PARP1 inhibitors for the treatment of diseases

Abstract

Disclosed herein are novel compounds, geometric isomers thereof, or pharmaceutically acceptable salts thereof, which are capable of inhibiting poly(ADP-ribose) polymerases. Also disclosed are methods of treating diseases or conditions associated with dysregulation or abnormal activity of PARP.

Description

PARP1 INHIBITORS FOR THE TREATMENT OF DISEASESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to Provisional Application No. 63 / 662,736, filed June 21, 2024 and Provisional Application No. 63 / 704,809, filed October 8, 2024, the disclosures of which is hereby incorporated by reference in the entirety.TECHNICAL FIELD

[0002] Disclosed herein are poly(ADP-ribose) polymerase inhibitors and their use in treating various diseases and conditions.BACKGROUND

[0003] The poly(ADP-ribose) polymerase (PARP) enzyme family consists of 17 members, with PARP1 being crucial for DNA repair. PARP1 attaches to single-strand breaks (SSBs) and, in an NAD+-dependent manner, catalyzes the creation of extensive branched chains of poly(ADP-ribose) (PAR) in a process called PARylation. This process recruits various DNA repair enzymes and allows PARP to detach from DNA. Current PARP inhibitors, which compete with NAD+, block PARP's enzymatic activity and prevent its release from damaged DNA, leading to a phenomenon known as PARP trapping. PARP trapping causes DNA double-strand breaks (DSBs) that require homologous recombination (HR) for accurate repair. Tumor cells with HR deficiencies due to BRCA1 or BRCA2 mutations are especially vulnerable to PARP inhibitors. Currently, there is still urgent need to discover potent PARP inhibitors for various clinical indications.SUMMARY

[0004] The compounds of this patent document address the need. Disclosed herein are novel PARP inhibitors that represent a new step toward the potential treatment of PARP deregulated diseases including, for example, pancreatic cancer, colorectal cancer, lung cancer, breast cancer and any other aberrant PARP associated diseases. In particular, the compounds can be used as inhibitors of PARPL The compounds can also be used with other therapeutic agents (such as immunotherapeutic, chemotherapeutic, and various targeted agents) for the treatment of diseases associated with PARP.

[0005] An aspect of the patent document provides a compound of formula I, a geometric isomer thereof, or a pharmaceutically acceptable salt thereof,whereinA is 5- or 6- membered aromatic ring;B is a 4-8 membered carbocyclic ring or heterocyclic ring;C is 9-10 membered bicyclic ring;L is C(O), O, SO2, Cl-3alkylene or NRn;M is H, deuterium, halogen, C3-6cycloalkyl or Cl-6alkyl;Ra in each instance is independently selected from the group consisting of deuterium, OC1- 6alkyL SCl-6alkyl, CN, OH, SH, halogen, NO2, N(Rm)2, C(0)0Rm, C(0)N(Rm)2. C(O)C1- 6alkyl, haloCl-6alkyl, haloCl-6alkyleneO, Cl-6alkyL hydroxyCl-6alkyl, dihydroxyCl- lOalkyl, C3-6cycloalkyl, C(=NCl-6alkyl)Cl-6alkyl, 0C(0)N(Rm)2, C(O)SRm, OC1- 6alkyleneOCl-6alkyl, OCl-6alkyleneO-haloCl-6alkyl, SCl-6alkyleneOCl-6alkyl, SC1- 6alkyleneSCl-6alkyl, OCl-6alkyleneSCl-6alkyl, SCl-6alkyleneO-haloCl-6alkyl, SC1- 6alkyleneS-haloCl-6alkyL OCl-6alkyleneS-haloCl-6alkyl, Cl-6alkylene-CN, OC1- 6alkylene-CN, SCl-6alkylene-CN, OCl-6alkylene-N(Rm)2, C2-6alkynyl, C2-6alkenyl, SO2N(Rm)2, NRmSO2Cl-6alkyl, Cl-6alkylSO2 (sulfone), S(O)OH, Cl-6alkylS(O) (sulfoxide), nitroso, and Cl-6alkylOSO2;Rb in each instance is independently selected from the group consisting of deuterium, OC1- 6alkyl, SCl-6alkyl, CN, OH, SH, halogen, N(Rm)2, C(0)0Rm, C(0)N(Rm)2, C(O)Cl-6alkyl, haloCl-6alkyl, Cl-6alkyl, hydroxyCl-6alkyl, C2-6alkynyl, and C2-6alkenyl;Rc in each instance is independently selected from the group consisting of deuterium, oxo, OCl-6alkyl, SCl-6alkyl, CN. OH, SH, halogen, NO2, N(Rm)2, C(0)0Rm. C(0)N(Rm)2, C(O)Cl-6alkyL haloCl -6alkyl, haloCl-6alkyleneO, Cl-6alkyl, hydroxyCl-6alkyl, dihydroxyCl-lOalkyl, C3-6cycloalkyL C(=NCl-6alkyl)Cl-6alkyl, 0C(0)N(Rm)2, C(O)SRm, OCl-6alkyleneOCl-6alkyl, OCl-6alkyleneO-haloCl-6alkyl, SCl-6alkyleneOCl- 6alkyl, SCl-6alkyleneSCl-6alkyl. OCl-6alkyleneSCl-6alkyl, SCl-6alkyleneO-haloCl- 6alkyL SCl-6alkyleneS-haloCl-6alkyL OCl-6alkyleneS-haloCl-6alkyl, C l-6alkylene-CN, OCl-6alkylene-CN, SCl-6alkylene-CN, OCl-6alkylene-N(Rm)2, C2-6alkynyl, C2-6alkenyl, SO2N(Rm)2, NRmSO2Cl-6alkyl, Cl-6alkylSO2 (sulfone), S(O)OH, Cl-6alkylS(O) (sulfoxide), nitroso, and Cl-6alkylOSO2;Rm each is independently hydrogen or Cl-6alkyl or halo-Cl-6alkyl;Rn is hydrogen, Cl-6alkyl, halo-Cl -6alkyl, C(O)Cl-6alkyl; m is 0, 1, 2, 3, 4 or 5; n is 0, 1, 2, or 3; and p is 0, 1, 2, 3, 4 or 5.

[0006] Another aspect of the patent document provides a pharmaceutical composition comprising the compound of formula I, geometric isomer thereof, or pharmaceutically acceptable salt thereof disclosed herein, and a pharmaceutically acceptable carrier.

[0007] Another aspect provides a method of treating a disease associated with abnormal or deregulated PARP enzyme, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of formula I, geometric isomer thereof, or pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof disclosed herein, wherein the disease is selected from the group consisting of neurodegenerative diseases, metabolic disease, cardiovascular diseases, autoimmune disease, and cancer.

[0008] Another aspect provides a method of inhibiting PARP enzyme, comprising contacting a cell with an effective amount of the compound of formula (I) or the pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof disclosed herein.DETAILED DESCRIPTION

[0009] This patent document discloses poly(ADP-ribose) polymerase (PARP) inhibitors and use of thereof for treating various diseases. The PARP inhibitors inhibit the activity or decreases the function of one or more of the over 15 different enzymes in the PARP family, which engage in a variety of cellular functions, including cell cycle regulation, transcription, and repair of DNA damage. In some embodiments, a PARP inhibitor inhibits PARP-1 and / or P ARP-2.

[0010] While the following text may reference or exemplify specific embodiments of a compound, substituent, or use thereof, it is not intended to limit the scope of the compound, substituent or its use to such particular references or examples. Various modifications may be made by those skilled in the art, in view of scientific and practical considerations, such as replacement of a substituent or treatment of other diseases.

[0011] The articles "a" and "an" as used herein refer to "one or more" or "at least one," unless otherwise indicated. That is, reference to any element or component of an embodimentby the indefinite article "a" or "an" does not exclude the possibility that more than one element or component is present.

[0012] The term "acyl” refers to -C(O)CH3. -C(O)CH2CH3, -C(O)CH2CH2CH3, - C(O)CH2CH2CH2CH3 and groups alike.

