARP1 binds to the site of Single Strand Break (SSB), and subsequently repair DNA via BER. In response to DNA damage, cells also have evolved two main repair pathways: Homologous Recombination (HR) and Non-Homologous End Joining (NHEJ), in addition to BER repair mechanisms. HR deficient tumors have been found to be sensitive to PARP inhibitors, indicating homologous recombination defects and PARP1 inhibition formed a pair of synthetic lethality, which has been validated by clinical studies. Several PARP inhibitors are currently approved for the treatment of ovarian, breast, pancreatic and prostate cancers with DNA damage repair deficient such as BRCA1 / 2 mutation. In addition, PARP1 inhibitors may also be used to treat diseases due to excessive cell death, including central nervous system (CNS) diseases such as stroke and neurodegenerative diseases (Akinori Iwashita et al., 2004, J. Pharmacol. Exp. Thera.310: 425; Marianna Mekhaeil et al., 2023, Neurotherapeutics 20: 1347- 1368).064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0004] PARP2 is a protein of 583 amino acids with molecular mass of approximately 62 kDa and composed of DNA binding domain and catalytic domain (Ame et al., 1999 J Biol Chem 274: 17860-17868). The catalytic domain of PARP2 is very similar to that of PARP1. PARP2 is also found to have similar functions to PARP1 and is involved in the repair of DNA damage through BER mechanism (Schreiber et al., 2002 J Biol Chem 277: 23028-23036). Marketed PARP inhibitors, such as Olaparib, Niraparib, Talazoparib and Rucaparib, not only have inhibitory activities against PARP1, but also have similar inhibitory activities against PARP2. Based on the results of clinical trials, the therapeutic effects of these marketed PARP inhibitors are comparable whereas their toxicity profiles are quite different. For example, Talazoparib has toxicity similar to chemotherapy drugs such as hair loss. A recent study showed that Talazoparib, in addition to inhibiting PARP1 and PARP2, also has a fairly high affinity for TNKS1 (Tankyrase 1) and TNKS2 (Tankyrase 2), two other members of the PARP family (Ryan et al., 2021 J Biol Chem 296: 100251 / 1-100251 / 13). TNKS1 and TNKS2 share 83% sequence identity overall, and their catalytic domain sequences are 89% identical. They play roles in DNA repair, telomere maintenance, and Wnt / β-catenin signaling. Targeting PARPs other than PARP1 may be the reason why PARP inhibitors cause off-targeted toxicity, such as hair loss and diarrhea. In addition, inhibition of PARP2 activity has been found to lead to hematotoxicity (Farrés et al., 2013 Blood 122: 44-54; Farrés et al., 2015 Cell Death and Differentiation 22: 1144-1157). The toxicity of these PARP inhibitors limits their clinical application as well as in combination with other targeted drugs.

[0005] Central nervous system diseases, also known as central nervous system disorders, are a group of neurological disorders that affect the structure or function of the brain or spinal cord, which collectively form the central nervous system (CNS). Brain tumors are the most frequent and have the highest mortality (Daroff et al., 2014 Encyclopedia of the Neurological Sciences, eBook ISBN: 9780123851581).

[0006] Brain tumor is considered one of the deadliest cancers in the world. The unique tumor microenvironment and inherent characteristics of brain tumor cells make them resistant to most conventional and advanced therapies. Given the poor survival rates of current brain tumor treatment strategies, new treatment strategies are urgently needed.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Summary of the Invention

[0007] In an aspect, the present disclosure provides a compound having the following structure of Formula (I): (I) or a hydrate, or anacceptable salt thereof, wherein: R1is selected from the group of halo, alkyl and haloalkyl, R2is selected from the group of halo and alkyl, R3 is selected from the group of halo, alkyl and cyano (that is, –CN), and R4is selected from the group of alkyl, haloalkyl and cycloalkyl.

[0008] In some embodiments of the compound of Formula (I) disclosed herein and of the methods and uses disclosed herein, (a) R1 is halo, C1-3 alkyl or halo C1-3 alkyl, or (b) R2 is halo or C1-3alkyl, or (c) R3is halo, C1-3alkyl or cyano, or (d) R4is C1-3alkyl, halo C1-3alkyl or C3-4 cycloalkyl, or any combination of (a) through (d), for example all of (a) through (d).

[0009] In some embodiments of the compound of Formula (I) disclosed herein and of the methods and uses disclosed herein, (e) R1 is F, methyl or trifluoromethyl, or (f) R2 is F, Cl or methyl, or (g) R3is F, methyl or cyano, or (h) R4is C1-3alkyl, C1-3haloalkyl or C3-4cycloalkyl, or any combination of (e) through (h), for example all of (e) through (h).

[0010] In some embodiments of the compound of Formula (I) disclosed herein and of the methods and uses disclosed herein, (i) R1is F, methyl and trifluoromethyl, or (j) R2is F, Cl or methyl, or (k) R3 is F, methyl or cyano, or (l) R4 is methyl, ethyl, difluoroethyl or cyclopropyl, or any combination of (i) through (l), for example all of (i) through (l).

[0011] In some embodiments of the compound of Formula (I): R1is defined as in any one of (a), (e) and (i), above, R2 is defined as in any one of (b), (f) and (j), above, R3 is defined as in any one of (c), (g) and (k), above, and R4is defined as in any one of (d), (h) and (l), above.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0012] In some embodiments of the compound of Formula (I) disclosed herein and of the methods and uses disclosed herein, R1 is F, CH3 or CF3; R2 is F, Cl or CH3; R3 is F, CH3 or cyano; and R4is C1-3alkyl, C1-3haloalkyl or C3-4cycloalkyl. In some embodiments, R1is F, R2is F, R3 is halo, or alkyl, and R4 is alkyl, haloalkyl or cycloalkyl. In some embodiments, R1 is CH3, R2is F, R3is halo, alkyl or cyano, and R4is alkyl, haloalkyl or cycloalkyl. In some embodiments, all of R1, R2, R3 and R4 of the compound are defined according to any one of rows 1-11 of the following table: R1R2R3R41 CH3F F CH32 CH3F CN CH33 CH3F F cyclopropyl 4 CF3 F F CH3 5 F F CH3 CH3 6 F F F CH3 7 F F F CH2CH3 8 F F F CH2CHF2 9 F F F cyclopropyl 10 F Cl F CH3 11 F CH3F CH3

[0013] In some embodiments of the compounds and of the methods disclosed herein, R1 is CH3, R2 is F, R3 is F, and R4 is CH3. In some embodiments of the compounds and methods disclosed herein, R1 is CH3, R2 is F, R3 is CN, and R4 is CH3. In some embodiments of the compounds and methods disclosed herein, R1 is CH3, R2 is F, R3 is F, and R4 is cyclopropyl. In some embodiments of the compounds and methods disclosed herein, R1 is CH3, R2 is F, R3 is F, and R4 is CH3. In some embodiments of the compounds and methods disclosed herein, R1 is F, R2 is F, R3 is CH3, and R4 is CH3. In some embodiments of the compounds and methods disclosed herein, R1 is F, R2 is F, R3 is F, and R4 is CH3. In some embodiments of the compounds and methods disclosed herein, R1is F, R2is F, R3is F, and R4is CH2CH3. In some embodiments of the compounds and methods disclosed herein, R1 is F, R2 is F, R3 is F, and R4 is CH2CF2H. In some embodiments of the compounds and methods disclosed herein, R1is F, R2 is F, R3 is F, and R4 is cyclopropyl. In some embodiments of the compounds and methods064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS disclosed herein, R1is F, R2is Cl, R3is F, and R4is CH3. In some embodiments of the compounds and methods disclosed herein, R1 is F, R2 is CH3, R3 is F, and R4 is CH3.

[0014] In an aspect, the present disclosure provides a process for preparing any of the compounds disclosed herein. In an exemplary embodiment, the present disclosure provides a process for preparing 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1- yl)methyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one comprising: (a) reacting 4-fluoro-1H-pyrazole-5-carboxylic acid and an acylation reagent (such as SOCl2) to produce 4-fluoro-1H-pyrazole-5-carbonyl chloride; (b) reacting 4-fluoro-1H-pyrazole-5-carbonyl chloride and 3,5-dibromo-2,6- difluoroaniline under alkaline condition (such as under the catalysis of NaH, NaHMDS or LiHMDS (lithium bis(trimethylsilyl)amide)) to produce N-(3,5-dibromo-2,6-difluorophenyl)- 4-fluoro-1H-pyrazole-5-carboxamide; (c) ring closing of N-(3,5-dibromo-2,6-difluorophenyl)-4-fluoro-1H-pyrazole-5- carboxamide under alkaline condition (such as under the catalysis of K2CO3) to produce 7,9- dibromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one; (d) selective dehalogenation of 7,9-dibromo-3,6-difluoropyrazolo[1,5-a]quinoxalin- 4(5H)-one to produce 7-bromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one; (e) reacting 7-bromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one and (Bu)3SnCH2OH under the catalysis of Pd catalyst (such as Xphos Pd G2) to produce 3,6- difluoro-7-(hydroxymethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one; (f) reacting 3,6-difluoro-7-(hydroxymethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one and HBr to produce 7-(bromomethyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one; (g) reacting 7-(bromomethyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one and 6-fluoro-N-methyl-5-(piperazin-1-yl)picolinamide under alkaline condition (such as under the catalysis of DIEA and KI) to produce 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3- yl)piperazin-1-yl)methyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one.

[0015] In an aspect, the present disclosure provides a method for inducing regression of a central nervous system tumor in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, tumor regression is induced. In some embodiments,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS inducing regression of the tumor is evaluated from 7 to 84 days after administering the compound e.g. any intervening integer number of days after administering the compound, e.g. 8 days after, 9 days after, etc. or within any range between any two such values. In some embodiments, tumor regression is between 1% and 100%, between 5% and 100%, between 10% and 100%, between 25% and 100%, between 50% and 100%, between 75% and 100%, or is any percentage value between 1% and 100%, e.g. 2%, 3%, 4%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% etc. or within any range between any two such values. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments, the CNS tumor, for example, a brain tumor, has a volume at time t that is smaller than the tumor’s volume on the first day of administering the compound and time t is any of day 7, day 84, or any day between day 7 and day 84. In some embodiments of the method for inducing regression of a central nervous system tumor in a subject, the central nervous system tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumour. In some embodiments of the method for inducing regression of a central nervous system tumor in a subject, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments of the method for inducing regression of a central nervous system tumor in a subject, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0016] In an aspect, the present disclosure provides a method for inducing increased regression of a central nervous system tumor in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, increased tumor regression is induced. In some embodiments, inducing increased regression of the tumor is evaluated from 7 to 84 days after administering the compound e.g. any intervening integer number of days after administering the compound, e.g. 8 days after, 9 days after, etc. or within any range between any two such values. In some embodiments, tumor regression is between 1% and 100%, between 5% and 100%, between 10% and 100%, between 25% and 100%, between 50% and 100%, between 75% and 100%, or is any percentage value between 1% and 100%, e.g. 2%, 3%, 4%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% etc. or within any range between any two such values. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments of the method for inducing increased regression of a central nervous system tumor in a subject, the central nervous system tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumor. In some embodiments of the064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS method for inducing increased regression of a central nervous system tumor in a subject, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments of the method for inducing increased regression of a central nervous system tumor in a subject, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope- substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0017] In an aspect, the present disclosure provides a method for inhibiting growth of a central nervous system tumor in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, tumor growth is inhibited. In some embodiments, inhibiting growth of a central nervous system tumor is evaluated from 7 to 84 days after administering the compound, e.g. any intervening integer number of days after administering the compound, e.g. 8 days after, 9 days after, etc. or within any range between any two such values. In some embodiments, tumor growth is inhibited by 10% to 150% e.g. by 30% to 150% or 50% to 120% e.g. any integer percentage value between 10% and 150%, e.g. by 30%, 40%, 50%, 60%, 70%, 100%, 125%, 150% etc. or within any range between any two such values. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments of the method for inhibiting growth of a central nervous system tumor in a subject, the central nervous system tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumor. In some embodiments of the method for inhibiting growth of a central nervous system tumor in a subject, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments of the method for inhibiting growth of a central nervous system tumor in a subject, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0018] In an aspect, the present disclosure provides a method for inhibiting PARP1 activity e.g. selectively inhibiting PARP1 activity relative to PARP2 activity in a subject comprising administering to the subject an effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, inhibition of PARP1 activity is greater than064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS inhibition of PARP2 activity in the subject. In some embodiments, the ratio of PARP1 activity inhibition to PARP2 activity inhibition in the subject is at least 50, at least 750, at least 1500, at least 2500, at least 5,000, at least 7,500, at least 10,000, at least 15,000, at least 20,000, at least 25,000, or within a range between any two of these numbers. In some embodiments, the ratio of PARP1 activity inhibition to PARP2 activity inhibition in the subject is between 50 and 2,000, between 750 and 15,000, between 2500 and 15,000, or between 12,500 and 25,000 e.g. between 15,000 and 25,000. In some embodiments, the subject is a human or is a non- human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, an effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope- substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0019] In an aspect, the present disclosure provides a method for treating a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject has a disease or condition that is characterized by or due to excessive cell death. In some embodiments, the disease or condition is stroke or a neurodegenerative disease.

[0020] In a further aspect, the present disclosure provides a method for treating cancer in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is an ovarian, breast, pancreatic or prostate cancer. In some embodiments, the cancer is deficient in DNA damage repair e.g. the cancer is deficient in homologous recombination. In some embodiments, the subject has a central nervous system (CNS) tumor. In some embodiments,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS the subject has a brain tumor. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments, the subject has a central nervous system (CNS) tumor. In some embodiments, the CNS tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumour. In some embodiments, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I) , or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0021] In an aspect, the present disclosure provides the use of a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for the preparation of a medicament. In an aspect, the present disclosure provides the use of a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for the preparation of a medicament for use in treating any condition e.g. a cancer, such as any CNS tumor, for example, a brain tumor, as disclosed or described herein.