[0013] The term "alkyl" refers to a hydrocarbon or a hydrocarbon chain which may be either straight-chained or branched. The term "Ci-6 alkyl" refers to alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. Non-limiting examples include groups such as CH3, (CH2)2CH3, CH2CH(CH3)CH3, and the like. Similarly, the term "C2-5 alkyl" refers to alkyd groups having 2, 3, 4 or 5 carbon atoms. In some embodiments, an alkyl is used interchangely with alkylene and one of ordinary skill in the art could readily reconganize the meaning of the term in the context of a chemical structure.

[0014] The term '‘alkylene” refers to a divalent hydrocarbon or a hydrocarbon chain which may be either straight-chained or branched. Non-limiting examples include groups such as CH2, (CH2)2CH2, CH2CH(CH3)CH2, and the like. A Ci-3alky lene includes alkylenes with 1, 2 or 3 carbons such as CH2, (CH2)2, CHCH3,(CH2)3, and CH(CH3)CH2.

[0015] The term "cycloalky l" refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 ring carbons, for example 3 to 8 carbons, and as a further example 3 to 6 carbons, wherein the cycloalkyl group additionally is optionally substituted. Examples of cycloalkyl groups include, without limitation, cyclopropyl, cyclobuty l, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohepty l, and cycloocty l.

[0016] The term "ary l" group refers to a C6-14 aromatic moiety' comprising one to three aromatic rings, which is optionally substituted. Examples of ary l groups include, without limitation, phenyl, naphthyl, anthracenyl, fluorenyl, and dihydrobenzofuranyl. The term “aromatic ring” can also encompasses heteroaryl.

[0017] The term “alkeny” refers to a carbon chain containing a carbon-carbon double bond moiety. Non-limiting examples of alkenyl groups include ethylenyl, 1 -propenyl, allyl and 2-butenyl.

[0018] The term "alkynyl" group refers to a caron chain containing a carbon-carbon triple bond moiety7. Non-limiting examples of alkynyl groups include ethynyl, 1-propanyl, propargyl and 2-butynyl.

[0019] The term “haloalkyl” refers to a C6-ioalkyl chain, straight or branched, in which one or more hydrogen has been replaced by a halogen. Non-limiting examples of haloalkyls include CHF2, CFH2, CF3, CH2CHF2, CH2CH2CL CH2CF3, and CH2CH2F. In some embodiments, the alkyd in haloalkyl has 1 , 2, 3 or 4 carbons.

[0020] The term ‘'heteroalkyl” refers to a C6-ioalkyl group, straight or branched, wherein one or more carbon atoms in the chain are replaced by one or more heteroatoms selected from the group consisting of O, S, N and NRm. In some embodiments, the alkyl in heteroalkyl has 1 to 10 carbons. In some embodiments, the alkyl in heteroalkyd has 2. 3, 4 or more than 2 carbons.

[0021] The term '"geometric isomer’7refers to an isomer which has the same molecular formula and sequence of bonded atoms (constitution) of a reference compound, but differ in the spatial arrangement of atoms or groups around a rigid double bond structure. A geometric isomer is also known as a cis or trans isomer.

[0022] The term “hydroxyalkyl” refers to a C6-ioalkyl chain, straight or branched, wherein a carbon is substituted with a hydroxyl group. The carbon the hydroxyl is attached to is a primary carbon or secondary carbon. In some embodiments, the alkyl in hydroxylalkyl has 2, 3, 4 or more than 2 carbons.

[0023] The term “dihydroxyalkyl” refers to a C2-ioalkyl chain, straight or branched, wherein two carbons are each substituted with a hydroxyl group. In some embodiments, the alkyl in dihydroxylalkyl has 2, 3, 4 or more than 2 carbons.

[0024] The term “heterocyclyl” or “heterocyclic” group is a ring structure having from about 3 to about 12 atoms, for example 4 to 8 atoms, wherein one or more atoms are selected from the group consisting of N, O, and S. the remainder of the ring atoms being carbon. The heterocyclyl may be a monocyclic, a bicyclic, a spirocyclic or a bridged ring system. Examples of heterocyclic groups include, without limitation, epoxy, azetidinyl, aziridinyl, azocanyl, azepanyl, diazepanyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, oxazepanyl, pyrrolidinyl, pyrrolidinonyl. piperidinyl, piperazinyl. imidazolidinyl, thiazolidinyl, thiooxazepanyl, dithianyl, trithianyl, dioxolanyl. oxazolidinyL oxazohdinonyL decahydroquinolinyl, piperidonyl, 4-piperidinonyl, thiomorpholinyl, thiomorpholinyl 1,1 dioxide, morpholinyl, oxazepanyl, azabicyclohexanes, azabicycloheptanes and oxa azabiocycloheptanes. Specifically excluded from the scope of this term are compounds having adjacent annular O and / or S atoms.

[0025] The term “heteroaryl” refers to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6. 10. or 14 n electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to three heteroatoms per ring selected from the group consisting of N, O, and S. Examples of heteroaryl groups include acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl. benztetrazolyl. benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl. cinnolinyl. furanyl, furazanyl, imidazolinyl, imidazolyl, IH-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl. isoxazolyl, methylenedioxyphenyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl. 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole. pyridinyl, pyridyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyL pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-l,2,5-thiadiazinyl,1.2.3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl,1.2.3-triazolyl. 1,2.4-triazolyL 1.2.5-triazolyl, 1.3.4-triazolyl, and xanthenyl.

[0026] The term “halogen” refers to F, Cl. Br or I.

[0027] The term “subject” refers to humans or animals including for example sheep, horses, cattle, pigs. dogs, cats, rats, mice, birds, and reptiles. Preferably, the subject is a human or other mammal.

[0028] The term “effective amount” or “therapeutically effective amount” of a compound is an amount that is sufficient to ameliorate, or in some manner reduce a symptom or stop or reverse progression of a condition, or negatively modulate or inhibit activity. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective.

[0029] The term “hydrogen bond donor” refers to a group containing a hydrogen, which can be form a hydrogen bond with another electronegative atom such as F, N or O. Non-limiting examples of hydrogen bond donor include OH and NH2, which can share its hydrogen with electron rich atoms to form a hydrogen bond.

[0030] The term “hydrogen bond acceptor” refers to a group or atom rich in electrons, which can form a hydrogen bond with a hydrogen bond donor. Non-limiting examples of hydrogen bond acceptor include O, N and F.

[0031] The term “oxo” refers to an oxygen which is bonded to a carbon via a doule bond. For example, a carbon atom substituted with an oxo is a carbonyl group (C=O).

[0032] The term “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and / or bodily fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / nsk ratio.

[0033] The term “pharmaceutically acceptable carrier” refers to a chemical compound that facilitates the delivery or incorporation of a compound or therapeutic agent into cells or tissues.

[0034] The term “pharmaceutically acceptable salts” means salts of compounds of the present invention which are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. Non-limiting examples of such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; or with organic acids such as 1 ,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4'-methylenebis(3-hydroxy- 2-ene- 1 -carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene- 1 -carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxy naphthoic acid, lactic acid, laurylsulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid. o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acids, propionic acid, / Moluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, tertiarybutylacetic acid, and trimethylacetic acid. Pharmaceutically acceptable salts also include base addition salts which may be formed when acidic protons present arecapable of reacting with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide and calcium hydroxide. Non-limiting examples of acceptable organic bases include ethanolamine, diethanolamine, ethylenediamine, triethanolamine, tromethamine, and / V-methylglucamine. It should be recognized that the particular anion or cation forming a part of any salt of this invention is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag Helvetica Chimica Acta, 2002).

[0035] The term “pharmaceutical composition” refers to a mixture of a compound disclosed herein with other chemical components, such as diluents or additional carriers. The pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a pharmaceutical composition exist in the art including, but not limited to, oral, injection, aerosol, parenteral, intranasal, sublingual, inhalational, and topical administration. In some embodiments, pharmaceutically acceptable salts of the compounds disclosed herein are provided.

[0036] The term "treating" or "treatment" of any disease or condition refers, in some embodiments, to ameliorating the disease or disorder (i.e., arresting or reducing the development of the disease or at least one of the clinical signs and symptoms thereof). In some embodiments "treating" or "treatment" refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In some embodiments, "treating" or "treatment" refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. In some embodiments, "treating" or "treatment" refers to delaying the onset of the disease or disorder, or even preventing the same. “Prophylactic treatment” is to be construed as any mode of treatment that is used to prevent progression of the disease or is used for precautionary- purpose for persons at risk of developing the condition.