[0022] In an aspect, the present disclosure provides the use of a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for to treat a condition disclosed herein or for a purpose disclosed herein.

[0023] In an aspect, the present disclosure provides a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for use in a method of treating a condition disclosed herein or for use in a purpose disclosed herein.

[0024] In an aspect, the present disclosure provides a method of treating a central nervous system (CNS) disease or condition e.g. a CNS tumor, for example a brain tumor, comprising administering a compound of Formula I as disclosed herein, wherein the R groups are defined as set forth herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) can penetrate the blood brain barrier (BBB). In some embodiments, the compound of Formula (I) penetrates the blood brain barrier (BBB). In some embodiments, the CNS disease or condition is a brain tumor. In some embodiments, the central nervous system (CNS) disease or condition is a CNS tumor. In some embodiments, the central nervous system (CNS) disease or condition is a CNS tumor selected from meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumour. In some embodiments, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments, a therapeutically effective amount of the compound or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered. In some embodiments, the therapeutically effective amount of the compound or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients.

[0025] In an aspect, the present disclosure provides a pharmaceutically acceptable composition comprising a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable diluent, carrier or excipient.

[0026] In some embodiments of the methods disclosed herein, the method comprises administering a therapeutically effective amount of a compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof as described herein. In some embodiments of the methods disclosed herein, the method comprises administering a therapeutically effective amount of a compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, as described herein. In various embodiments, the pharmaceutically acceptable salt is an inorganic or organic acid salt, e.g. a hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate or oxalate salt. In various embodiments, the pharmaceutically acceptable salt is an inorganic or organic base salts formed with bases, e.g. a sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) or N-methyl-glucamine salt.

[0027] In any embodiments of any of the methods and uses disclosed herein wherein a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs or is used in combination with one or more additional anticancer therapies or drugs, the one or more additional anticancer therapies or drugs is selected from, for example,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS monoclonal antibodies, surgery, radiation therapy (such as ionizing radiation (IR), gamma- radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes), endocrine therapy, biologic response modifiers (such as interferons, interleukins, and tumor necrosis factor (TNF)), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, spindle poisons (such as vinblastine, vincristine, vinorelbine, and paclitaxel), podophyllotoxins (such as etoposide, irinotecan, and vopotecan), nitrosoureas (such as varmustine and lomustine), inorganic ions (such as cisplatin and carboplatin), enzymes (such as asparaginase), hormones (such as tamoxifen, leuprolide, flutamide, and megestrol), Gleevec™, adriamycin, dexamethasone, and cyclophosphamide.

[0028] In any of the methods of treatment disclosed herein, in some embodiments, the subject is a subject in need of such treatment. In various embodiments, the subject is a human. In some embodiments, the subject is a non-human such as a veterinary animal (e.g. pig, cow, sheep, horse, dog, and cat) or a zoo or laboratory animal (e.g. any non-human primate or mammal).

[0029] In any of the uses of a compound disclosed herein in the treatment of a condition in a subject, in some embodiments, the subject is a subject in need of such treatment. In various embodiments, the subject is a human. In some embodiments, the subject is a non-human such as a veterinary animal (e.g. pig, cow, sheep, horse, dog, and cat) or a zoo or laboratory animal (e.g. any non-human primate or mammal).

[0030] In any of the uses of a compound disclosed herein for the preparation of a medicament for use in the treatment of a subject or for use in the treatment of a condition in a subject, in some embodiments, the subject is a subject in need of such treatment. In various embodiments, the subject is a human. In some embodiments, the subject is a non-human such as a veterinary animal (e.g. pig, cow, sheep, horse, dog, and cat) or a zoo or laboratory animal (e.g. any non- human primate or mammal). Detailed Description of the Invention

[0031] The term “alkyl” as used herein refers to alkyl group of a straight or branched chain radical of up to ten carbons. Useful alkyl groups include straight-chain or branched C1-10alkyl groups, for example C1-6 alkyl groups. In some embodiments, alkyl is C1-3 alkyl. Typical alkyl groups include methyl, deuterated methyl ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS pentyl (such as 3-pentyl), hexyl and octyl groups, which may be optionally substituted. In some embodiments, alkyl groups e.g. C1-3 alkyl and C1-6 alkyl groups are unsubstituted. In some embodiments, alkyl groups e.g. C1-3alkyl and C1-6alkyl groups are substituted. In some embodiments, substituents of alkyl groups include halogen groups (F, Cl, Br, and I), —OH, —CHO, —NH2, or a combination thereof.

[0032] The term “cycloalkyl” as used herein refers to cyclic alkyl groups. Useful cycloalkyl groups are C3-8 cycloalkyl. In some embodiments, cycloalkyl groups are C3-6 cycloalkyl. Typical cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. Cycloalkyl group may be substituted by one or more substituents as described herein. In some embodiments, cycloalkyl groups e.g. C3-4cycloalkyl and C3-6cycloalkyl groups are unsubstituted.

[0033] Useful halo or halogen groups include fluoro (F), chloro (Cl), bromo (Br) and iodo (I).

[0034] The term “haloalkyl” as used herein refers to alkyl groups in which one or more H atoms is replaced with one or more halo atoms. In some embodiments, haloalkyl groups are halo C1-3 alkyl. Exemplary haloalkyl include trifluoromethyl, difluoroethyl, fluoromethyl, and dichloroethyl.

[0035] The term “induce” and grammatical variations thereof (e.g. “induction”) as used herein means “to cause” or “to bring about.”

[0036] The term “inhibition” includes a reduction in a certain parameter, e.g., tumor growth. For example, inhibition of tumor growth after exposure to a compound disclosed herein indicates that tumor growth is reduced by, for example, 5%, 10%, 20%, 30%, 40% or more relative to a comparable tumor not exposed to the compound. Inhibition can be less than, equal to, or greater than 100%. If exposure of a subject to a compound results in a reduction in a parameter, e.g. tumor growth, then that compound is an “inhibitor” of the parameter, e.g. tumor growth.

[0037] The term “regression” and grammatical variations thereof (e.g. the verb form “to regress”) as used herein refers to a decrease in the size or extent of a tumor. Such regression may be associated with the disappearance of, or decrease in number or size of, tumor cells (e.g. neoplastic cells).064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0038] The term “inducing tumor regression” and grammatical variations thereof therefore refers to causing or bringing about a decrease in the size or extent of a tumor (including to cause the disappearance (e.g. to the point of undetectability) of the tumor), e.g. by causing the disappearance of or a decrease in the number or size of tumor cells (e.g. neoplastic cells).

[0039] The term “inducing increased regression” refers to causing regression that is already in progress to proceed at a greater rate or to a greater extent, or to causing additional regression where prior regression has ceased. Inducing increased regression of a tumor thus refers to circumstances where, for example, after treatment with, for example, a compound of Formula (I) of the present disclosure, a tumor that is regressing is caused to regress at a greater rate or to a greater extent, or if the tumor has previously ceased to regress, additional regression is caused by, for example, treatment with, for example, a compound of Formula (I) of the present disclosure. Inducing increased regression can occur when e.g. administration of two different (e.g. independently acting) treatments both cause regression and, combined, they provide an additive or synergistic effect.

[0040] The term “pharmaceutically acceptable,” as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Accordingly, pharmaceutically acceptable relates to substances that are not biologically or otherwise undesirable, i.e., the material can be administered to an individual along with the relevant active compound without causing clinically unacceptable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained.

[0041] As used herein, the term “therapeutically effective amount” or “effective amount” of a therapeutic agent, or of a composition comprising a therapeutic agent, means an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of one or more symptom(s) of the disease, disorder, and / or condition (e.g. slow, or prevent, an increase in severity of any such symptoms, or reduce the severity of any such symptoms). “Therapeutically effective amount” and “effective amount” also encompass an amount that is sufficient, when administered to a subject, to impact an activity or result in a change in an activity, such as an enzymatic activity such as PARP1 activity or PARP2 activity. It will be appreciated by those064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS of ordinary skill in the art that a therapeutically effective amount or effective amount is typically administered via a dosing regimen comprising at least one unit dose. A non-limiting example of a “therapeutically effective amount” or an "effective amount" of the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the composition comprising the compound of Formula (I) of the present disclosure is an amount that can be used to inhibit, block or reverse the activation, migration or proliferation of cells, or to effectively treat cancer or improve symptoms of cancer. A further non-limiting example of a “therapeutically effective amount” or an "effective amount" of the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the composition comprising the compound of Formula (I) of the present disclosure is an amount that can be used to inhibit the activity of PARP1.

[0042] As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of and / or reduce incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and / or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease.

[0043] Tumor cells (or Cancer cells) described herein are generally characterized by abnormal proliferation relative to normal cells and the formation of clusters or tumors in individuals suffering from cancer. “Cancer” includes a disease characterized by the rapid and uncontrolled growth of aberrant cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers include cancers of the central nervous system, for example, brain cancers and spinal cord cancer. The terms “tumor” and “cancer” are used interchangeably herein. For example, both terms encompass solid and liquid, e.g., diffuse or circulating tumors. In some embodiments, the term “cancer” or “tumor” includes malignant cancers and tumors, as well as advanced cancers and tumors. Both “tumor” and “cancer” as used herein refer to malignant (e.g., spreads from tissue of origin to another tissue) and non-malignant (e.g. grows but remains localized at the tissue of origin) e.g. benign growths.

[0044] Tumors of the central nervous system (also referred to herein as CNS tumors) are known in the art. See, e.g., D.N. Louis et al., The 2021 WHO Classification of Tumors of the064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Central Nervous System: a summary, Neuro Oncol. 2021 Aug; 23(8): 1231–1251. doi: 10.1093 / neuonc / noab106. Tumors of the central nervous system encompass brain and spinal cord tumors. CNS tumors include astrocytoma, oligodendroglioma, glioblastoma, diffuse astrocytoma, angiocentric glioma, polymorphous low-grade neuroepithelial tumor of the young, diffuse low-grade glioma, diffuse midline glioma, diffuse hemispheric glioma, diffuse pediatric-type high-grade glioma, infant-type hemispheric glioma, pilocytic astrocytoma, high- grade astrocytoma with piloid features, pleomorphic xanthoastrocytoma, subependymal giant cell astrocytoma, chordoid glioma, astroblastoma, ganglion cell tumors, dysembryoplastic neuroepithelial tumor, diffuse glioneuronal tumor with oligodendroglioma-like features and nuclear clusters, papillary glioneuronal tumor, rosette-forming glioneuronal tumor, myxoid glioneuronal tumor, diffuse leptomeningeal glioneuronal tumor, multinodular and vacuolating neuronal tumor, dysplastic cerebellar gangliocytoma (lhermitte-duclos disease), extraventricular neurocytoma, supratentorial ependymomas, posterior fossa ependymomas, spinal ependymomas, medulloblastoma, atypical teratoid / rhabdoid tumor, embryonal tumor with multilayered rosettes, CNS neuroblastoma, CNS tumor with bcor internal tandem duplication, desmoplastic myxoid tumor of the pineal region, meningiomas, solitary fibrous tumor, meningeal melanocytic tumors, adamantinoma Tous craniopharyngioma, and papillary craniopharyngioma.

[0045] A brain tumor is a cancerous (malignant) or non-cancerous (benign) abnormal growth of cells or mass in the brain.. Brain tumors can happen in the brain tissue. Brain tumors also can happen near the brain tissue. Nearby locations include nerves, the pituitary gland, the pineal gland, and the membranes that cover the surface of the brain. Brain tumors can begin in the brain. These are called primary brain tumors. Sometimes, cancer spreads to the brain from other parts of the body. These tumors are secondary brain tumors, also called metastatic brain tumors. Many different types of primary brain tumors exist. Some brain tumors are not cancerous. These are called noncancerous brain tumors or benign brain tumors. Noncancerous brain tumors may grow over time and press on the brain tissue. Other brain tumors are brain cancers, also called malignant brain tumors. Brain cancers may grow quickly. The cancer cells can invade and destroy the brain tissue. Brain tumors are a subset of CNS tumors. Brain tumors include but are not limited to: glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, and acoustic neuroma064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS (vestibular schwannoma). Some brain tumors are high grade (fast growing). Some brain tumors are low grade (slow growing).

[0046] Specifically, the PARP1 inhibitors disclosed herein are the compounds represented by Formula (I): (I) or a hydrate, or an acceptable salt thereof,wherein: R1 is selected from the group of halo, alkyl and haloalkyl; R2 is selected from the group of halo and alkyl; R3is selected from the group of halo, alkyl and cyano (that is —CN); R4is selected from the group of alkyl, haloalkyl and cycloalkyl.

[0047] In one or more embodiments of the compound of Formula (I), R1is halo, C1-3alkyl or halo C1-3 alkyl. In some embodiments, R1 is F, methyl or trifluoromethyl.

[0048] In one or more embodiments of the compound of Formula (I), R2is halo or C1-3alkyl. In some embodiments, R2is F, Cl or methyl.

[0049] In one or more embodiments of the compound of Formula (I), R3 is halo, C1-3 alkyl or cyano. In some embodiments, R3is F, methyl or cyano.

[0050] In one or more embodiments of the compound of Formula (I), R4is C1-3alkyl, halo C1-3 alkyl or C3-4 cycloalkyl. In some embodiments, R4 is methyl, ethyl, difluoroethyl or cyclopropyl.

[0051] In one or more embodiments of the compound of Formula (I), R1 is methyl; R2 is F; R3is F or cyano; R4is methyl or cyclopropyl.

[0052] In one or more embodiments of the compound of Formula (I), R1is F; R2is F, Cl or methyl; R3 is F or methyl; R4 is methyl, ethyl, difluoroethyl or cyclopropyl.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0053] In some embodiments of the compound of Formula (I), one or more of R1, R2, R3and R4 are alkyl, haloalkyl or cycloalkyl and are not further substituted. In some embodiments of the compound of Formula (I), one or more of R1, R2, R3and R4are alkyl, haloalkyl or cycloalkyl as defined herein and only R4 is further substituted e.g. is substituted with F, Cl, Br, I, —OH, —CHO, —NH2, or combinations thereof.

[0054] Exemplary compounds of Formula (I) include, without limitation: 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6-fluoro-3- methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 1); 7-((4-(2-cyano-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6-fluoro-3- methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 2); 7-((4-(2-fluoro-6-(cyclopropylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6- fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 3); 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3- trifluoromethyl-6-fluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 4); 7-((4-(2-methyl-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6- difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 5); 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6- difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 6); 7-((4-(2-fluoro-6-(ethylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6- difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 7); 7-((4-(2-fluoro-6-(2,2-difluoroethylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)- 3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 8); 7-((4-(2-fluoro-6-(cyclopropylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6- difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 9); 3-fluoro-7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6- chloropyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 10); 3-fluoro-7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6- methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (Example 11); or a hydrate, an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0055] Some of the compounds of the present disclosure may exist as stereoisomers including optical isomers. The disclosure includes all stereoisomers and the racemic mixtures of such stereoisomers as well as the individual enantiomers that may be separated according to methods that are well known to those of ordinary skill in the art.

[0056] Examples of pharmaceutically acceptable salts include inorganic and organic acid salts, such as hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate and oxalate; and inorganic and organic base salts formed with bases, such as sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) and N-methyl-glucamine.

[0057] Examples of prodrugs of the compounds of the disclosure include the simple esters of carboxylic acid-containing compounds (e.g., those obtained by condensation with a C1-4 alcohol according to methods known in the art); esters of hydroxy containing compounds (e.g., those obtained by condensation with a C1-4 carboxylic acid, C3-6 diacid or anhydride thereof, such as succinic anhydride and fumaric anhydride according to methods known in the art); imines of amino containing compounds (e.g., those obtained by condensation with a C1-4 aldehyde or ketone according to methods known in the art); carbamate of amino containing compounds, such as those described by Leu, et al., (J. Med. Chem.42: 3623-3628 (1999)) and Greenwald, et al., (J. Med. Chem. 42: 3657-3667 (1999)); and acetals and ketals of alcohol- containing compounds (e.g., those obtained by condensation with chloromethyl methyl ether or chloromethyl ethyl ether according to methods known in the art).

[0058] The PARP1 inhibitors of the present disclosure may be prepared using method known to those skilled in the art, or the novel methods of this disclosure. Specifically, the compounds of the disclosure with Formula (I) can be prepared as illustrated by the exemplary reaction in Scheme 1. Reaction of 4-methyl-1H-pyrazole-5-carboxylic acid and SOCl2 produced 4- methyl-1H-pyrazole-5-carbonyl chloride. Reaction of 4-methyl-1H-pyrazole-5-carbonyl chloride and 3-bromo-2,6-difluoroaniline under alkaline condition (such as under the catalysis of NaH, NaHMDS or LiHMDS (lithium bis(trimethylsilyl)amide)) produced N-(3-bromo-2,6- difluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide. Ring closing of N-(3-bromo-2,6- difluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide under alkaline condition (such as under the catalysis of K2CO3) produced 7-bromo-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin- 4(5H)-one. Reaction of 7-bromo-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one and (Bu)3SnCH2OH under the catalysis of Pd catalyst (such as Xphos Pd G2) produced 6-fluoro-064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS 7-(hydroxymethyl)-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one. Reaction of 6-fluoro-7- (hydroxymethyl)-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one and HBr produced 7- (bromomethyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one. Reaction of 7- (bromomethyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one and 6-fluoro-N- methyl-5-(piperazin-1-yl)picolinamide under alkaline condition (such as under the catalysis of DIEA and KI) produced the target compound 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3- yl)piperazin-1-yl)methyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one. Scheme 1replacement of 6-fluoro-N-methyl-5-(piperazin-1-yl)picolinamide with 6-cyano-N-methyl-5- (piperazin-1-yl)picolinamide produced the target compound 7-((4-(2-cyano-6- (methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6-fluoro-3-methylpyrazolo[1,5- a]quinoxalin-4(5H)-one. Replacement of 4-methyl-1H-pyrazole-5-carboxylic acid with 4- fluoro-1H-pyrazole-5-carboxylic acid produced the target compound 7-((4-(2-fluoro-6- (methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6-difluoropyrazolo[1,5- a]quinoxalin-4(5H)-one. Replacement of 3-bromo-2,6-difluoroaniline with 3-bromo-2-chloro- 6-fluoroaniline produced the target compound 3-fluoro-7-((4-(2-fluoro-6- (methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6-chloropyrazolo[1,5-a]quinoxalin- 4(5H)-one.

[0060] The compounds of this disclosure can be prepared as illustrated by the exemplary reaction in Scheme 2. Reaction of 4-fluoro-1H-pyrazole-5-carboxylic acid and SOCl2produced 4-fluoro-1H-pyrazole-5-carbonyl chloride. Reaction of 4-fluoro-1H-pyrazole-5-carbonyl chloride and 3,5-dibromo-2,6-difluoroaniline under alkaline condition (such as under the catalysis of NaH, NaHMDS or LiHMDS) produced N-(3,5-dibromo-2,6-difluorophenyl)-4-064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS fluoro-1H-pyrazole-5-carboxamide. Ring closing of N-(3,5-dibromo-2,6-difluorophenyl)-4- fluoro-1H-pyrazole-5-carboxamide under alkaline condition (such as under the catalysis of K2CO3) produced 7,9-dibromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one. Selective dehalogenation of 7,9-dibromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one in the presence of sodium ascorbate, CuI and DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), produced 7-bromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one. Reaction of 7-bromo-3,6- difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one and (Bu)3SnCH2OH under the catalysis of Xphos Pd G2 (chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′- amino-1,1′-biphenyl)]palladium(II)) produced 3,6-difluoro-7-(hydroxymethyl)pyrazolo[1,5- a]quinoxalin-4(5H)-one. Reaction of 3,6-difluoro-7-(hydroxymethyl)pyrazolo[1,5- a]quinoxalin-4(5H)-one and HBr produced 7-(bromomethyl)-3,6-difluoropyrazolo[1,5- a]quinoxalin-4(5H)-one. Reaction of 7-(bromomethyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin- 4(5H)-one and 6-fluoro-N-methyl-5-(piperazin-1-yl)picolinamide under alkaline condition (such as under the catalysis of DIEA and KI) produced the target compound 7-((4-(2-fluoro-6- (methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6-difluoropyrazolo[1,5- a]quinoxalin-4(5H)-one. Scheme 2

[0061] Other related compounds can be prepared using similar methods. For example, replacement of 4-fluoro-1H-pyrazole-5-carboxylic acid with 4-methyl-1H-pyrazole-5- carboxylic acid can produce the target compound 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin- 3-yl)piperazin-1-yl)methyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one. Replacement of 6-fluoro-N-methyl-5-(piperazin-1-yl)picolinamide with N,6-dimethyl-5- (piperazin-1-yl)picolinamide can produce the target compound 7-((4-(2-methyl-6- (methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6-difluoropyrazolo[1,5- a]quinoxalin-4(5H)-one.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0062] Also included within the scope of the present disclosure are the pharmaceutically acceptable salts of the PARP1 inhibitor(s) of the present disclosure. Acid addition salts are formed by mixing a solution of the compound(s) of the present disclosure with a solution of a pharmaceutically acceptable non-toxic acid, such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, and the like. Base addition salts are formed by mixing a solution of the compounds of the present disclosure with a solution of a pharmaceutically acceptable non-toxic base, such as sodium hydroxide, potassium hydroxide, hydrocholine, sodium carbonate, tris(hydroxymethyl)aminomethane, N-methyl-glucosamine and the like.

[0063] The compound(s) of the disclosure may be administered as a raw chemical. The compounds of the disclosure may also be administered as part of a suitable pharmaceutical preparation containing pharmaceutically acceptable carriers (comprising excipients and auxiliaries). Such pharmaceutically acceptable carriers facilitate the manufacture of pharmaceutically acceptable preparations from the compound(s). In some embodiments, the pharmaceutical preparations, for example, oral or other preparations such as tablets, dragees, and capsules, as well as solutions suitable for injection or oral administration, contain from approximately 0.01% w / w to 99% w / w, for example from approximately 0.25% w / w to 75% w / w of active compound(s), together with excipient(s).

[0064] The PARP1 inhibitor of the present disclosure can be administered in a pharmaceutical composition containing a pharmaceutically acceptable carrier, wherein the pharmaceutical composition includes all pharmaceutical preparations containing the compound of the present disclosure in an amount that can effectively achieve its intended goal. While individual needs vary, determination of the optimal amount of each part in the pharmaceutical preparation is within the skill of the art. Typically, the compounds or a hydrates, or an isotope- substituted derivative or the pharmaceutically acceptable salts thereof may be administered to mammals orally at a dose of about 0.0025 to 50 mg per kg body weight per day. In some embodiments, from approximately 0.01 mg / kg to approximately 10 mg / kg body weight is orally administered. If a known anticancer agent is also administered, it is administered in an amount that is effective to achieve its intended purpose. The optimal amounts of such known anticancer agents are well known to those skilled in the art.

[0065] The unit oral dose may comprise from approximately 0.01 to approximately 50 mg, for example approximately 0.1 to approximately 10 mg of the compound of the disclosure. The064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS unit dose may be administered one or more times, with one or more tablets daily, each containing from approximately 0.1 to 50 mg, conveniently approximately 0.25 to 10 mg of the compound of the disclosure or solvates thereof.

[0066] In topical preparations, the compound(s) of the disclosure may be present at a concentration of approximately 0.01 to 100 mg per gram of carrier.

[0067] The present disclosure also provides compositions. In some embodiments, the compositions of the invention are formulated using one or more pharmaceutically acceptable excipients or carriers. Exemplary diluents, carriers and excipients for use in the compositions disclosed herein are known to the person of ordinary skill in the art and include but are not limited to those disclosed in, e.g., the Handbook of Pharmaceutical Excipients (Raymond C. Rowe et al. eds., 6th edition 2009), the entire contents of which are incorporated herein by reference. In addition, Exemplary compositions e.g. dosage forms for the compositions disclosed herein are known to the person of ordinary skill in the art and include but are not limited to those disclosed in, e.g., Loyd V. Allen, Jr. et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems (8th ed.2005), the entire contents of which are incorporated herein by reference.

[0068] In one embodiment, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of at least one compound of Formula (I) disclosed herein (e.g. a therapeutically effective amount of one compound of the present disclosure), or any pharmaceutically acceptable form thereof (e.g., a pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers which are useful include, but are not limited to, glycerol, water, saline, ethanol and other pharmaceutically acceptable salt solutions such as phosphates and salts of organic acids. Examples of these and other pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey), the entire disclosure of which is incorporated by reference herein for all purposes.

[0069] The pharmaceutical preparations of the disclosure may be administered to any mammal, so long as they may experience the therapeutic effects of the compound(s) of the disclosure. Foremost among such mammals are humans and veterinary animals, although the disclosure is not intended to be so limited.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0070] The pharmaceutical preparations of the present disclosure may be administered by any means that achieve their intended purpose. For example, administration may be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes. Alternatively, or concurrently, administration may be by the oral route. The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, frequency of treatment, and the nature of the effect desired.

[0071] The pharmaceutical preparations of the present disclosure can be manufactured in a known manner, e.g., by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes. Pharmaceutical preparations for oral use may be obtained by combining the active compounds with solid excipient(s), optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.

[0072] Suitable excipients include, for example, fillers, such as saccharides, e.g. lactose or sucrose, mannitol or sorbitol; cellulose preparations and / or calcium phosphates, e.g. tricalcium phosphate or calcium hydrogen phosphate; as well as binders, such as starch paste, including maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinyl pyrrolidone. If desired, disintegrating agents may be added, such as the above-mentioned starches and carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate. Auxiliaries are, above all, flow-regulating agents and lubricants, e.g., silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, and / or polyethylene glycol. Dragee cores are provided with suitable coatings which, if desired, are resistant to gastric juices. For this purpose, concentrated saccharide solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose solutions, such as cellulose acetate phthalate or hydroxypropylmethyl-cellulose phthalate, are used. Dyes or pigments may be added to the tablets or dragee core coatings, e.g., for identification or in order to characterize combinations of active compound doses.

[0073] Other pharmaceutical preparations, which may be used orally, include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS as glycerol or sorbitol. The push-fit capsules may contain the active compounds in the form of granules, which may be mixed with fillers, such as lactose; binders, such as starches; and / or lubricants, such as talc or magnesium stearate, and stabilizers. In soft capsules, the active compound(s) are, for example, dissolved or suspended in suitable liquids, such as fatty oils, or liquid paraffin, in which stabilizers may be added.

[0074] Suitable formulations for parenteral administration include aqueous solutions of the active compounds, e.g., aqueous solutions and alkaline solutions of water-soluble salts. In addition, appropriate oily injection suspensions of the active compounds may be administered. Suitable lipophilic solvents or vehicles include fatty oils, e.g., sesame oil, or synthetic fatty acid esters, e.g., ethyl oleate or triglycerides or polyethylene glycol-400, or cremophor, or cyclodextrins. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, e.g., sodium carboxymethyl cellulose, sorbitol, and / or dextran. Optionally, the suspension may also contain stabilizers.

[0075] In accordance with one aspect of the present disclosure, compounds of the disclosure are employed in topical and parenteral formulations and are used for the treatment of skin cancer.

[0076] The topical formulations of this disclosure can be formulated as oils, creams, lotions, ointments and the like by choice of appropriate carriers. Suitable carriers include vegetable or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohol (greater than C12). In some embodiments, carriers are used in which the active ingredient(s) are soluble. Emulsifiers, stabilizers, humectants and antioxidants may also be included, as well as agents imparting color or fragrance, if desired. Additionally, transdermal penetration enhancers may be employed in these topical formulations. Examples of such enhancers are found in U.S. Patent Nos.3,989,816 and 4,444,762.