[0037] The PARP family of proteins consists of over 15 different enzymes, which engage in a variety of cellular functions, including cell cycle regulation, transcription, and repair of DNA damage. PARP enzymes can cleave NAD+, releasing nicotinamide, and successively add ADP-ribose units to form ADP-ribose polymers. Accordingly, activation of PARP enzy mes can lead to depletion of cellular NAD+ levels (e.g., PARPs as NAD+ consumers) and mediates cellular signaling through ADP-ribosylation of downstream targets.The role of PARP enzymes in DNA damage response (e.g. repair of DNA in response to genotoxic stress) has led to the compelling suggestion that PARP inhibitors may be useful anticancer agents.

[0038] PARP-1 is a zinc-finger DNA-binding enzyme that is activated by binding to DNA double or single strand breaks and is critical to the repair of single-strand DNA breaks through the base excision repair (BER) pathway. If such breaks persist unrepaired until DNA is replicated (which must precede cell division), then the replication itself can cause double strand breaks to form. Effective inhibition of PARP-1 leads to the accumulation of singlestrand breaks, which ultimately results in double-strand breaks. Usually such double-strand breaks are repaired by homologous recombination (HR), but in cells with defective HR, PARP inhibition can result in chromosomal instability, cell cycle arrest, and subsequent apoptosis. DNA is damaged thousands of times during each cell cycle, and that damage must be repaired. When subjected to enough damage at one time, the altered gene can cause the death of the cells. Normal cells that don't replicate their DNA as often as cancer cells, and that lack any mutated BRCA1 or BRCA2 still have homologous repair operating, which allows them to survive the inhibition of PARP. PARP inhibitors function by blocking PARP enzyme activity, which prevents the repair of DNA damage and ultimately may cause cell death. They also are believed to function by localizing PARP proteins at sites of DNA damage, which has relevance to their anti-tumor activity. The trapped PARP protein-DNA complexes are highly toxic to cells because they block DNA replication.

[0039] PARP-2 contains a catalytic domain and is capable of catalyzing a poly(ADP- ribosyl)ation reaction. PARP-2 displays auto-modification properties similar to PARP-1. The protein is localized in the nucleus in vivo and may account for the residual poly(ADP-ribose) synthesis observed in PARP-1 -deficient cells, treated with alkylating agents or hydrogen peroxide.

[0040] Studies have been directed to investigating the activity of PARP inhibitors, alone or in combination with other agents, as cancer therapeutics. PARP inhibitors may be particularly effective in treating cancers resulting from germ line or sporadic deficiency in the homologous recombination DNA repair pathway, such as BRCA-1, BRCA-2, and / or ATM deficient cancers. Additionally, simultaneous administration of genotoxic chemotherapy with PARP inhibition may enhance the killing effect of such chemotherapy by suppressing BER.

[0041] In some embodiments, the compounds disclosed herein exhibit improved selectivity for PARP1, which contributes to improved efficacy and reduced toxicity compared to other clinical PARP1 / 2 inhibitors. It is believed also that selective strong inhibition of PARP1 would lead to trapping of PARP1 on DNA, resulting in DNA double-strand breaks (DSBs) through collapse of replication forks in S-phase. Growing evidence suggests that a highly selective PARP1 inhibitor, which spares PARP2, could be an effective and well- tolerated treatment for cancer patients, either alone or in combination with immunotherapeutic, chemotherapeutic, and various targeted agents. For example, it may be combined with topoisomerase inhibitors such as Exatecan or their related ADCs. It may also be combined with Temozolomide (TMZ).

[0042] Compounds

[0043] An aspect of the disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof, which are capable of inhibiting the activity or decreases the function of one or more enzymes in the PARP family.Wherein:A is 5- or 6- membered aromatic ring;B is a 4-8 membered carbocyclic ring or heterocyclic ring;C is 9-10 membered bicyclic ring;L is C(O), O, SO2, Ci-4alkylene or NRn;M is H. deuterium, halogen, C3-6cycloalkyl or C1-6alkyl;Rain each instance is independently selected from the group consisting of deuterium, OC1-6alkyl, SCi-ealkyl, CN, OH, SH, halogen, NO2, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC 1-6alkyl, haloC1-6alkyleneO, C1-6alkyl, hydroxy C1-6alkyl, dihydroxyCi-ioalkyl, C3-6cycloalkyl, C(=NC1-6alkyl)C1-6alkyl, OC(O)N(Rm)2, C(O)SRm, OCi-6alkyleneOCi-6alkyl, OCi-ealkyleneO-hal oCi-ealkyl, SCi-ealkyleneOC1- 6alkyl, SC1-6alkyleneSCi ealkyl, OCi-ealkyleneSC1-6alkyl, SCi ealkyleneO-haloC1- 6alkyl, SC1-6alkyleneS-haloCi-ealkyl, OC1-6alkyleneS-haloC1-6alkyl, C1-6alkylene-CN,OC 1-6alkylene-CN, SC1-6alkylene-CN, OC1-6alkylene-N(Rm)2, C2-6alkynyl, C2-6alkenyl, SO2N(Rm)2, NRmSO2C1-6alkyl, C1-6alkylSCh (sulfone), S(O)OH, C1-6alkylS(O) (sulfoxide), nitroso, and C1-6alkylOSCh;Rbin each instance is independently selected from the group consisting of deuterium, OC1-6alkyl, SC1-6alkyl, CN, OH, SH, halogen, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, C1-6alkyl, hydroxyC1-6alkyl, C2-6alkynyl, and C2- 6alkenyl;Rcin each instance is independently selected from the group consisting of deuterium, oxo, OC1-6alkyl, SCi-ealkyl, CN, OH, SH, halogen, NO2, N(Rm)2, C(O)ORm, C(0)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO. C1-6alkyl, hydroxyC1- 6alkyl, dihydroxyC1-10alkyl, C3-6cycloalkyl. C(=NC1-6alkyl)C1-6alkyl, 0C(0)N(Rm)2, C(O)SRm, OC1-6alkyleneOC1-6alkyl, OC1-6alkyleneO-haloC1-6alkyl, SC1-6alkyleneOC1- 6alkyl, SC1-6alkyleneSCi ealkyl, OC1-6alkyleneSC1-6alkyl, SC1-6alkyleneO-haloC1- 6alkyl, SC1-6alkyleneS-haloC1-6alkyl, OC1-6alkyleneS-haloC1-6alkyl, C1-6alkylene-CN, OC1-6alkylene-CN, SC1-6alkylene-CN, OC1-6alkylene-N(Rm)2, C2-6alkynyl, C2-6alkenyl, SO2N(Rm)2, NRmSO2C1-6alkyl, C1-6alkylSO2(sulfone), S(O)OH, C1-6alkylS(O) (sulfoxide), nitroso, and C1-6alkylOSO2;Rmeach is independently hydrogen or C1-6alkyl or halo-C1-6alkyl;Rnis hydrogen, Ci-ealkyl, halo-C1-6alkyl, C(O)C1-6alkyl; m is 0, 1, 2, 3, or 4; n is 0, 1, 2, or 3; and p is 0, 1, 2, 3, 4 or 5.

[0044] In some embodiments, A is phenyl or 6-membered heteroaryl. In some embodiments, A pyridinyl or pyrimidinyl, which is optionally substituted with one, two, three or four Ras. In some embodiments, A optionally substituted pyridinyl, wherein the ring nitrogen is ortho, meta, or para to the M-substituted double bond moiety. In some embodiments, A optionally substituted pyridazine.

[0045] In some embodiments, A is pyridinyl optionally substituted with one or more Ras selected from OC1-6alkyl. SC1-6alkyl, CN, OH, SH. halogen. NO2, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, C1-6alkyl, hydroxyC1-6alkyl, and dihydroxyC 1-10alkyl. In some embodiments, the one or more Ras are selected from C(O)ORm, C(O)N(Rm)2, and C(O)Ci-ealkyl. In some embodiments, Rais C(O)N(Rm)2, whereinRmis C1-6alkyl or H. In some embodiments, Rais C(O)NHC i-4alkyl. In some embodiments, m is 1 and Rais C(O)N(Rm)2, wherein Rmis C1-6alkyl or H. In some embodiments, Rais C(O)NHCi-4alkyl is para to the M-substituted double bond moiety.