[0077] Creams are, for example, formulated from a mixture of mineral oil, self-emulsifying beeswax and water in which mixture of the active ingredient, dissolved in a small amount of an oil, such as almond oil, is admixed. A typical example of such a cream is one which includes approximately 40 parts of water, approximately 20 parts of beeswax, approximately 40 parts of mineral oil and approximately one part of almond oil.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0078] Ointments may be formulated by mixing a solution of the active ingredient(s) in a vegetable oil, such as almond oil, with warm soft paraffin and allowing the mixture to cool. A typical example of such an ointment is one which includes approximately 30% by weight of almond oil and approximately 70% by weight of white soft paraffin.

[0079] The composition containing the PARP1 inhibitor can be used (e.g. in the methods and uses disclosed herein) in combination with at least one known anti-cancer drug or a pharmaceutically acceptable salt of an anti-cancer drug. In particular, the composition can be used in combination with other anti-cancer drugs related to the mechanism of DNA damage and repair, including HDAC inhibitors Volinota, Romididesin, Papiseta and Bailesta; and so on. And the composition can be used in combination with other anti-cancer drugs related to cell division checkpoints, including Chk1 / 2 inhibitors, CDK4 / 6 inhibitors such as Palbociclib, Wee1 inhibitors, ATM inhibitors, ATR inhibitors, DNA-PK inhibitors and so on. Other known targeted anti-cancer drugs which may be used for anti-cancer combination therapy include, but are not limited to, PRMT5 inhibitors, Polθ inhibitors, RAD51 inhibitors and so on. Other known anti-cancer drugs which may be used for anti-cancer combination therapy include, but are not limited to, alkylating agents, such as busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cis-platin, mitomycin C, bleomycin and carboplatin; topoisomerase I inhibitors, such as camptothecin, irinotecan and topotecan; topoisomerase II inhibitors, such as doxorubicin, epirubicin, aclacinomycin, mitoxantrone, elliptinium and etoposide; RNA / DNA antimetabolites, such as 5-azacytidine, gemcitabine, 5-fluorouracil and methotrexate; DNA antimetabolites, such as 5-fluoro-2'- deoxy-uridine, fludarabine, nelarabine, ara-C, thioguanine, pralatrexate, pemetrexed, hydroxyurea and thioguanine; antimitotic agent, such as colchicine, vinblastine, vincristine, vinorelbine, paclitaxel, ixabepilone, cabazitaxel and docetaxel; antibodies, such as mAb, panitumumab, necitumumab, nivolumab, pembrolizumab, ramucirumab, bevacizumab, pertuzumab, trastuzumab, cetuximab, obinutuzumab, ofatumumab, rituximab, alemtuzumab, ibritumomab, tositumomab, brentuximab, daratumumab, elotuzumab, T-DM1, Ofatumumab, Dinutuximab, Blinatumomab, ipilimumab, avastin, herceptin and mabthera; Antibody–Drug Conjugates (ADC) such as T-DM1, Trastuzumab Deruxtecan, Trastuzumab Emtansine, Datopotamab Deruxtecan, Gemtuzumab Ozogamicin, Brentuximab Vedotin, Inotuzumab Ozogamicin, Sacituzumab govitecan, Enfortumab Vedotin, Belantamab Mafodotin; kinase inhibitors, such as imatinib, gefitinib, erlotinib, osimertinib, afatinib, ceritinib, alectinib, crizotinib, erlotinib, lapatinib, sorafenib, regorafenib, vemurafenib, dabrafenib, aflibercept,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS sunitinib, nilotinib, dasatinib, bosutinib, ponatinib, ibrutinib, cabozantinib, lenvatinib, vandetanib, trametinib, cobimetinib, axitinib, temsirolimus, Idelalisib, pazopanib, Torisel and everolimus. Other known anti-cancer drugs which may be used for anticancer combination therapy include tamoxifen, letrozole, fulvestrant, mitoguazone, octreotide, retinoic acid, arsenic trioxide, zoledronic acid, bortezomib, carfilzomib, Ixazomib, vismodegib, sonidegib, denosumab, thalidomide, lenalidomide, Venetoclax, Aldesleukin (recombinant human interleukin-2) and Sipueucel-T (prostate cancer treatment vaccine).

[0080] Further, in any embodiments of any of the methods and uses disclosed herein wherein a compound having the structure of Formula (I) or of any subset (e.g. subgenus) or species thereof disclosed herein, is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs or is used in combination with one or more additional anticancer therapies or drugs, the one or more additional anticancer therapies or drugs is selected from any such disclosed herein, for example, monoclonal antibodies, surgery, radiation therapy (such as ionizing radiation (IR), gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes), endocrine therapy, biologic response modifiers (such as interferons, interleukins, and tumor necrosis factor (TNF)), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, spindle poisons (such as vinblastine, vincristine, vinorelbine, and paclitaxel), podophyllotoxins (such as etoposide, irinotecan, and vopotecan), nitrosoureas (such as varmustine and lomustine), inorganic ions (such as cisplatin and carboplatin), enzymes (such as asparaginase), hormones (such as tamoxifen, leuprolide, flutamide, and megestrol), Gleevec™, adriamycin, dexamethasone, and cyclophosphamide.

[0081] The present disclosure also provides use of PARP1 inhibitors in the preparation of a medicament for the treatment of a central nervous system tumor, for example a brain tumor, in individuals. The central nervous system tumor may be selected from any of those disclosed herein. The brain tumor may be selected from any of those disclosed herein. In some embodiments, the PARP1 inhibitor is a compound of Formula (I), or a hydrate, or an isotope- substituted derivative, or a pharmaceutically acceptable salt thereof as described herein. The present disclosure also provides PARP1 inhibitors or pharmaceutical compositions or pharmaceutical preparations containing one or more than one (e.g. two, three, four or five) PARP1 inhibitors in a method for treating a central nervous system tumor, for example a brain064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS tumor, in a subject as disclosed herein, e.g. in a human or in a veterinary or zoo animal. In some embodiments, the PARP1 inhibitor is a compound of Formula (I), or a hydrate, or an isotope- substituted derivative, or a pharmaceutically acceptable salt thereof as described herein.

[0082] The present disclosure also provides the use of a PARP1 inhibitor as described herein in the manufacture of a medicament for treating a central nervous system tumor, for example a brain tumor, in a subject in need thereof. The present disclosure also provides the use of a PARP1 inhibitor or a pharmaceutical composition containing the same in a method for treating a central nervous system tumor, for example a brain tumor, in a subject in need thereof. The present disclosure further provides a method for treating a central nervous system tumor, for example a brain tumor, in a subject in need thereof, comprising administering to the subject an effective amount of a PARP1 inhibitor or a pharmaceutical composition comprising the same. In some embodiments, the PARP1 inhibitor is a compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof as described herein.

[0083] In a further aspect, the present disclosure provides a method for inducing regression of a central nervous system tumor in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, tumor regression is induced. In some embodiments, inducing regression of the tumor is evaluated from 7 to 84 days after administering the compound e.g. any intervening integer number of days after administering the compound, e.g. 8 days after, 9 days after, etc. or within any range between any two such values. In some embodiments, tumor regression is between 1% and 100%, between 5% and 100%, between 10% and 100%, between 25% and 100%, between 50% and 100%, between 75% and 100%, or is any percentage value between 1% and 100%, e.g. 2%, 3%, 4%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% etc. or within any range between any two such values. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments, the CNS tumor, for example, a brain tumor, has a volume at time t that is smaller than the064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS tumor’s volume on the first day of administering the compound and time t is any of day 7, day 84, or any day between day 7 and day 84. In some embodiments of the method for inducing regression of a central nervous system tumor in a subject, the central nervous system tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumour. In some embodiments of the method for inducing regression of a central nervous system tumor in a subject, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments of the method for inducing regression of a central nervous system tumor in a subject, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0084] In an aspect, the present disclosure provides a method for inducing increased regression of a central nervous system tumor in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS after administering the compound, increased tumor regression is induced. In some embodiments, inducing increased regression of the tumor is evaluated from 7 to 84 days after administering the compound e.g. any intervening integer number of days after administering the compound, e.g. 8 days after, 9 days after, etc. or within any range between any two such values. In some embodiments, tumor regression is between 1% and 100% e.g. between 5% and 100% e.g. any integer percentage value between 1% and 100%, e.g.2%, 3% etc. or within any range between any two such values. In some embodiments, the subject is a human or is a non- human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments of the method for inducing increased regression of a central nervous system tumor in a subject, the central nervous system tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumor. In some embodiments of the method for inducing increased regression of a central nervous system tumor in a subject, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments of the method for inducing increased regression of a central nervous system tumor in a subject, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope- substituted derivative, or a pharmaceutically acceptable salt thereof and one or more064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0085] In an aspect, the present disclosure provides a method for inhibiting growth of a central nervous system tumor in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, tumor growth is inhibited. In some embodiments, inhibiting growth of a central nervous system tumor is evaluated from 7 to 84 days after administering the compound, e.g. any intervening integer number of days after administering the compound, e.g. 8 days after, 9 days after, etc. or within any range between any two such values. In some embodiments, tumor growth is inhibited by 10% to 150% e.g. by 30% to 150% or 50% to 120% e.g. any integer percentage value between 10% and 150%,, e.g. by 30%, 40%, 50%, 60%, 70%, 100%, 125%, 150% etc. or within any range between any two such values. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments of the method for inhibiting growth of a central nervous system tumor in a subject, the central nervous system tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumor. In some embodiments of the method for inhibiting growth of a central nervous system tumor in a subject, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments of the method for inhibiting growth of a central nervous system tumor in a subject, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0086] In an aspect, the present disclosure provides a method for inhibiting PARP1 activity e.g. selectively inhibiting PARP1 activity relative to PARP2 activity in a subject comprising administering to the subject an effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, after administering the compound, inhibition of PARP1 activity is greater than inhibition of PARP2 activity in the subject. In some embodiments, the ratio of PARP1 activity inhibition to PARP2 activity inhibition in the subject is at least 50, at least 750, at least 1500, at least 2500, at least 5,000, at least 7,500, at least 10,000, at least 15,000, at least 20,000, at least 25,000, or within a range between any two of these numbers. In some embodiments, the ratio of PARP1 activity inhibition to PARP2 activity inhibition in the subject is between 50 and 2,000, between 750 and 15,000, between 2500 and 15,000, or between 12,500 and 25,000 e.g. between 15,000 and 25,000. In some embodiments, the subject is a human or is a non- human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, an effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope- substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0087] In an aspect, the present disclosure provides a method for treating a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), , or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject has a disease or condition that is characterized by or due to excessive cell death. In some embodiments, the disease or condition is stroke or a neurodegenerative disease.

[0088] In a further aspect, the present disclosure provides a method for treating cancer in a subject comprising administering to the subject a therapeutically effective amount of a compound having the structure of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is an ovarian, breast, pancreatic or prostate cancer. In some embodiments, the cancer is deficient in DNA damage repair e.g. the cancer is deficient in homologous recombination. In some embodiments, the subject has a central nervous system (CNS) tumor. In some embodiments, the subject has a brain tumor. In some embodiments, the subject is a human or is a non-human animal e.g. a veterinary animal or zoo animal e.g. as disclosed herein. In some embodiments, the compound is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs e.g. an anticancer therapy or drug as disclosed herein. In some embodiments, the subject has a central nervous system (CNS) tumor. In some embodiments, the CNS tumor is selected from a meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumour. In some embodiments, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In some embodiments, the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients. In embodiments, a therapeutically effective amount of the compound e.g. the compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, or of the pharmaceutically acceptable composition comprising the same, is administered.

[0089] In an aspect, the present disclosure provides the use of a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for the preparation of a medicament. In an aspect, the present disclosure provides the use of a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for the preparation of a medicament for use in treating any condition e.g. a cancer, such as any CNS tumor, for example, a brain tumor, as disclosed or described herein.

[0090] In an aspect, the present disclosure provides the use of a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for treating a condition disclosed herein or for a purpose disclosed herein.

[0091] In an aspect, the present disclosure provides a compound (e.g. an effective amount of the compound) having the structure of Formula (I) as disclosed herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof for use in a method of treating a condition disclosed herein or for use in a purpose disclosed herein.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0092] In an aspect, the present disclosure provides a method of treating a central nervous system (CNS) disease or condition e.g. a CNS tumor, for example a brain tumor, comprising administering a compound of Formula I as disclosed herein, wherein the R groups are defined as set forth herein, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, wherein the substituted tricyclic compound of Formula (I) can penetrate the blood brain barrier (BBB). In some embodiments, the substituted tricyclic compound of Formula (I) penetrates the blood brain barrier (BBB). In some embodiments, the CNS disease or condition is a brain tumor. In some embodiments, the central nervous system (CNS) disease or condition is a CNS tumor. In some embodiments, the central nervous system (CNS) disease or condition is a CNS tumor selected from meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma, glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumor. In some embodiments, the central nervous system tumor is any of the central nervous system tumors disclosed herein. In some embodiments, the CNS tumor is a brain tumor e.g. a brain tumor as disclosed herein. In some embodiments, the brain tumor is a high grade (fast growing) tumor. In some embodiments, the brain tumor is a low grade (slow growing) tumor. In some embodiments, the brain tumor is a glioma, embryonal brain tumor (e.g. medulloblastoma), ependymomas, glioblastoma, primary central nervous system lymphoma, pineal region tumor (e.g. germ cell tumor or pineal cell tumor), pituitary tumor, meningioma, or acoustic neuroma (vestibular schwannoma). In embodiments, a therapeutically effective amount of the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered. In some embodiments, the therapeutically effective amount of the compound or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof is administered in a pharmaceutically acceptable composition, wherein the pharmaceutically acceptable composition comprises the compound, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable diluents, carriers or excipients.

[0093] In an aspect, the present disclosure provides a pharmaceutically acceptable composition comprising a compound having the structure of Formula (I), or a hydrate, or an064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable diluent, carrier or excipient.

[0094] In some embodiments of the methods disclosed herein, the method comprises administering a therapeutically effective amount of a compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof as described herein. In some embodiments of the methods disclosed herein, the method comprises administering a therapeutically effective amount of a compound of Formula (I), or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, as described herein. In various embodiments, the pharmaceutically acceptable salt is an inorganic or organic acid salt, e.g. a hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate or oxalate salt. In various embodiments, the pharmaceutically acceptable salt is an inorganic or organic base salts formed with bases, e.g. a sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) or N-methyl-glucamine salt.

[0095] In any embodiments of any of the methods and uses disclosed herein wherein a compound having the structure of Formula (I) or of any subset (e.g. subgenus) or species thereof disclosed herein, is administered before, simultaneously with, or after the administration of one or more additional anticancer therapies or drugs or is used in combination with one or more additional anticancer therapies or drugs, the one or more additional anticancer therapies or drugs is selected from, for example, monoclonal antibodies, surgery, radiation therapy (such as ionizing radiation (IR), gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes), endocrine therapy, biologic response modifiers (such as interferons, interleukins, and tumor necrosis factor (TNF)), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, spindle poisons (such as vinblastine, vincristine, vinorelbine, and paclitaxel), podophyllotoxins (such as etoposide, irinotecan, and vopotecan), nitrosoureas (such as varmustine and lomustine), inorganic ions (such as cisplatin and carboplatin), enzymes (such as asparaginase), hormones (such as tamoxifen, leuprolide, flutamide, and megestrol), Gleevec™, adriamycin, dexamethasone, and cyclophosphamide.

[0096] In any of the methods of treatment disclosed herein, in some embodiments, the subject is a subject in need of such treatment. In various embodiments, the subject is a human. In some064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS embodiments, the subject is a non-human such as a veterinary animal (e.g. pig, cow, sheep, horse, dog, and cat) or a zoo or laboratory animal (e.g. any non-human primate or mammal).

[0097] In any of the uses of a compound disclosed herein in the treatment of a condition in a subject, in some embodiments, the subject is a subject in need of such treatment. In various embodiments, the subject is a human. In some embodiments, the subject is a non-human such as a veterinary animal (e.g. pig, cow, sheep, horse, dog, and cat) or a zoo or laboratory animal (e.g. any non-human primate or mammal).

[0098] In any of the uses of a compound disclosed herein for the preparation of a medicament for use in the treatment of a subject or for use in the treatment of a condition in a subject, in some embodiments, the subject is a subject in need of such treatment. In various embodiments, the subject is a human. In some embodiments, the subject is a non-human such as a veterinary animal (e.g. pig, cow, sheep, horse, dog, and cat) or a zoo or laboratory animal (e.g. any non- human primate or mammal).

[0099] The following examples are illustrative, but not limiting, of the method and compositions of the present disclosure. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the disclosure. EXAMPLES

[0100] All reagents were of commercial quality. Solvents were dried and purified by standard methods. Mass spectrum analyses were recorded on a quadrupole mass spectrometer fitted with an electrospray interface.1H NMR spectra was recorded at 400 MHz. Chemical shifts were recorded in parts per million (ppm) downfield from TMS (0.00 ppm), and J coupling constants were reported in hertz (Hz). Example 1 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-6-fluoro-3- methylpyrazolo[1,5-a]quinoxalin-4(5H)-one

[0101] a) Preparation of 4-methyl-1H-pyrazole-5-carbonyl chloride: A solution of 4-methyl- 1H-pyrazole-5-carboxylic acid (2.0 g, 15.9 mmol) in SOCl2 (20 mL) was stirred at 80oC for064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS 16 hours. After the reaction was completed, the solvent was removed under reduced pressure to afford the target product (2.2 g, yield: 91.7%) as an off-white solid.

[0102] b) Preparation of N-(3-bromo-2,6-difluorophenyl)-4-methyl-1H-pyrazole-5- carboxamide: To a solution of 3-bromo-2,6-difluoroaniline (2.5 g, 16.7 mmol) in THF (20 mL) was added NaH (60% in mineral oil, 2.776 g, 69.4 mmol) at 0oC. The mixture was stirred at 0oC under N2atmosphere for 0.5 hour, and then 4-methyl-1H-pyrazole-5-carbonyl chloride (2.0 g, 13.9 mmol) was added at 0oC. The mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction was poured into ice water (100 mL) slowly, extracted with EA (200 mL×3). The organic phase was washed with brine (100 mL×2), dried over Na2SO4and concentrated under reduced pressure. The residue was purified by column over silica gel (PE:EA=10:1) to afford the target product (2.7 g, yield: 67.0%) as a grey solid. MS(ESI): 315.90 [M+H]+.

[0103] c) Preparation of 7-bromo-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one: To a solution of N-(3-bromo-2,6-difluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (2.7 g, 8.6 mmol) in DMSO (27 mL) was added K2CO3 ( 2.4 g, 17.2 mmol). The mixture was stirred at 120oC overnight. After the reaction was completed, the reaction was poured into ice brine, the solid was collected by filtration to afford the target product (2.5 g, yield: 86.8%) as a white solid. MS(ESI): 293.95 [M-1]-.

[0104] d) Preparation of 6-fluoro-7-(hydroxymethyl)-3-methylpyrazolo[1,5-a]quinoxalin- 4(5H)-one: To a solution of 7-bromo-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (500.0 mg, 1.7 mmol) in dioxane (10 mL) was added (Bu)3SnCH2OH (816.3 mg, 2.55 mmol) and Xphos Pd G2 (133.2 mg, 0.2 mmol) at room temperature. The mixture was stirred at 90oC under N2atmosphere for 3 hours. After the reaction was completed, an aqueous of KF (1 M, 20 mL) was added at room temperature, the mixture was stirred at the same temperature for 10 minutes and filtered, and the filtrate was extracted with EA (500 mL × 3). The combined organic phase was washed with brine (100 mL × 2), dried over Na2SO4 and concentrated under reduced pressure. The residue was triturated with a mixture of PE and EA (V / V=1:1), and the solid was collected by filtration and dried to afford the target product (500 mg, crude) as a grey solid. MS(ESI): 248.05 [M+1]+, 246.05 [M-1]-.

[0105] e) Preparation of 7-(bromomethyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin- 4(5H)-one: To a solution of 6-fluoro-7-(hydroxymethyl)-3-methylpyrazolo[1,5-a]quinoxalin-064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS 4(5H)-one (500.0 mg, 2.0 mmol) in HBr (48% in H2O, 5 mL) was stirred at 80oC for 1 hour. After the reaction was completed, the reaction was concentrated under reduced pressure to afford the target product (500 mg, crude, yield: 79.9%) as a white solid. MS(ESI): 310.90 [M+1]+, 308.85 [M-1]-.

[0106] f) Preparation of 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1- yl)methyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one: To a solution of 7- (bromomethyl)-6-fluoro-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (200.0 mg, 0.67 mmol, 1.0 eq), KI (9.1 mg, 0.07 mmol) and 6-fluoro-N-methyl-5-(piperazin-1-yl)picolinamide (182.2 mg, 0.8 mmol) in acetonitrile (5 mL) was added N,N-diisopropyl ethyl amine (DIEA, 332.1 mg, 2.0 mmol) at room temperature. The resulting solution was stirred at 80oC for 1 hour. After the reaction was completed, the solvent was removed under reduced pressure. The residue was purified by Prep-TLC (DCM:MeOH=10:1) to give the title compound (19.8 mg, yield: 6.6% ) as a white solid. Example Compound MWLC-MS(ESI)1H NMR (400 MHz) DMSO-d6: δ 8.36 (d, J = 4.9 Hz, 1H), 7.89 (s, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 8.1 1 467.48468.20Hz, 1H), 7.55 – 7.49 (m, 1H), 7.30 – 7.24 (m, [M+H]+1H), 3.63 (s, 2H), 3.18 – 3.06 (m, 4H), 2.72 (d, J = 4.8 Hz, 3H), 2.60 – 2.51 (m, 4H), 2.40 (s, 3H). Examples 2-5

[0107] The following compounds of Examples 2-5 were prepared using a synthesis method similar to that described in Example 1. Example Compound MWLC-MS(ESI)1H NMR (400 MHz) DMSO-d6: δ 11.76 (s, 1H), 8.59-8.52 (m, 1H), 8.07 (d, J = 8.8 Hz, 1H), 7.90 (s, 1H), 7.84 (d, 2 474.5475.15+J = 8.4 Hz, 1H), 7.70 (d, J = 8.9 Hz, 1H), 7.27 [M+H](t, J = 7.7 Hz, 1H), 3.66 (s, 2H), 3.29-3.19 (m, 4H), 2.74 (d, J = 4.8 Hz, 3H), 2.65 – 2.53 (m, 4H), 2.40 (s, 3H). DMSO-d6: δ 11.75 (br, 1H), δ 8.33 (d, J = 5.2 O HN Hz, 1H), 7.91 (s, 1H), 7.85 (d, J = 8.0 Hz, F N 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.53 (dd, J = 3 NH O 493.52494.208.0 ,8.0 Hz, 1H), 7 N[M+H]+.28 (t, J = 6.8 Hz, 1H), 3.65 N F N (s, 2H), 3.14-3.11 (m, 4H), 2.83-2.82 (m, 1H), N 2.59-2.57 (m, 4H), 2.42 (s, 3H), 0.64-0.62 (m, 4H).064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS DMSO-d6: δ 8.50 (s, 1H),8.40-8.33 (m, 1H), 7.97 (d, J = 8.6 Hz, 1H), 7.79 (d, J = 8.0 Hz, 4 521.45522.351H), 7.52 (dd, J = 10.6, 8.0 Hz, 1H), 7.36 (dd, [M+H]+J = 8.6, 6.7 Hz, 1H), 3.67 (s, 2H), 3.15 – 3.12 (m, 4H), 2.72 (d, J = 4.8 Hz, 3H), 2.59-2.54 (m, 4H). DMSO-d6 HN : δ 11.98 (s, 1H), 8.45 – 8.38 (m, F O N 1H), 8.22 (d, J = 3.8 Hz, 1H), 7.91 (d, J = 8.2 NH O 5 467468.10Hz, 1H), 7.78 (d, J = 8.5 Hz, 1H), 7.47 (d, J = N N F .48 N[M+H]+8.2 Hz, 1H), 7.35 (t, J = 8.1 Hz, 1H), 3.70 (s, N 2H), 2.99 – 2.89 (m, 4H), 2.79 (d, J = 4.1 Hz, 3H), 2.66 – 2.56 (m, 4H), 2.46 (s, 3H). Example 6 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1-yl)methyl)-3,6- difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one

[0108] a) Preparation of 4-fluoro-1H-pyrazole-5-carbonyl chloride: The solution of 4- fluoro-1H-pyrazole-5-carboxylic acid (30 g, 230 mmol) in SOCl2 (300 mL) was stirred at 80oC for 2 hours. The mixture was concentrated under reduced pressure to give the target product (22.6 g, yield: 66%) as a white solid.

[0109] b) Preparation of N-(3,5-dibromo-2,6-difluorophenyl)-4-fluoro-1H-pyrazole-5- carboxamide: To a solution of 3,5-dibromo-2,6-difluoroaniline (81.2 g, 283.0 mmol,) in THF (100 mL) was added sodium bis(trimethylsilyl)amide (NaHMDS, 2 M in THF, 295.0 mL, 589.6 mmol) at 0oC under nitrogen. The mixture was stirred at 0oC for 30 minutes.4-Fluoro- 1H-pyrazole-5-carbonyl chloride (35.0 g, 235.9 mmol) was added slowly under nitrogen. The reaction mixture was stirred at 0oC for 30 minutes and warmed to room temperature and stirred for 1.5 hours. The mixture was quenched with water (500 mL) and extracted with EA (500 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product, which was washed with PE (1200 mL) for 2 times and washed with DCM (1200 mL) for 2 times to afford the target compound (105.0 g, yield: 93%) as a yellow solid. MS(ESI): 399.80 [M+H]+.

[0110] c) Preparation of 7,9-dibromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one: To a solution of N-(3,5-dibromo-2,6-difluorophenyl)-4-fluoro-1H-pyrazole-5-carboxamide (60.15 g, 150.8 mmol) in DMSO (600 mL) was added K2CO3(83.2 g, 603.1 mmol) under nitrogen. The mixture was stirred at 100oC overnight. The mixture was cooled to room temperature. Water (600 mL) was added, the resulting mixture was filtered, the solid was064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS washed with water (100 mL ^ 3). The residue was dried under reduced pressure, to give the target product (56.0 g, yield: 98%) as a yellow solid. MS(ESI): 377.75 [M-H]-.

[0111] d) Preparation of 7-bromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one: To a solution of 7,9-dibromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (44.0 g, 116.1 mmol) in DMF (420 mL) and H2O (70 mL) was added sodium ascorbate (18.4 g, 92.9 mmol), CuI (8.8 g, 46.4 mmol), 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU, 17.7 g, 116.1 mmol) under nitrogen. The mixture was stirred at 120oC overnight. The mixture was cooled to room temperature. Water (600 mL) was added, the resulting mixture was filtered, the solid was washed by (DCM / MeOH=15 / 1, 200 mL) for 3 times and concentrated under reduced pressure. The crude product was extracted by EA (2000 mL×3), the organic phase was concentrated to give the target product (17 g, yield: 49%) as a yellow solid. MS(ESI): 299.90 [M+H]+.