[0046] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof, wherein the compound is represented by Formula T-aWhereinRain each instance is independently selected from the group consisting of OC1-6alkyl, CN, halogen, N(Rm)2. C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl. haloC1-6alkyleneO. C1- 6alkyl, and hydroxyC1-6alkyl; m is 1, 2 or 3.

[0047] In some embodiments of the compound of formula I-a, at least one of (Ra)m is C(O)NHCi-6alkyl. In some embodiments, m is 1 and Rais C(O)N(Rm)2, wherein Rmis C1- 6alkyl or H. In some embodiments, Rais C(O)NHCi-4alkyl is para to the M-substituted double bond moiety.

[0048] In some embodiments of the compound of formula I, B is a 4-6 membered heterocyclic ring. The heteroatom can be at any chemically feasible position. In some embodiments, L is linked to a heteroatom of the B ring. In some embodiments, L is linked to a carbon atom of the B ring.

[0049] In some embodiments, the compound is represented by Formula I-b, wherein x is 1 or 2; and y is 1 or 2.

[0050] In some embodiments of the compound of formula I-b, n is 0 or 1. In some embodiments of the compound of formula I-b, x is 1, y is 1 or 2. In some embodiments of the compound of formula I-b, x is 2, y is 1 or 2.

[0051] In some embodiments, ring C comprises at least 1 ring nitrogen. In some embodiments, ring C comprises at least 1 ring nitrogen in each of the rings of the fused bicyclic system. In some embodiments, ring C is a 6-memered ring fused to another 6-memered ring. In some embodiments, one or both of the rings of the bicyclic system of C is aromatic. In some embodiments, at least one ring carbon of C is substituted with ox. In some embodiments of ring C, at least 1 ring nitrogen is directly bonded to ring carbon of C substituted with oxo.

[0052] In some embodiments. Rcin each instance is independently selected from the group consisting of oxo, OCi-ealkyl, SCi-ealkyl, CN, halogen, OH, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, and C1-6alkyl. In some embodiments, P is 1, 2 or 3. In some embodiments, one ring carbon of C is substituted with ox and C further comprises one or two substituents selected from OC1-6alkyl, CN. halogen, OH, N(Rm)2, haloC1-6alkyl, haloC1-6alkyleneO, and Ci-ealkyl. In some embodiments of ring C, at least 1 ring nitrogen is directly bonded to ring carbon of C substituted with oxo, and C further comprises a halogen and a C1-6alkyl.

[0053] In some embodiments, the compound is represented by Formula I-c,Wherein Raand Rband p are as defined above; Rcin each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, and C1-6alkyl; and p is 1, 2 or 3. Unless otherwise specified, (RC)Pof C can be positioned at one or both rings of the bicyclic system.

[0054] In some embodiments, the compound is represented by Formula I-d,Wherein Raand Rbare as defined above; Rcin each instance is independently selected from the group consisting of OCi-6alkyl, halogen, C(O)ORm, C(0)N(Rm)2, C(O)Ci-6alkyl, haloC1- 6alkyl, haloC1-6alkyleneO, and C1-6alkyl; and Wherein Rcin each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(0)N(Rm)2, C(O)C1- 6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, and C1-6alkyl; and p is 1, 2 or 3.

[0055] In some embodiments, the compound is represented by Formula I-e,whereinP is nitrogen or optionally substituted ring carbon (see Rcof Formula I for optional substituents);Q is nitrogen or optionally substituted ring carbon (see Rcof Formula I for optional substituents);U is N or optionally substituted carbon (see Rafor optional substituents);Rain each instance is independently selected from the group consisting of OC1-6alkyl, CN, halogen, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, C1- 6alkyl, and hydroxy C1-6alkyl;Rbis as defined above;Rcin each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloCi-ealkyl, haloC1-6alkyleneO, and C1-6alkyl;Subscript p is 0, 1, or 2.

[0056] In some embodiments of formula I-e, Rcin each instance is independently selected from the group consisting of halogen, haloC1-6alkyl, and C1-6alkyl. In some embodiments, n is 0 or 1.

[0057] In some embodiments, the compound is represented by Formula I-f,WhereinP is nitrogen or optionally substituted ring carbon (see Rcof Formula I for optional substituents);Q is nitrogen or optionally substituted ring carbon (see Rcof Formula T for optional substituents);T and U are each independently N or optionally substituted carbon (see Rafor optional substituents);Rclin each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, and C1- 6alkyl;Rc2in each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, and C1- 6alkyl;Rain each instance is independently selected from the group consisting of OCi-6alkyl, CN, halogen, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl. haloC1-6alkyl, haloC1-6alkyleneO, C1-6alkyl, and hydroxyC1-6alkyl;Ra’ is C(O)ORm, C(O)N(Rm)2, or C(O)C1-6alkyl;If only T is N. then m is 1, 2, or 3; if U and T are both N, then m is 0, 1 or 2.

[0058] In some embodiments of Formula I-f, n is 0 or 1. In some embodiments, T is N and U is optionally substituted ring carbon. In some embodiments, T and U are both N. In some embodiments, P is optionally substituted ring carbon and Q is nitrogen. In some embodiments, P is nitrogen and Q is or optionally substituted ring carbon.

[0059] In any embodiments disclosed herein where a stereocenter is present, the configuration can be R or S. The patent document also encompasses geometric isomers of the compounds disclosed herein. For instatnce. the alkene bonded to ring B can be cis or trans ingeometry. In some embodiments, one or more hydrogens of the compound of Formula I can be replaced with deuterium.

[0060] Further examples of formula I-f are shown below

[0061] In some embodiments, the compound is one of the following:

[0062] Pharmaceutical Composition and Kit

[0063] Another aspect of the patent specification provides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof disclosed herein and a pharmaceutically acceptable carrier, excipient, or diluent. Compounds described in this patent specification may be formulated by any method well known in the art and may be prepared for administration by any route, including, without limitation, parenteral, peroral, sublingual, buccal, intrathecal, transdermal, topical, subcutaneous, intramuscular, intraperitoneal, intranasal, intratracheal, or intrarectal.

[0064] Nonlimiting examples of pharmaceutically acceptable carriers include physiologically acceptable surface active agents, glidants. plasticizers, diluents, excipients, smoothing agents, suspension agents, complexing agents, film forming substances, and coating assistants. Preservatives, stabilizers, dyes, sweeteners, fragrances, flavoring agents, and the like may be provided in the pharmaceutical composition. For example, sodium benzoate, ascorbic acid and esters of p-hydroxybenzoic acid may be added as preservatives. In addition, antioxidants and suspending agents may be used. In various embodiments, alcohols, esters, sulfated aliphatic alcohols, and the like may be used as surface active agents. Suitable exemplary binders include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, and the like. Suitable exemplar)’ disintegrants include starch, carboxymethylcellulose, calciumcarboxymethylcellulose, croscarmellose sodium, sodium carboxymethylstarch, and the like. Suitable exemplary solvents or dispersion media include water, alcohol (for example, ethanol), polyols (for example, glycerol, propylene glycol, and polyethylene glycol, sesame oil, com oil, and the like), and suitable mixtures thereof that are physiologically compatible. Suitable exemplary7solubilizing agents include polyethylene glycol, propylene glycol, D-mannitol, benzylbenzoate, cyclodextrins, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate, and the like. Suitable exemplary suspending agents include surfactants such as stearyltriethanolamine, sodium laurylsulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glycerin monostearate, coconut oil, olive oil, sesame oil, peanut oil, soya and the like; and hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, and the like. Suitable exemplary isotonic agent includes sodium chloride, glycerin, D-mannose, and the like. Suitable exemplary buffer agents include buffer solutions of salts, such as phosphate, acetates, carbonates, and citrates. Suitable exemplary soothing agents include benzyl alcohol, and the like. Suitable exemplary antiseptic substances include para-oxybenzoic acid esters, benzethonium chloride, benzalkonium chloride, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, and the like. Suitable exemplary' antioxidants include sulfite salts, ascorbic acid, and the like. Suitable exemplary sealers include, but are not limited to HPMC (or hypromellose), HPC, PEG and combinations thereof. Suitable exemplary lubricants include magnesium stearate, calcium stearate, talc, colloidal silica, hardened oil and the like.