[0112] e) Preparation of 3,6-difluoro-7-(hydroxymethyl)pyrazolo[1,5-a]quinoxalin- 4(5H)-one: To a solution of 7-bromo-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (22.22 g, 74.1 mmol) in dried dioxane (230 mL) was added Xphos Pd G2 (5.8 g, 7.4 mmol) and (tributylstannyl)methanol (47.6 g, 148.1 mmol) under nitrogen. The mixture was stirred at 80oC for 16 hours. The mixture was cooled to room temperature and KF aqueous solution (1 M, 250 mL) was added. The resulted mixture was filtered and the filtrate was extracted with EA (200 mL×3). The organic phase was concentrated under reduced pressure. The solid was washed with DMF(100 mL×3) and the DMF phase was concentrated under reduced pressure . The combined crude product was washed by the mixed solvent of PE and EA (PE / EA=3 / 1, 100 mL×3) and the mixed solvent of DCM and MeOH (DCM / MeOH=15 / 1, 50 mL×2) . The residue was dried to give the target compound (14.7 g, yield: 79%) as a white solid. MS(ESI): 252.05[M+H]+.

[0113] f) Preparation of 7-(bromomethyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)- one: The solution of 3,6-difluoro-7-(hydroxymethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one (14.2 g, 56.5 mmol) in HBr (48% in water, 200 mL) was stirred at 80oC for overnight under nitrogen. The mixture was concentrated under reduced pressure to give the title product (16.6 g, yield: 93%) as a white solid. MS(ESI): 313.90 [M+H]+.

[0114] g) Preparation of 7-((4-(2-fluoro-6-(methylcarbamoyl)pyridin-3-yl)piperazin-1- yl)methyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one: To a solution of 7- (bromomethyl)-3,6-difluoropyrazolo[1,5-a]quinoxalin-4(5H)-one (16.6 g, 52.8 mmol) in064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS acetonitrile (200 mL) was added KI (875.9 mg, 5.3 mmol), 6-fluoro-N-methyl-5-(piperazin-1- yl)picolinamide hydrochloride (15.4 g, 52.8 mmol), and DIEA (61.3 g, 474.9 mmol) at room temperature under nitrogen. The mixture was stirred at 80oC for 4 hours. After the reaction was completed, the solvent was removed under reduced pressure. To the residue was added DMF and the mixture was stirred at 45oC for 15 minutes. The mixture was cooled to room temperature and filtered to give the title compound (8.3 g) as a white solid.ExampleCompoundMW LC-MS(ESI)1H NMR (400 MHz)DMSO-d6: δ 8.40 (q, J = 4.8 Hz, 1H), 8.22 (d, F O HN F N J = 3.7 Hz, 1H), 7.91 (d, J = 8.3 Hz, 1H), 7.83 NH O 6 (d, J = 7.0 Hz, 1H), 7.56 (dd, J = 10.4, 8. N N F471.44472.152 Hz, N[M+H]+1H), 7.34 (dd, J = 8.4, 7.1 Hz, 1H), 3.68 (s, N 2H), 3.21 – 3.11 (m, 4H), 2.76 (d, J = 4.8 Hz, 3H), 2.63 – 2.55 (m, 4H). Examples 7-11

[0115] The following compounds of Examples 7-11 were prepared using a synthesis method similar to that described in Example 1. Example Compound MWLC-MS(ESI)1H NMR (400 MHz) DMSO-d6: δ 8.47 (t, J = 5.8 Hz, 1H), 8.22 (d, J = 3.7 Hz, 1H), 7.91 (d, J = 8.5 Hz, 1H), 7.83 7 485.47486.35(d, J = 8.5 Hz, 1H), 7.56 (dd, J = 10.4, 8.1 Hz, [M+H]+1H), 7.34 (dd, J = 8.8, 7.2 Hz, 1H), 3.68 (s, 2H), 3.27 - 3.23 (m, 2H), 3.19 - 3.13 (s, 4H), 2.63 – 2.48 (s, 4H), 1.08 (t, J = 7.2 Hz, 3H). DMSO-d6: δ 8.74 (t, J = 6.1 Hz, 1H), 8.22 (d, J = 3.8 Hz, 1H), 7.91-7.86 (m, 2H), 7.61 – 7.52 8 521.45522.05(m, 1H), 7.38 – 7.30 (m, 1H), 6.10 (tt, J = 56.6, [M+H]+4.2 Hz, 1H), 3.68 (s, 2H), 3.62 (dt, J = 15.3, 5.1 Hz, 2H), 3.22-3.14 (m, 4H), 2.62-2.57 (m, 4H). DMSO-d6: δ8.38 (d, J = 4.8 Hz, 1H), 8.21 (d, J = 3.8 Hz, 1H), 7.90 (d, J = 8.8 Hz, 1H), 7.83 9497.48498.60(d, J = 7.6 Hz, 1H), 7.56 (dd, J = 10.4, 8.3 Hz, [M+H]+1H), 7.36 - 7.29 (m, 1H), 3.68 (s, 2H), 3.18 – 3.12 (m, 4H), 2.88 – 2.80 (m, 1H), 2.64 – 2.56 (m, 4H), 0.67 – 0.62 (m, 4H). DMSO-d6: δ 8.39 (q, J = 5.0 Hz, 1H), 8.18 (d, J = 3.8 Hz, 1H), 8.06 (d, J = 8.5 Hz, 1H), 10 487.90488.557.83 – 7.77 (m, 1H), 7.53 (dd, J = 10.7, 8.0 Hz, [M+H]+1H), 7.42 (d, J = 8.5 Hz, 1H), 3.68 (s, 2H), 3.20-3.08 (m, 4H), 2.72 (d, J = 4.8 Hz, 3H), 2.64-2.52 (m, 4H). DMSO-d6: δ 11.04 (s, 1H), 8.42 (q, J = 5.3 Hz, 1H), 8.18 (d, J = 3.7 Hz, 1H), 7.95 (d, J = 8.4 11 467.48468.05Hz, 1H), 7.83 (d, J = 7.9 Hz, 1H), 7.55 (dd, J = [M+H]+10.6, 8.0 Hz, 1H), 7.26 (d, J = 8.4 Hz, 1H), 3.59 (s, 2H), 3.20-3.08 (m, 4H), 2.75 (d, J = 4.7 Hz, 3H), 2.60-2.56 (m, 4H), 2.47 (s, 3H).064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Example 12 PARP1 and PARP2 chemiluminescent Assay

[0116] The solution of recombinant poly(ADP-ribose) polymerase 1 and 2 (PARP1 and PARP2) (40 ng enzyme / well) and the compounds to be tested were mixed, respectively. The solutions were added to a 96-well plate coated with histone mixture, incubated at room temperature for 1 h, then 50 μL 0.3ng / mL Streptavidin-HRP was added to each well. The plates were incubated for 30 minutes at room temperature. Finally, the plates were treated with streptavidin-HRP followed by addition of the ELISA ECL substrate to produce chemiluminescence that can be measured using a chemiluminescence reader. Inhibition of the tested compound to PARP1 / 2 enzyme activity was calculated according to the following formula. Readings of positive control − ^ ^^ℎ^^^^^^^^%^ = ^ !"^^#$ ^% &^$^^^' (^^^)^* − ^ !"^^#$ ^% ^ #^^^' (^^^)^* IC50value is obtained by fitting the s-shaped dose response curve equation by using XL Fit software. The curve equation is Y=100 / (1+10^(logC-logIC50)), C is the compound concentration.

[0117] Table 1 summarizes the inhibitory effects of compounds on PARP1 and PARP2 enzyme activity (IC50). Table 1 ExampleIC50 (nM) Ratio of PARP 1 inhibition toPARP1 PARP2 PARP2 inhibition1 1.38 >10000 >7246 2 0.48 2504 5217 3 0.48 731.1 1523 4 0.50 67.13 134 5 1.06 2821.75 2662 6 0.53 462.44 873 7 0.24 431 1796 8 0.25 587 2348 9 0.23 314.97 1369 10 0.98 >10000 >10204 11 0.25 188 752064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0118] Relative to the PARP2 enzyme, the compounds tested have potent and selective inhibitory effects on the PARP1 enzyme. Example 13 Growth inhibition assays against BRCA mutant human breast cancer MDA-MB-436 cell line

[0119] The cells were cultured in complete medium (DMEM medium +10% FBS+ Insulin + glutathione). When the confluence reached about 80%, cells were digested and gently dispensed from the bottom of the dish with a 1 mL pipette. Cell suspension was collected and centrifuged at 500rpm for 3min. The supernatant was discarded, and the cell pellet were re- suspended in complete medium. The cells were seeded into a culture dish at an appropriate proportion, and then cultured in a 5% CO2incubator at 37℃. The assay was carried out when the cells were in optimum condition and the confluence was reached 80%. Cells in the logarithmic growth phase were taken to centrifugate, and the culture supernatant was removed. The cells were resuspended in refresh complete medium and counted. The resuspended cells were seeded at 3000 / well in a 96-well plate and incubated at 37℃, 5% CO2incubator overnight. The compound was prepared as below: 1000× dilution tested compound solution to 40× test compound solution by adding 5μL 1000× compound solution to 120 μL Medium (25-fold dilution). The solution was mixed by oscillation.0.1% DMSO was used as the control.

[0120] The next day, the 96-well plate inoculated with cells was taken out from the incubator, and the culture supernatant was removed. Then fresh medium of 195 uL / well and 5μL / well of 40× test compound solution as mentioned above were added into the 96 well plate, respectively. Finally, the plate was incubated for 7 days in a 37℃ 5% CO2 incubator. The medium containing compound was changed on the fourth day. After 7 days, 20μL of CCK-8 was added to each well and shaken gently, then was cultured for 4 hours. The plate was shaken for 5min after incubation. the absorbance values of 450nm or 650nm wavelengths were recorded respectively (OD = absorbance value of 450nm - absorbance value of 650nm) by using the multifunction readout instrument.

[0121] Data were analyzed by software GraphPad Prism 6.0. The inhibitory activity of compounds on cell proliferation was plotted using cell survival rate against the compound concentration as coordinates. Cell survival rate %= (ODcompound- ODbackground) / (ODDMSO- ODbackground) ×100. The IC50 value was fitted by the s-shaped dose response curve equation: Y=100 / (1+10^(logC-logIC50)), and C was the compound concentration.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0122] Table 2 summarizes the inhibitory effect data (IC50) of the compounds on the proliferation of human breast cancer cells MDA-MB-436. Table 2 Example IC50 (nM) 1 2.99 2 1.58 3 4.50 4 4.49 5 1.72 6 1.32 7 1.24 8 0.62 9 1.90 10 5.58 11 4.98

[0123] The compounds tested have good inhibitory effect on the proliferation of BRCA mutated human breast cancer cells MDA-MB-436. Example 14 Pharmacokinetics studies of the compounds in plasma and brain following single oral administration to mice

[0124] The compounds of the present invention were formulated into a 0.5% methylcellulose / water uniform suspension, and were administered to CD-1 (ICR) mice by gavage at 10 mg / kg. The mice were euthanized at 0.5, 4 and 8 hours post-dose, respectively, using CO2inhalation and then blood and brain were collected from each mouse, respectively. Each blood collection (about 0.1 mL per time point) was performed from saphenous vein or other suitable site of each mouse into pre-chilled EDTA-K2 tubes. After collection, the brain was washed with cold saline, wiped dry and weighed. Brain was homogenized using homogenizing buffer (15 mM PBS (pH7.4):MeOH=2:1) at the ratio of 1:4 (1 g tissue with 4 mL buffer, the dilution ratio is 5). The tissue homogenate was kept at -60 °C or lower until LC- MS / MS analysis. Concentrations of the compound in plasma and brain were determined by the LC-MS / MS method.

[0125] The concentrations, calculated AUC0-8h, and B / P ratio of the tested compounds are summarized in Table 3.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Table 3 Example Time Concentration Concentration AUC0-8h in B / P point (h) in plasma in brain (ng / g) brain ratio (ng / mL) (h·ng / mL) * 0.5 4650 2087 0.47 1 4 3970 2705 16389 0.68 8 1793 1228 0.69 0.5 4097 1260 0.31 2 4 3817 900 6037 0.24 8 981 234 0.24 0.5 2313 1450 0.64 3 4 2173 1982 11359 0.91 8 872 722 0.83 0.5 18587 537 0.03 4 4 36100 1377 8541 0.04 8 38567 1158 0.03 0.5 18733 2128 0.12 5 4 26567 2252 14877 0.09 8 13967 1198 0.09 0.5 17002 1556 0.09 6 4 14265 1966 15031 0.14 8 10844 2273 0.21 0.5 4625 1864 0.40 7 4 4887 1758 12616 0.38 8 4910 1186 0.25 0.5 5909 1406 0.24 8 4 3685 943 7610 0.29 8 2983 673 0.26 0.5 1133 204 0.16 9 4 2593 455 3490 0.17 8 3897 688 0.18 0.5 4625 1864 0.40 10 4 4887 1758 12616 0.38 8 4910 1186 0.25 0.5 775 551 0.71 11 4 1046 666 4656 0.66 8 1313 533 0.44 Note: * Compound concentration versus time data was analyzed by non-compartmental approaches using the WinNonlin software program, and AUC0-8h was calculated using the linear logarithmic trapezoidal method.

[0126] The compounds tested have good exposure in brain in mice. Example 15 Kp,uu of the compounds in mice

[0127] The ratio of unbound drug in brain to unbound drug in plasma (Kp,uu) was determined as follows.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Determination of fraction unbound in mouse plasma

[0128] On the day of experiment, the mouse plasma was thawed under running cold tap water and centrifuged at 3220 ×g for 5 min to remove any clots. The pH value was checked and recorded. Only plasma within range of pH 7.0 to pH 8.0 was used.

[0129] The test compounds or positive control warfarin was spiked into frozen mouse plasma at the final concentration of 2 μM. An aliquot of 150 µL of the compound-spiked plasma sample was added to one side of the chamber in a 96-well equilibrium dialysis plate (HTD dialysis) and an equal volume of dialysis buffer was added to the other side of the chamber. An aliquot of plasma sample was harvested before the incubation and used as T0 samples for recovery calculation. Triplicate incubations were performed. The plate was then incubated in a humidified incubator with 5% CO2 at 37°C for 4 hours. After incubation, 50 µL samples were taken from the plasma side as well as the buffer side. The plasma sample was mixed with an equal volume of blank buffer; buffer samples were mixed with an equal volume of blank plasma. The matrix-matched samples were quenched with stop solution containing internal standard (IS). Samples were analyzed by LC-MS / MS. Test compound concentrations in plasma and buffer samples were determined based on peak area ratio of analyte to IS without a standard curve. Determination of fraction unbound in mouse brain homogenate

[0130] On the day of experiment, CD-1 mouse brain homogenate was thawed in water bath at room temperature and then incubated at 37 °C for 10 min before use. The test compound or control compound propranolol was spiked into blank brain homogenate at the final concentrations of 2 µM.