[0065] In further exemplary embodiments for solid preparations, carriers or excipients include diluents, lubricants, binders, and disintegrants. In exemplary' embodiments for liquid preparations, carriers include solvents, solubilizing agents, suspending agents, isotonic agents, buffer agents, soothing agents, and the like. Acceptable additional carriers or diluents for therapeutic use and the general procedures for the preparation of pharmaceutical compositions are well known in the pharmaceutical art, and are described, for example, in Remington’s Pharmaceutical Sciences, 18th Ed., Mack Publishing Co.. Easton, PA (1990), which is incorporated herein by reference in its entirety.

[0066] The compound of Formula I may also be in a pharmaceutically acceptable salt form. Examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hy drobromic acid, sulfuric acid, phosphoricacid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid. The compounds can also be administered as pharmaceutically acceptable quaternary' salts known by those skilled in the art, which specifically include the quaternary ammonium salt, wherein the counterion includes, for example, chloride, bromide, iodide. -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate).

[0067] A related aspect provides a kit, which includes a compound of Formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof and an instruction for treating or preventing certain diseases or conditions. In some embodiments, the kit further includes an additional secondary therapeutic agent.

[0068] In some embodiments, the secondary agent is an anti-cancer agent. In any of the methods or kit described herein, the administration or inclusion of a secondary agent having a cytotoxic effect on a cancer cell is contemplated. A cytotoxic effect refers to the depletion, elimination and / or the killing of target cells (i.e., tumor cells). The cytotoxic agent may be at least one selected from the group consisting of an antimetabolite, a mitotic inhibitor, an alkylating agent (e.g. temozolomide (TMZ)). an antibody-based EGFR inhibitor, an antibody based HER2 / 3 inhibitor, an angiogenesis inhibitor, a mTOR inhibitor, a CDK4 and CDK6 inhibitor or an aromatase inhibitor. The combination may include at least two cytotoxic agents. For example, the combination may include at least 2, at least 3. or at least 4 selected from the group consisting of an antimetabolite, a mitotic inhibitor, an alkylating agent, an angiogenesis inhibitor, or all of them.

[0069] The antimetabolite may be a drug that inhibits DNA synthesis in cells by suppressing formation of purines or pyrimidines, which are bases of a nucleotide. In one embodiment, the antimetabolite may be selected from the group consisting of Capecitabine, 5- Fluorouracil, Gemcitabine. Pemetrexed, Methotrexate, 6-Mercaptopurine, Cladribine, Cytarabine, Doxifludine. Floxuridine, Fludarabine, Hydroxycarbamide, decarbazine, hydroxyurea, and asparaginase. In a more specific embodiment, the antimetabolite is a base analog, with the term base analogs herein including nucleotide and nucleoside analogs in addition to purine base analogs such as 5 -fluorouracil.

[0070] The mitotic inhibitor may be a microtubule-destabilizing agent, a microtubulestabilizing agent, or a combination thereof. The mitotic inhibitor may be selected from taxanes, vinca alkaloids, epothilone, or a combination thereof. In a specific embodiment, the mitotic inhibitor is a taxane, for example including but not limited to, paclitaxel, docetaxel and cabazitaxel. In another specific embodiment, the mitotic inhibitor is a vinca alkaloid or its derivative, for example including but not limited to, vinblastine, vincristine, vinflunine, vinorelbine, vincaminol. vinbumine. vineridine and vindesine.

[0071] The mitotic inhibitor may be selected from BT-062, HMN-214. eribulin mesylate, vindesine, EC-1069, EC-1456, EC-531, vintafolide, 2-methoxyestradiol, GTx-230, trastuzumab emtansine (T-DM1), crolibulin, D1302A-maytansinoid conjugates IMGN-529, lorvotuzumab mertansine, SAR-3419, SAR-566658, IMP-03138, topotecan / vincristine combinations, BPH-8, fosbretabulin tromethamine, estramustine phosphate sodium, vincristine, vinflunine. vinorelbine, RX-21101, cabazitaxel. STA-9584. vinblastine, epothilone A, patupilone, ixabepilone, Epothilone D, paclitaxel, docetaxel, DJ-927, discodermolide, eleutherobin, and pharmaceutically acceptable salts thereof or combinations thereof.

[0072] Non-limiting examples of checkpoint inhibitors include those that target PD-1, PD-L1, CTLA4 and TIGIT (T cell immunoglobulin and ITIM domain). Further examples include Ipilimumab (Yervoy®; blocking a checkpoint protein called CTLA-4); pembrolizumab (Keytruda®), Cemiplimab (Libtayo) and nivolumab (Opdivo®) (targeting another checkpoint protein called PD-1); atezolizumab (Tecentriq®), Avelumab (Bavencio), and Durvalumab (Imfinzi) (targeting PD-L1); MK-7684, Etigilimab / OMP-313 M32, Tiragolumab / MTIG7192A / RG-6058, BMS-986207, AB-154 and ASP-8374 (targeting TIGIT), and V-domain Ig suppressor of T cell activation (VISTA).

[0073] The EGFR inhibitors may be selected from erlotinib, gefitinib, lapatinib, canetinib, pelitinib, neratinib, (R,E)-N-(7-chloro-l-(l-(4-(dimethylamino)but-2-enoyl)azepan- 3-yl)-lH-benzo[d]imidazol-2-yl)-2 -methylisonicotinamide, Trastuzumab, Margetuximab, panitumumab, matuzumab, necitumumab, pertuzumab, nimotuzumab, zalutumumab, cetuximab, icotinib, afatinib. and pharmaceutically acceptable salt thereof. In one embodiment the EGFR inhibitor may be an antibody based EGFR inhibitor such as cetuximab and in another embodiment, it is necitumumab and yet in another embodiment it is pantitumumab. The molecularly targeted agent may be an anti-EGFR family antibody or a complex including the anti -EGFR family antibody. The anti-EGFR family antibody may be an anti-HERl antibody, an anti-HER2 antibody, or an anti-HER4 antibody.

[0074] Further examples of agents for chemotherapy include SHP2 inhibitors (e.g. RMC-4550 and RMC-4630), phosphatase inhibitors (e.g. Tautomycin). CDK 4 / 6 inhibitors (abemaciclib (Lilly), palbociclib (Pfizer)), protein-protein interaction disruptors (BI 1701963), HSP90 inhibitor, tubulin inhibitor, apoptosis inhibitor, chemopreventative agent, and therapies targeting PBK / AKT / mTOR pathway.

[0075] Antibody-drug conjugates have emerged as a breakthrough approach to the development of cancer therapeutics. Cancer is one of the leading causes of deaths in the world. Antibody-drug conjugates (ADCs) comprise monoclonal antibodies (MAbs) that are covalently linked to cell-killing drugs. This approach combines the high specificity of MAbs against their antigen targets with highly potent cytotoxic drugs, resulting in “armed’’ MAbs that deliver the pay load (drug) to tumor cells with enriched levels of the antigen. Targeted delivery of the drug also minimizes its exposure in normal tissues, resulting in decreased toxicity and improved therapeutic index. The approval of two ADC drugs, ADCETRIS® (brentuximab vedotin) in 2011 and KADCYLA® (trastuzumab emtansine or T-DM1) in 2013 by FDA validated the approach. Another example is Trastuzumab duocarmazine. There are currently more than 30 ADC drug candidates in various stages of clinical trials for cancer treatment. As antibody engineering and linker-payload optimization are becoming more and more mature, the discovery and development of new ADCs are increasingly dependent on the identification and validation of new targets that are suitable to this approach and the generation of targeting MAbs. Two criteria for ADC targets are upregulated / high levels of expression in tumor cells and robust internalization.

[0076] Method of Treating Diseases

[0077] Another aspect of the patent specification provides for methods for treating a disease or condition including for example cancer, neurodegenerative disorders and viral infections. The method includes administering to a subject in need thereof the compound of formula I, a geometrical isomer thereof, a pharmaceutically acceptable salt thereof, or a corresponding pharmaceutical composition disclosed herein.

[0078] In some embodiments, the disease is associated with abnormal or deregulated PARP1. In some embodiments, the compound disclosed herein is selective for PARP1 over PARP2 by at least 50 folds, at least 80 folds, at least 100 folds, at least 150 folds, or at least 200 folds.

[0079] In some embodiments, the disease treatable with the methods disclosed herein is cancer including for example adenocarcinoma, adenocarcinoma of the lung, acute myeloid leukemia (“AML”), adrenocortical carcinoma, anal cancer, appendiceal cancer, B-cell derived leukemia, B-cell derived lymphoma, bladder cancer, brain cancer (e.g. glioblastoma), breast cancer (e.g., triple negative breast cancer (TNBC)), cancer of the fallopian tube(s), cancer of the testes, cerebral cancer, cervical cancer, choriocarcinoma, chronic myelogenous leukemia, colon adenocarcinoma, colon cancer, colorectal cancer, diffuse large B-cell lymphoma (“DLBCL”), endometrial cancer, epithelial cancer, esophageal cancer, Ewing's sarcoma, follicular lymphoma (“FL”), gall bladder cancer, gastric cancer, gastrointestinal cancer, glioma, head and neck cancer, a hematological cancer, hepatocellular cancer, Hodgkin's lymphoma / primary mediastinal B-cell lymphoma, kidney cancer, kidney clear cell cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, Merkel cell carcinoma, mesothelioma, monocytic leukemia, multiple myeloma, myeloma, a neuroblastic- derived CNS tumor, non-small cell lung cancer (NSCLC), oral cancer, ovarian cancer, ovarian carcinoma, pancreatic cancer, peritoneal cancer, primary’ peritoneal cancer, prostate cancer, relapsed or refractory' classic Hodgkin's Lymphoma (cHL), renal cell carcinoma, rectal cancer, salivary gland cancer (e.g., a salivary gland tumor), sarcoma, skin cancer, small cell lung cancer, small intestine cancer, squamous cell carcinoma of the anogenital region, squamous cell carcinoma of the esophagus, squamous cell carcinoma of the head and neck (SCHNC), squamous cell carcinoma of the lung, stomach cancer, T-cell derived leukemia, T-cell derived lymphoma, thymic cancer, a thymoma, thyroid cancer, uveal melanoma, urothelial cell carcinoma, uterine cancer, uterine endometrial cancer, uterine sarcoma, vaginal cancer, or vulvar cancer.

[0080] In some embodiments, the disease treatable yvith the methods disclosed herein is selected from endometrial cancer, uterine sarcoma, breast cancer, ovarian cancer, cervical cancer, fallopian tube cancer, primary peritoneal cancer, colon cancer, gastrointestinal cancer, squamous cell carcinoma of the anogenital region, melanoma, renal cell carcinoma, lung cancer, non-small cell lung cancer, squamous cell carcinoma of the lung, stomach cancer, bladder cancer, gall bladder cancer, liver cancer, thyroid cancer, laryngeal cancer, salivary gland cancer, esophageal cancer, head and neck cancer, squamous cell carcinoma of the head and neck, prostate cancer, lung cancer, pancreatic cancer, mesothelioma, sarcoma, or hematological cancer.

[0081] In some embodiments, the disease treatable with the methods disclosed herein is selected from bladder cancer, breast cancer, cancer of the fallopian tube(s), cholagiocarcinoma, colon adenocarcinoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, gastric cancer, kidney clear cell cancer, lung cancer, mesothelioma, ovarian cancer, pancreatic cancer, peritoneal cancer, prostate cancer, uterine endometrial cancer, or uveal melanoma.

[0082] In some embodiments, the disease treatable with the methods disclosed herein is breast cancer or triple negative breast cancer (TNBC). In some embodiments, the cancer is lung cancer or non-small cell lung cancer (NSCLC). In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is gynecological cancer (e.g., ovarian cancer, cervical cancer, fallopian tube cancer, or primary' peritoneal cancer). In some embodiments, the cancer is a recurrent cancer.

[0083] In some embodiments, the subject is a cancer patient identified to have deficiency in at least one gene involved in the homologous recombination repair (HRR) pathway, wherein the at least one gene involved in the HRR pathway is not BRC Al or BRCA2. In some embodiments, the subject has a deficiency in at least one gene selected from the group consisting of RFC2, XRCC6, POLD2, PCNA, RPA1, RPA2, ERCC3, UNG, ERCC5, MLH1, LIG1, MSH6, POLD4, RFC5, DDB2 / / / LHX3. POLDI, FANCG. POLB, XRCC1, MPG, ERCC1, TDG, FANCA, RFC4, RFC3, APEX2, RADI. EXO1, FEN1, MLH3. MGMT. RAD51, XRCC4, RECQL, ERCC8, FANCC, OGGI, MRE1 1A, RAD52, WRN, XPA, BLM, MSH3, POLE2, RAD51C, LIG4, ERCC6, LIG3, RAD17, XRCC2, MUTYH, RFC1, RAD50, DDB1, XRCC5, PARP1, POLE3, XPC. MSH2, RPA3. MBD4, NTHL1, PMS2 / / / PMS2CL, UNG2. APEX1. ERCC4, RECQL5, MSH5. POLD3, ERCC2, RECQL4, PMS1, ZFP276, POLE, XRCC3, NBN, SMUG1, FANCF, NEILL FANCE, ATM, ATR, BAP1, BARD1, BRIP1, PALB2, RAD51B, RAD51D, RAD54L, TP53, RBI, and combinations thereof.

[0084] In some embodiments, the subject is a cancer patient who has no germline or sporadic mutation in BRCA1 and no germline or sporadic mutation in BRCA2. In some embodiments, a patient (e.g., a cancer patient) has no germline mutation in BRCA1 and / or BRCA2. In some embodiments, a patient (e.g., a cancer patient) has no sporadic mutation in BRCA1 and / or BRCA2. In some embodiments, a patient (e.g., a cancer patient) has no tumor BRCA1 and / or BRCA2 mutations.

[0085] In some embodiments, the subject is a cancer patient who has at least one germline mutation in BRCA1 and / or BRCA2. In some embodiments, a patient (e.g., a cancer patient) has at least one sporadic mutation in BRCA1 and / or BRCA2. In some embodiments, a patient (e.g., a cancer patient) has at least one germline or sporadic mutation in BRCA1, and at least one germline or sporadic mutation in BRCA2. In some embodiments, a patient (e.g., a cancer patient) has at least one tumor BRCA1 and / or BRCA2 mutation.

[0086] In some embodiments, the subject has previously been treated with one or more different cancer treatment modalities. In some embodiments, a patient (e.g., a cancer patient) has previously been treated with one or more of radiotherapy, chemotherapy, or immunotherapy. In some embodiments, a patient (e.g., a cancer patient) has been treated with one, two, three, four, or five lines of prior therapy. In some embodiments, a patient (e.g., a cancer patient) has been treated with one or two lines of prior therapy. In some embodiments, a patient (e.g.. a cancer patient) has been treated with one line of prior therapy. In some embodiments, a patient (e.g., a cancer patient) has been treated with two lines of prior therapy. In some embodiments, a prior therapy is cytotoxic therapy. In some embodiments, a prior therapy is platinum-based chemotherapy.

[0087] In some embodiments, a patient (e.g., a cancer patient) has undergone at least one cycle of a platinum-based chemotherapy. In some embodiments, a patient (e.g.. a cancer patient) has undergone at least two cycles of a platinum-based chemotherapy. In some embodiments, a cancer is platinum-sensitive. In some embodiments, a patient (e g., a cancer patient) has a complete response or a partial response to the most recent cycle of platinumbased chemotherapy. In some embodiments, a patient (e g., a cancer patient) has a complete response of a partial response to the penultimate cycle of platinum-based chemotherapy. In some embodiments, administration of a PARP inhibitor is commenced within 8-weeks of the end of the last cycle of platinum-based chemotherapy. In some embodiments, a cancer is recurrent lung cancer (e.g., a recurrent non-small cell lung cancer (NSCLC)). In some embodiments, a cancer patient has undergone at least two cycles of a platinum-based chemotherapy. In some embodiments, a cancer is platinum-sensitive. In some embodiments, a cancer patient has a complete response to the platinum-based chemotherapy. In some embodiments, a cancer patient has a partial response to the platinum-based chemotherapy.