[0131] An aliquot of 100 µL of the compound-spiked brain homogenate sample was added to one side of the chamber in a 96-well equilibrium dialysis plate (HTD dialysis) and an equal volume of dialysis buffer (100 mM sodium phosphate and 150 mM NaCl, 7.4 ± 0.1) was added to the other side of the chamber. An aliquot of brain homogenate sample will be harvested before the incubation and used as T0 sample for recovery calculation. Triplicate incubations were performed.

[0132] The plate was then be rotated at approximately 100 rpm in a humidified incubator with 5% CO2 at 37°C for 4 hours. After incubation, 50 µL samples were taken from the brain064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS homogenate side as well as the buffer side. The brain homogenate sample was mixed with an equal volume of buffer; buffer samples were mixed with an equal volume of blank brain homogenate. The matrix-matched samples were quenched with stop solution containing internal standard (IS).

[0133] Samples were analyzed by LC-MS / MS. Test compound concentrations in brain homogenate and buffer samples were determined based on peak area ratio of analyte to IS without a standard curve.

[0134] The Kp,uuvalue was calculated by the formula: Kp,uu= (AUCbrain / AUCplasma)^(fu,brain / fu,plasma), wherein AUCbrain and AUCplasma are AUC0-8h in plasma and brain obtained from Example 14, respectively; fu,plasmaand fu,brainmean unbound (%) in plasma and brain, respectively. The Kp,uu value of the compounds in mice are summarized in Table 4. Table 4 Example fu,plasma fu,brain AUCplasma (h·ng / mL) AUCbrain (h·ng / mL) Kp,uu 1 2.0 2.2 27171 16389 0.663 5 0.5 3.8 162344 14877 0.699 6 1.1 4.3 108733 15031 0.540 10 1.2 1.8 37396 12616 0.505

[0135] The compounds tested have good Kp,uu. Example 16 Pharmacokinetic studies of the compounds following single oral administration to mice

[0136] Exemplary compounds of the present invention were formulated into a 0.5% methylcellulose / water uniform suspension, and were administered to CD-1 (ICR) mice by gavage at 10 mg / kg. Plasma samples were collected at 8 time points of 0.250, 0.500, 1.00, 2.00, 4.00, 6.00, 8.00 and 24.0 hours post-dose. Each blood collection (about 0.1 mL per time point) was performed from saphenous vein or other suitable site of each mouse into pre-chilled EDTA-K2 tubes. Concentrations of the compound were determined by LC-MS / MS method.

[0137] Plasma concentration versus time data was analyzed by non-compartmental approaches using the WinNonlin software program, and pharmacokinetic parameters were calculated using the linear logarithmic trapezoidal method. The mouse pharmacokinetic parameters of the compounds are summarized in Table 5, wherein, half-life t1 / 2means the time064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS it takes for half the drug concentration to be eliminated; Cmaxmeans the maximum plasma concentration; AUC0-t means the area under the plasma concentration-time curve from time 0 to the time of last; AUC0-infmeans the area under the plasma concentration-time curve from time 0 to infinity. Table 5 Example t1 / 2 (h) Cmax (ng / mL)AUC0-t (h·ng / mL) AUC0-inf (h·ng / mL) 1 3.43 4520 37212 37529 3 8.05 618 6593 7998 5 26.6 17867 311006 677057 6 8.80 25678 371983 446442 8 5.26 6530 46656 48717 10 9.45 4793 57428 70053

[0138] The compounds tested have good exposure in plasma in mice. Example 17 Anti-tumor efficacy studies in MDA-MB-436 human breast cancer subcutaneous model

[0139] Human breast cancer MDA-MB-436 cells were ordered from ATCC (HTB-130). The cells were cultured in L-15 medium with 10 µg / mL insulin, 16 µg / mL glutathione, 10% fetal bovine serum at 37 ºC in an atmosphere of 0 % CO2in air. The cells were routinely sub-cultured two or three times per week. The cells growing in an exponential growth phase were harvested and counted for tumor inoculation.

[0140] The cells (1×107cells in 0.2 mL medium with 50% Matrigel) were inoculated subcutaneously at the right flank in each mouse. The mice were randomized and treatment was started when the average tumor volume reached approximately 150 mm3for the efficacy study.

[0141] After grouping and drug treatment start, animal’s conditions were monitored every day. Body weight and tumor size were measured three times per week.

[0142] Animal body weight was monitored regularly as an indirect measurement of toxicity.

[0143] Tumor size was measured thrice weekly in two dimensions using a caliper, and the volume was expressed in mm3using the formula: V = 0.5a ^ b2, where a and b are the long and short diameters of the tumor, respectively.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS

[0144] The tumor size was then used for calculations of TGI (%) values. TGI (%)=[1-(Vtreat-t-Vtreat-1) / (Vcontrol-t-Vcontrol-1)] ^100, represents antitumor effectiveness, in which Vtreat-1 and Vcontrol-1are the mean volumes of the treated and control groups on grouping day (grouping day is the day the animals are randomized to groups, which is the first dosing day); Vtreat-t and Vcontrol-tare the mean volumes of the treated and control groups on a given day. When the TGI (%) value was greater than 100%, tumor regression (%) was calculated by the formula: (Vtreat-1-Vtreat-t) / Vtreat-1×100. Study-1

[0145] The anti-tumor efficacy of Compounds 1 and 6 (that is the compounds of Examples 1 and 6) in MDA-MB-468 human breast cancer subcutaneous xenograft tumor model was evaluated.

[0146] The grouping and dosing schedule are shown in Table 6. Table 6 Group Na CompoundDosage Dosing volume Route of Dosing therapy(mg / kg) (µL / g)cadministrationdregimene1 8 VehiclebNA 10 p.o QD ^28D 2 8 Compound 1 0.2 10 p.o QD ^ 28D 3 8 Compound 1 1 10 p.o QD^28D 4 8 Compound 6 0.2 10 p.o QD^28D 5 8 Compound 6 1 10 p.o QD^28D Note: a. N is the number of animals, 8 tumor-bearing nude mice per group; b. The vehicle is 0.5%MC in water; c.Dosing volume: adjust dosing volume based on body weight 10µL / g. Treatmentschedule may be adjusted if body weight loss >15%; d. p.o. is oral administration; e. QD is once a day; continuous administration for 28 days. NA means not applicable. Results: 1) Mortality, Morbidity, and Body Weight Gain or Loss Throughout the whole study, no obvious body weight loss was observed, and no death or any signs of morbidity were found. 2) Tumor growth inhibition and regression Tumor growth inhibition information including tumor volume and the TGI value compared to control group based on data of day 28 is shown in Table 7. Table 7 Tumor Volume on day Group Compound Therapy 28 (mm3)aTGI (%) Regression (%)064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS 1 Vehicle 1161.61±99.42 NA NA 2 Compound 1, 0.2 mg / kg 771.55±37.64 38.57 NA 3 Compound 1, 1 mg / kg 405.52±31.89 74.89 NA 4 Compound 6, 0.2 mg / kg 139.03±7.42 101.32 9 5 Compound 6, 1 mg / kg 92.95±8.76 105.89 39 Note: a. Mean ± SEM. NA means not applicable.

[0147] In summary, all treatment groups showed significant tumor growth inhibition compared with vehicle group. Compound 1 treatments showed good dose-dependent relationship at 0.2 mg / kg and 1 mg / kg. Moreover, Compound 6 treatments at 0.2 mg / kg and 1 mg / kg exhibited tumor regression.

[0148] During the study, no obvious body weight loss was found in the groups, animals behaved normally, and there was no death or any signs of morbidity, which indicated the compounds were well tolerated at the tested dose levels in mice. Study-2

[0149] The anti-tumor efficacy of Compounds 5 and 6 (that is the compounds of Examples 5 and 6) in MDA-MB-436 human breast cancer subcutaneous xenograft tumor model was evaluated.

[0150] The grouping and dosing schedule are shown in Table 8. Table 8 Group Na CompoundDosage Dosing volume Route of Dosing therapy(mg / kg) (µL / g)cadministrationdregimene1 8 VehiclebNA 10 p.o QD^28D 2 8 Compound 5 0.1 10 p.o QD^28D 3 8 Compound 6 0.1 10 p.o QD^28D 4 8 Compound 6 1 10 p.o QD^28D 5 8 Compound 6 10 10 p.o QD^28D Note: a. N is the number of animals, 8 tumor-bearing nude mice per group; b. The vehicle is 0.5%MC in water; c. Dosing volume: adjust dosing volume based on body weight 10µL / g. Treatment schedule may be adjusted if body weight loss >15%; d. p.o. is oral administration; e. QD is once a day; continuous administration for 28 days. NA means not applicable. Results: 1) Mortality, Morbidity, and Body Weight Gain or Loss064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Throughout the whole study, no obvious body weight loss was observed, and no death or any signs of morbidity were found. 2) Tumor growth inhibition and tumor regression

[0151] Tumor growth inhibition information including tumor volume and TGI value compared to control group based on data of day 28 was shown in Table 9. Table 9 Group Compound TherapyTumor Volume on dayRegression 28 (mm3) a TGI (%)(%) 1 Vehicle 948.08±90.41 NA NA 2 Compound 5, 0.1 mg / kg 256.12±15.74 86.64 NA 3 Compound 6, 0.1 mg / kg 188.2±19.47 95.20 NA 4 Compound 6, 1 mg / kg 120.71±6.81 103.67 19 5 Compound 6, 10 mg / kg 104.13±10.35 105.73 31 Note: a. Mean ± SEM. NA means not applicable.

[0152] In summary, all treatment groups showed significant tumor growth inhibition compared with vehicle group. Compounds 5 and 6 showed similar efficacy at 0.1 mg / kg. Moreover, Compound 6 treatments at 1 mg / kg and 10 mg / kg exhibited tumor regression.

[0153] During the study, no obvious body weight loss was found in the groups, animals behaved normally, and there was no death or any signs of morbidity, which indicated the compounds were well tolerated at the tested dose levels in mice. Example 18 Anti-tumor efficacy studies in DLD-1 BRCA2- / -human colorectal adenocarcinoma epithelial subcutaneous model

[0154] The anti-tumor efficacy of Compound 6 in DLD-1 BRCA2- / -human colorectal adenocarcinoma epithelial cancer subcutaneous xenograft tumor model was evaluated.

[0155] DLD-1 BRCA2- / -cells were constructed from DLD-1 wild type cells (purchased from ATCC), of which the BRCA2 gene was knocked out using CRISPR / Cas9 technology, by the laboratory of ICE Bioscience. DLD-1 BRCA2- / -cells were cultured in RPMI-1640 medium with 10% fetal bovine serum and 100 μg / mL Hygromycin B at 37 ºC in an atmosphere of 5 %064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS CO2in air. The cells growing in an exponential growth phase were harvested and counted for tumor inoculation.

[0156] The cells (5×106cells in 0.1 ml medium with 50% Matrigel) were inoculated subcutaneously at the right flank in each mouse. The mice were randomized and treatment was started when the average tumor volume reached approximately 175 mm3for the efficacy study. The grouping and dosing schedule are shown in Table 10. Table 10 Group Na CompoundDosage Dosing volume Route of Dosing therapy(mg / kg) (µL / g)cadministrationdregimene1 8 VehiclebNA 10 p.o QD^28D 2 8 Compound 6 0.2 10 p.o QD^28D 3 8 Compound 6 1 10 p.o QD^28D Note: a. N is the number of animals, 8 tumor-bearing nude mice per group; b. The vehicle is 0.5%MC in water; c. Dosing volume: adjust dosing volume based on body weight 10µL / g. Treatment schedule may be adjusted if body weight loss >15%; d. p.o. is oral administration; e. QD is once a day; continuous administration for 28 days. NA means not applicable.

[0157] After grouping and drug treatment start, animal’s conditions were monitored every day. Body weight and tumor size were measured twice per week.

[0158] Animal body weight was monitored regularly as an indirect measurement of toxicity.

[0159] Tumor size was measured thrice weekly in two dimensions using a caliper, and the volume was expressed in mm3using the formula: V = 0.5a ^ b2, where a and b are the long and short diameters of the tumor, respectively.

[0160] The tumor size was then used for calculations of TGI (%) values. TGI (%)=[1-(Vtreat-t-Vtreat-1) / (Vcontrol-t-Vcontrol-1)] ^100, represents antitumor effectiveness, in which Vtreat-1and Vcontrol-1 are the mean volumes of the treated and control groups on grouping day; Vtreat-t and Vcontrol-tare the mean volumes of the treated and control groups on a given day. When the TGI (%) value was greater than 100%, tumor regression (%) was calculated by the formula: (Vtreat-1-Vtreat-t) / Vtreat-1×100. Results: 1) Mortality, Morbidity, and Body Weight Gain or Loss064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Throughout the whole study, no obvious body weight loss was observed, and no death or any signs of morbidity were found. 2) Tumor growth inhibition

[0161] Tumor growth inhibition information including tumor volume and TGI value compared to control group based on data of day 28 is shown in Table 11. Table 11 Group Compound TherapyTumor Volume on dayRegression 28 (mm3) a TGI (%)(%) 1 Vehicle 1100.61±100.63 NA NA 3 Compound 6, 0.2 mg / kg 174.20±31.06 100.05 0.27 4 Compound 6, 1 mg / kg 49.23±6.23 113.43 72 Note: a. Mean ± SEM. NA means not applicable.

[0162] In summary, all treatment groups showed significant tumor growth inhibition compared with vehicle group, and Compound 6 showed good dose-dependent relationship at 0.2 mg / kg and 1 mg / kg.