[0088] In some embodiments, a cancer is recurrent ovarian cancer, fallopian tube cancer, or primary peritoneal cancer. In some embodiments, a cancer patient has undergone at least one cycle of a platinum-based chemotherapy. In some embodiments, a cancer patient hasundergone at least two cycles of a platinum-based chemotherapy. In some embodiments, a cancer is platinum-sensitive. In some embodiments, a cancer patient has a complete response to the platinum-based chemotherapy. In some embodiments, a cancer patient has a partial response to the platinum-based chemotherapy. In some embodiments, administration of a PARP inhibitor is commenced within 8-weeks of the end of the last cycle of platinum-based chemotherapy.

[0089] In some embodiments, the methods disclosed herein comprise the administration of a second agent. Nonlimiting examples of the second agent are as described above. In some embodiments, a one or more additional therapeutic agent is a chemotherapeutic agent. In some embodiments, a chemotherapeutic agent is a platinum agent (e.g., cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, or the like). In some embodiments, a one or more additional therapeutic agent is an immune checkpoint inhibitor. In some embodiments, one, two. or three immune checkpoint inhibitors are administered. In some embodiments, an immune checkpoint inhibitor is an agent that inhibits programmed death- 1 protein (PD-1) signaling, T-cell immunoglobulin domain and mucin domain 3 (TIM-3), cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), lymphocyte activation gene-3 (LAG-3), or T-cell immunoglobulin and ITIM domain (TIGIT). In some embodiments, an immune checkpoint inhibitor is an antibody. In some embodiments, an immune checkpoint inhibitor is a T-cell immunoglobulin domain and mucin domain 3 (TIM- 3) inhibitor. In some embodiments, an immune checkpoint inhibitor is a cytotoxic T- lymphocyte-associated protein 4 (CTLA-4) inhibitor. In some embodiments, a PD-1 signaling inhibitor is an antibody (e.g., BGB-A317, BI 754091, IBI308, INCSHR-1210, JNJ-63723283, JS-001, MEDI-0680, MGA-012, mvolumab, PDR001, pembrolizumab, PF-06801591, REGN- 2810, TSR-042, atezolizumab, avelumab, CX-072, durvalumab, FAZ053, LY3300054, PD-L1 millamolecule, or derivatives thereof). In some embodiments, a PD-1 signaling inhibitor is an anti-PD-Ll / L2 agent. In some embodiments, an anti-PD-Ll / L2 agent is an antibody (e.g., atezolizumab, avelumab, CX-072, durvalumab, FAZ053, LY3300054, PD-L1 millamolecule, or derivatives thereof).

[0090] In some embodiments, the disease treatable with the methods disclosed herein is a neurodegenerative disease selected from for example Alzheimer’s disease, Lewy body dementia, frontotemporal dementia, traumatic brain injury’, prion diseases, Huntington’s disease, Parkinson’s disease, chronic traumatic encephalopathy, amyotrophic lateral sclerosis, mixed dementias, vascular dementia, hydrocephalus, and amyotrophic lateral sclerosis.

[0091] In some embodiments, the disease treatable with the methods disclosed herein is a metabolic disease or disorder (??). Examples include rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease and ulcerative colitis.

[0092] In some embodiments, the disease treatable with the methods disclosed herein is a inflammatory and autoimmune disease. Examples include diabetic complications, such as diabetic neuropathy and nephropathy, due to increased DNA damage and oxidative stress.

[0093] In some embodiments, the disease treatable with the methods disclosed herein is a cardiovascular disease including ischemia-reperfusion injury7and heart failure.

[0094] Also disclosed in this patent document is the use of a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof to treat a disease or condition. This patent document further provides a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof for use in the treatment of a disease or condition. The disease or condition, the means of administration, the dosage form and formulation, and the additional agents are the same as in the methods described herein.

[0095] Another aspect of the patent document discloses a method of inhibiting a PARP enzyme. The method includes contacting the enzyme or a cell containing it with an effective amount of a compound of Formula T. geometric isomer thereof or a pharmaceutically acceptable salt thereof. The PARP may be purified or crude, and may be present in a cell, tissue, or a subject. Thus, such a method encompasses inhibition of PARP activity both in vitro and in vivo. In some embodiments, the PARP is in a cell. In some embodiments, the PARP is expressed at normal levels in a subject, bin the subject would benefit from PARP inhibition. In some embodiments, the PARP is in a subject known or identified as having abnormal PARP activity (e.g., overexpression). Tn some embodiments, the method selectively inhibits PARP1 over PARP2. The selectivity can lead to safe and effective treatment of cancer such as brain tumors and neuroblastoma, both as monotherapies and m combination with one or more other agents.

[0096] It will be understood by one of ordinary skill in the art that inhibition of PARP does not necessarily require that all of the PARP be occupied by an inhibitor at once. Exemplary levels of inhibition of PARP include at least 10% inhibition, about 10% to about 25% inhibition, about 25% to about 50% inhibition, about 50% to about 75% inhibition, at least 50% inhibition, at least 75% inhibition, about 80% inhibition, about 90% inhibition, and greaterthan 90% inhibition. In some embodiments, the amount of the compound is selected so that one or more of the aforementioned levels of inhibition can be achieved. In some embodiments, the PARI’ is PARP1 and / or PARP2.

[0097] Administration Regimen

[0098] The compound of Formula I. or a pharmaceutically acceptable salt thereof or a pharmaceutically composition thereof for the methods or kit described herein described herein may be administered to the subject by any suitable means. Non-limiting examples of methods of administration include, among others, (a) administration though oral pathways, which administration includes administration in capsule, tablet, granule, spray, syrup, or other such forms; (b) administration through non-oral pathways such as rectal, vaginal, intraurethral, intraocular, intranasal, or intraauricular, which administration includes administration as an aqueous suspension, an oily preparation or the like or as a drip, spray, suppository, salve, ointment or the like; (c) administration via injection, subcutaneously, intraperitoneally, intravenously, intramuscularly, intradermally, intraorbitally, intracapsularly, intraspinally, intrastemally, or the like, including infusion pump delivery; as well as (d) administration topically; as deemed appropriate by those of skill in the art for bringing the active compound into contact with living tissue.

[0099] Advantageously, the compound of Formula I, or a pharmaceutically acceptable salt thereof or a pharmaceutically composition thereof for administrations described above are prepared into dosage forms in a unit dose suited to fit a dose of the active ingredients. Such dosage forms in a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc.

[0100] In exemplary embodiments of the pharmaceutical composition of the compound of Formula I, or a pharmaceutically acceptable salt thereof for oral administration, the composition can be a tablet, coated tablet, capsule, caplet, cachet, lozenges, gel capsule, hard gelatin capsule, soft gelatin capsule, troche, dragee, dispersion, powder, granule, pill, liquid, an aqueous or non-aqueous liquid suspension, an oil-in-liquid or oil-in-water emulsion, including sustained release formulations that are known in the art. For pediatric and geriatric applications, suspensions, syrups and chewable tablets are especially suitable.

[0101] The therapeutically effective amount (dosage) of the compound of Formula I, or a pharmaceutically acceptable salt thereof required will depend on the route of administration, the species (human or animal), and the physical characteristics of the particularsubject or patient being treated. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize. More specifically, a therapeutically effective amount means an amount of compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the patient or animal being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0102] In non-human animal studies, applications of potential products are commenced at higher dosage levels, with dosage being decreased until the desired effect is no longer achieved or adverse side effects disappear. The dosage may range broadly, depending upon the desired effects and the therapeutic indication. Typically, dosages may be about 10 pg / kg to about 100 mg / kg body weight, preferably about 100 pg / kg to about 10 mg / kg body weight. Alternatively, dosages may be based and calculated upon the surface area of the animal, as understood by those of skill in the art.