[0163] During the study, no obvious body weight loss was found in the groups, animals behaved normally, and there was no death or any signs of morbidity, which indicated the compounds were well tolerated at current dose levels in mice. Example 19 Anti-tumor efficacy studies in MDA-MB-436-luc human breast cancer intracranial model

[0164] The MDA-MB-436-luc tumor cell line were maintained in vitro as monolayer culture in DMEM medium supplemented with 10% FBS and 1200 ng / mL puromycin at 37°C in an atmosphere of 5% CO2. The medium was renewed every 1 to 2 days. The tumor cells were routinely sub-cultured when reached about 80%-90% confluence by 0.25% trypsin-EDTA treatment, not to exceed 4-5 passages. The cells growing in an exponential growth phase were harvested and counted for tumor inoculation.

[0165] Animals were anesthetized by s.c. injection of ZoletilTM 50 (Virbac S.A) and xylazine hydrochloride. The skin over the coronal and sagittal sutures of anesthetized mice were sterilized with iodine followed by alcohol. An incision of 0.5 cm was made along the skin064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS over the midline to expose coronal and sagittal suture junctions. Animals were placed on the stereotaxic instrument (Stoelting, USA) for intracranial injection.2 × 105luciferase-expressing MDA-MB-436-luc tumor cells suspended in 2 μL L-15 media were injected into the right forebrain by positioning the needle at 2.0 mm lateral to the sagittal suture, 0.5-1.0 mm anterior to coronal suture with the injection depth precisely controlled at 3.0 mm. The injection was slowly proceeded over a one-minute period. Upon completing injection, the needle was retained for another minute. After the needle removal, the hole was sealed with bone wax and the incision was closed.

[0166] Animals received 5 mg / kg of Meloxicam by s.c. injection for analgesic therapy. Levofloxacin was added into the drinking water (0.1 mg / mL) for 3-7 days post the surgery.

[0167] The tumor growth was monitored by image analysis. The mice were randomized and treatment was started when the mean bioluminescent imaging (BLI) value reached approximately 20~30 photons / s^106for the efficacy study. Imaging Analysis of Tumor Development

[0168] Mice were injected intraperitoneally with 15 mg / mL (at 5 μl / g BW) of D-luciferin and anesthetized with 1-2% isoflurane inhalation once every week. At 10 minutes after luciferin injection, the mice were imaged using IVIS Lumina III (Perkin Elmer). Living Image software (Perkin Elmer) was used to compute regions of interest (ROI) and integrate the total bioluminescence signal in each ROI from the whole body. Bioluminescent imaging signals (photons / s) from ROI were quantified and used as an indicator of tumor growth and antitumor activity.

[0169] Bioluminescent imaging (BLI) signal decrease, expressed in %, was calculated using the following formula: BLI decrease (%) = (1-BLI treatment / BLI control) × 100%, where BLI treatment and BLIcontrolwere the mean BLI in the treated and the control groups, respectively, on a given day after tumor cell inoculation. Study-1

[0170] The anti-tumor efficacy of Compound 6 in MDA-MB-436-luc human breast cancer intracranial xenograft tumor model was evaluated. The grouping and dosing schedule are shown in Table 12.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Table 12 Group Na CompoundDosage Dosing volume Route of Dosing therapy(mg / kg) (µL / g)cadministrationdregimene1 8 Vehicle NA 10 p.o QD^39D 2 8 Compound 6 1 10 p.o QD^42D 3 8 Compound 6 3 10 p.o QD^42D 4 8 Compound 6 10 10 p.o QD^42D Note: a. N is the number of animals, 8 tumor-bearing nude mice per group; b. The vehicle is 0.5%MC in water; c. Dosing volume: adjust dosing volume based on body weight 10µL / g. In the event of 1) animals had lost significant body mass (emaciated); obvious body weight loss > 20% and did not recover after a dosing holiday; 2) animals could not take adequate food or water, euthanasia was performed on the affected animals. d. p.o. is oral administration; e. QD is once a day; continuous administration for 39 and 42 days for vehicle and Compound 6 treatment groups, respectively. NA means not applicable. 1) Results: 2) Mortality, Morbidity, and Body Weight Gain or Loss

[0171] In the vehicle group, obvious bodyweight loss and death were found from day 34 post tumor cell inoculation, which were most likely due to the intracranial tumor burden in mice.

[0172] Throughout 42-day treatment with Compound 6, no obvious body weight loss was observed, and no death or any signs of morbidity were found. 3) Tumor growth inhibition

[0173] Tumor growth inhibition information including BLI and BLI decrease (%) value compared to control group based on data of day 42 is shown in Table 13. Table 13 Group Compound Therapy Mean BLI (photons / s^106)a BbLI Decrease (%)1 Vehicle 807 ± 180 NA 2 Compound 6, 1 mg / kg 8 ± 3 99.0 3 Compound 6, 3 mg / kg 6 ± 2 99.3 4 Compound 6, 10 mg / kg 3 ± 0 99.6 Note: a. Mean ± SEM. b. vs control (that is vehicle). NA means not applicable.

[0174] In summary, all treatment groups showed significant tumor growth inhibition compared with the vehicle group. Compound 6 at the tested dose levels were well tolerated by tumor bearing mice during the study.064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Study-2

[0175] The anti-tumor efficacy of Compound 5 and 6 in MDA-MB-436-luc human breast cancer intracranial xenograft tumor model was evaluated. The grouping and dosing schedule are shown in Table 14. Table 14 Group Na CompoundDosage Dosing volume Route of Dosing therapy(mg / kg) (µL / g)cadministrationdregimene1 8 Vehicle NA 10 p.o QD^28D 2 8 Compound 5 1 10 p.o QD^28D 3 8 Compound 6 0.2 10 p.o QD^28D 4 8 Compound 6 0.5 10 p.o QD^28D 5 8 Compound 6 1 10 p.o QD^28D Note: a. N is the number of animals, 8 tumor-bearing nude mice per group; b. The vehicle is 0.5%MC in water; c. Dosing volume: adjust dosing volume based on body weight 10µL / g. In the event of 1) animals had lost significant body mass (emaciated); obvious body weight loss > 20% and did not recover after a dosing holiday; 2) animals could not take adequate food or water, euthanasia was performed on the affected animals. d. p.o. is oral administration; e. QD is once a day; continuous administration for 28 days. NA means not applicable. Results: 1) Mortality, Morbidity, and Body Weight Gain or Loss

[0176] In the vehicle group, obvious bodyweight loss (>20%) of one mouse, and then two on day 24 and on day 28 were found respectively. The three mice were euthanized.

[0177] In Group 3 (Compound 6 treatment group at 0.2 mg / kg), a mouse was found dead. Autopsy showed that there was serious food accumulation in the intestine and no tumor in the brain.

[0178] In Groups 2, 4, and 5, no obvious body weight loss was observed, and no death or any signs of morbidity were found. 2) Tumor growth inhibition

[0179] Tumor growth inhibition information including BLI and BLI decrease (%) value compared to control group based on data of day 28 is shown in Table 15. Table 15064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS Group Compound Therapy Mean BLI (photons / s^106)a BbLI Decrease (%)1 Vehicle 642±140 NA 2 Compound 5, 1 mg / kg 10±1 101.9% 3 Compound 6, 0.2 mg / kg 11±2 102.7% 4 Compound 6, 0.5 mg / kg 12±3 102.6% 5 Compound 6, 1 mg / kg 15±4 102.5% Note: a. Mean ± SEM. b. vs control (that is vehicle). NA means not applicable.

[0180] In summary, all treatment groups showed very significant tumor growth inhibition compared with the vehicle group. Example 20 Brain Penetration of Radiolabeled Compound 6 Following Oral Administration in Sprague Dawley Rats

[0181] The distribution of radiolabeled Compound 6 in blood, plasma and tissues was measured in Sprague Dawley rats following a single oral dose of 5 mg / 100 µCi / kg of [14C]Compound 6 to male and female intact rats at 1, 6, 12, and 24 h post-dose. The total radioactivity in plasma and whole brain is shown in the following tables. Table 16 Male Total Radioactivity (ng Eq. / g) Parameters Tissue 001h 006h 012h 024h TmaxCmaxAUC0-lastMean Mean Mean Mean(h) (ng Eq. / g)(h∙ng Eq. / g)Plasma 1587 1948 1139 206 6 1948 26962 Whole Brain 961 1066 650 120 6 1066 15316 Female Total Radioactivity (ng Eq. / g) Parameters Tissue 001h 006h 012h 024h TmaxCmaxAUC0-lastMean Mean Mean Mean (h) (ng Eq. / g) (h∙ng Eq. / g) Plasma 3340 2250 2210 530 1 3340 45465 Whole Brain 1519 1024 907 186 1 1519 19468

[0182] The results show that Compound 6 had good brain penetrability. Example 21064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS A pharmacokinetics study of Compound 6 in plasma and brain following single oral administration to rats

[0183] Compound 6 was formulated into a 0.5% methylcellulose / water uniform suspension, and administered to SD rats by gavage at 5 mg / kg. Each blood collection (about 0.2 mL per time point) was performed from jugular vein or other suitable site of each rat into pre-chilled EDTA-K2 tubes. The rats were euthanized at 1, 4 and 8 hours post-dose, respectively, using CO2 inhalation and then perfused with saline, and brain was collected from each rat. After collection, the brain was washed with cold saline, wiped dry and weighed. Brain was homogenized using homogenizing buffer (15 mM PBS (pH7.4): MeOH=2:1) at the ratio of 1:4 (1 g tissue with 4 mL buffer, the dilution ratio is 5). The tissue homogenate was kept at -60 °C or lower until LC-MS / MS analysis. Concentrations of Compound 6 in plasma and brain were determined by the LC-MS / MS method.

[0184] The concentrations, calculated AUC0-8h, and B / P ratio of Compound 6 are summarized in the following Table. Table 17 Time Concentration in Concentration in AUC0-8h in brain B / P point (h) plasma (ng / mL) brain (ng / g) (h·ng / g) * ratio 1 2497 338 0.14 4 3057 562 4003 0.18 8 2773 680 0.25 Note: * Compound concentration versus time data was analyzed by non-compartmental approaches using the WinNonlin software program, and AUC0-8h was calculated using the linear logarithmic trapezoidal method.

[0185] The result shows that Compound 6 has good exposure in brain in rats.

[0186] Having now fully described this disclosure, it will be understood by those of ordinary skill in the art that the same can be performed within a wide and equivalent range of conditions, formulations and other parameters without affecting the scope of the disclosure or any embodiment thereof. All patents, patent applications and publications cited herein are fully incorporated by reference herein in their entirety.

Claims

064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS WHAT IS CLAIMED IS:

1. A compound having the following structure of Formula (I):or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from the group of halo, alkyl and haloalkyl; R2is selected from the group of halo and alkyl; R3 is selected from the group of halo, alkyl and cyano; R4is selected from the group of alkyl, haloalkyl and cycloalkyl.

2. The compound of claim 1, wherein: (a) R1 is halo, C1-3 alkyl or halo C1-3 alkyl, or (b) R2 is halo or C1-3 alkyl, or (c) R3 is halo, C1-3 alkyl or cyano, or (d) R4 is C1-3 alkyl, halo C1-3 alkyl or C3-4 cycloalkyl, or any combination of (a) through (d).

3. The compound of claim 1, wherein: (e) R1 is F, methyl or trifluoromethyl, or (f) R2 is F, Cl or methyl, or (g) R3 is F, methyl or cyano, or (h) R4 is C1-3 alkyl, C1-3 haloalkyl or C3-4 cycloalkyl, or any combination of (e) through (h).

4. The compound of claim 1, wherein all of R1, R2, R3and R4of the compound are defined according to any one of rows 1-11 of the following table:064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS R1 R2 R3 R4 1. CH3F F CH32. CH3 F CN CH3 3. CH3 F F cyclopropyl 4. CF3F F CH35. F F CH3 CH3 6. F F F CH3 7. F F F CH2CH38. F F F CH2CHF2 9. F F F cyclopropyl 10. F Cl F CH311. F CH3 F CH3.

5. The compound of claim 2, wherein: (a) R1 is CH3, R2 is F, R3 is F, and R4 is CH3, or (b) R1 is CH3, R2 is F, R3 is F, and R4 is cyclopropyl, or (c) R1is F, R2is F, R3is CH3, and R4is CH3, or (d) R1is F, R2is F, R3is F, and R4is CH3, or (e) R1is F, R2is F, R3is F, and R4is CH2CHF2, or (f) R1is F, R2is Cl, R3is F, and R4is CH3.

6. A method for treating cancer in a subject comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-5, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, optionally wherein the cancer is a central nervous system (CNS) tumor.

7. The method of claim 6, wherein the subject has a central nervous system (CNS) tumor selected from meningioma, meningiosarcoma, ependymoma, astrocytoma, glioma,064423-520F05CN 2ndprovisional 238054 (IMP17-5 (EIK2003-P1)) -- 1PCWO PARP1 Inhibitors CNS glioblastoma, pineal cell tumor, aggressive pituitary tumors, pituitary carcinomas, germ cell tumor, sarcoma, glioblastoma multiforme, medulloblastoma, oligodendroglioma, craniopharyngioma, retinoblastoma, schwannoma, primary central nervous system lymphoma, brainstem glioma, pituitary adenoma, anaplastic astrocytoma, mixed glioma, primitive neuroectodermal tumor, hemangioblastoma, vestibular schwannoma, chordoma, spinal cord neurofibroma, lymphoma, optic nerve glioma, and dysembryoplastic neuroepithelial tumour.

8. A method for selectively inhibiting PARP1 activity relative to PARP2 activity in a subject comprising administering to the subject an effective amount of a compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof.

9. The method of claim 8, wherein the ratio of PARP1 activity inhibition to PARP2 activity inhibition in the subject is between 750 and 15,000.

10. A pharmaceutically acceptable composition comprising the compound of any one of claims 1-5, or a hydrate, or an isotope-substituted derivative, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable diluent, carrier or excipient.

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