[0103] The exact formulation, route of administration and dosage for the pharmaceutical compositions can be chosen by the individual physician in view of the patient’s condition, (see e.g., Fingl et al. 1975, in “The Pharmacological Basis of Therapeutics’", which is hereby incorporated herein by reference in its entirety, with particular reference to Ch. 1, p. 1). In some embodiments, the dose range of the compound of Formula I or a pharmaceutically acceptable salt thereof administered to the subject or patient can be from about 0.5 to about 1000 mg / kg of their body weight. The dosage may be a single one or a series of two or more given in the course of one or more days, as is needed by the patient. In instances where human dosages for compounds have been established for at least some conditions, those same dosages, or dosages that are about 0.1% to about 500%, more preferably about 25% to about 250% of the established human dosage may be used.

[0104] It should be noted that the attending physician would know how to and when to terminate, interrupt, or adjust administration due to side-effects, toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response was not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary' with the severity' of the condition to be treated and to the route of administration. The severity of the condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency will also vary’ according to the age, body weight, and response of the individualpatient. A program comparable to that discussed above may also be used in veterinary medicine.

[0105] Although the exact dosage will be determined on a drug-by-drug basis, in most cases, some generalizations regarding the dosage can be made. The daily dosage regimen for an adult human patient may be, for example, a peroral dose of about 0.01 mg to 2000 mg of the active ingredient, preferably from about 0.01 mg to about 500 mg. In other embodiments, an intravenous, subcutaneous, or intramuscular dose of the active ingredient of about 0.01 mg to about 100 mg. preferably about 0.01 mg to about 60 mg is used. In cases of administration of a pharmaceutically acceptable salt, dosages may be calculated as the freebase. In some embodiments, the composition is administered 1 to 4 times per day. Alternatively, a compound of Formula I or a pharmaceutically acceptable salt thereof may be administered by continuous intravenous infusion, preferably at a dose of up to about 1000 mg per day. As will be understood by those of skill in the art. in certain situations it may be necessary to administer a compound of Formula I or a pharmaceutically acceptable salt thereof disclosed herein in amounts that exceed, or even far exceed, the above-stated, preferred dosage range in order to effectively and aggressively treat particularly intractable diseases or conditions. In some embodiments, a compound of Formula I or a pharmaceutically acceptable salt thereof will be administered for a period of continuous therapy, for example for a week or more, or for months or years.

[0106] In some embodiments, a compound of Formula I or a pharmaceutically acceptable salt thereof is formulated into a dosage form for release for a period of 1 to 12, ty pically 3 to 12 hours, more typically 6-12 hours after administration. In some embodiments, the oral pharmaceutical compositions described herein may be administered in single or divided doses, from one to four times a day. The oral dosage forms may be conveniently presented in unit dosage forms and prepared by any methods well known to those skilled in the art of pharmacy.

[0107] A compound of Formula I or a pharmaceutically acceptable salt thereof can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of the compound may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of such studies are often predictive of toxicity in animals, such as mammals, or more specifically, humans. Alternatively, the toxicity may be determined in an animal model (such as mice, rats, rabbits, or monkeys) using known methods. The efficacy of a particular compound may be established using severalrecognized methods, such as in vitro methods, animal models, or human clinical trials. Recognized in vitro models exist for nearly every class of condition. Similarly, acceptable animal models may be used to establish the efficacy of chemicals to treat such conditions. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the art to choose an appropriate model, dose, and route of administration, and dosing regime. Of course, human clinical trials can also be used to determine the efficacy of a compound of Formula I or a pharmaceutically acceptable salt thereof in humans.

[0108] A compound of Formula I or a pharmaceutically acceptable salt thereof may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions comprising a compound of Formula I or a pharmaceutically acceptable salt thereof formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0109] The following non-limiting examples serve to further illustrate the embodiments of the present disclosure.

[0110] Examples

[0111] Example 1 : compound synthesis

[0112] Compounds disclosed herein can be prepared via chemistry known in the organic synthesis. The scheme below illustrates a synthesis route to a compound under Formula I. Isomers can be separated via procedures including chromatography column, recrystallization and other methods known in the field of organic synthesis. The cross in the structures below represents a double bond which can be in a cis or trans configuration. The two isomers can be isolated with purification techniques well-known in the field of organic chemi stry.

[0113] Synthesis scheme for Int. B

[0114] Synthesis scheme for Int. D

[0115] A solution of 8-fluoro-7-(hydroxymethyl)-3-methyl-lH-quinoxalin-2-one (1 g, 4.803 mmol, 1 equiv) and POCI3 (20 mL) in DCM (20 mL) was stirred at 25°C for 16h. The precipitated solids were collected by filtration and washed with DCM (3x20 mL). This resulted in 7-(chloromethyl)-8-fluoro-3-methyl-lH-quinoxalin-2-one (900 mg) as a pink solid.LCMS:(ES,m / z):227 [M+l]+

[0116] A solution of 6-bromopyridazine-3 -carboxylic acid la (1 g, 4.926 mmol, 1 equiv) and CDI (0.80 g, 4.926 mmol, 1 equiv) in THF (20 mL) was stirred at 70°C for Ih. To the above mixture was added methylamine (2M in THF) (2.46 mL, 4.926 mmol, 1 equiv) at 70°C. The resulting mixture was stirred at 70°C for additional 2h. The mixture was allowed to cool down to 25 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 10% to 80% gradient in 10 min; detector, UV 254 nm to afford 6-bromo-N-methylpyridazine-3-carboxamide lb (200 mg, 18.79%yield) as a colorless oil. LCMS- :(ES,m / z):216[M+l]+

[0117] A solution of 6-bromo-N-methylpyridazine-3-carboxamide lb (500 mg, 2.314 mmol, 1 equiv), tert-butyl 3-[(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)methylidene]pyrrolidine-l-carboxylate Int. A (CAS No: 2376764-71-3) (858 mg, 2.777 mmol, 1.2 equiv), Pd(dtbpf)Ch (150 mg, 0.231 mmol, 0.1 equiv) and K3PO4 (1228 mg, 5.785 mmol, 2.5 equiv) in Dioxane (10 mL) and H2O (2 mL) was stirred at 100°C for Ih under nitrogen atmosphere. The resulting mixture was fdtered, the filter cake was washed with EtOAc (3x10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1: 1) to afford tert-butyl (3E)-3-{[6- (methylcarbamoyl)pyridazin-3-yl]methylidene}pyrrolidine-l-carboxylate Id (200 mg, 27.14%yield) and tert-butyl (3Z)-3-{[6-(methylcarbamoyl)pyridazin-3- yl]methylidene}pyrrolidine-l-carboxylate 1c (200 mg, 27.14%yield) as a white solid.LCMS 1c :(ES,m / z):319[M+l]+ ; LCMS Id :(ES,m / z):319[M+l]+'H NMR- Id tert-butyl (3E)-3-{ [6-(methylcarbamoyl)pyridazin-3- yl]methylidene}pyrrolidine-l-carboxylate (400 MHz, DMSO-c / e) 5 9.25 (q, IH), 8.13 (d, J = 8.7 Hz, IH), 7.77 (d, J= 8.8 Hz, IH), 6.81 - 6.69 (m, IH), 4.21 (d, J= 2.5 Hz, 2H), 3.52 (t, J = 1A Hz. 2H), 3.14 (s, 2H), 2.86 (d, J= 4.8 Hz, 3H), 1.43 (s, 9H).NOESY (400 MHz, DMSO-Je): the proton at 6.8 ppm has correlation with the proton at 4.21 ppm.'H NMR- 1c tert-butyl (3Z)-3-{[6-(methylcarbamoyl)pyridazin-3- yl]methylidene}pyrrolidine-l-carboxylate (400 MHz, DMSO-cL) 5 9.32 (q, J= 4.7 Hz, IH), 8.13 (d, J= 8.6 Hz, IH), 7.75 (d, J= 8.1 Hz, IH), 6.70 (t, J= 2.5 Hz, IH), 4.45 (d, J= 2.7 Hz. 2H), 3.44 (t, J= 7.3 Hz, 2H), 2.86 (m. 5H), 1.43 (s, 9H).NOESY- (400 MHz, DMSO-de): the proton at 6.70 ppm has correlation with the proton at 2.86 ppm.

[0118] Synthesis of le

[0119] A solution of tert-butyl (3E)-3-{[6-(methylcarbamoyl)pyridazin-3- yl]methylidene}pyrrolidine-l-carboxylate Id (200 mg, 0.628 mmol, 1 equiv) in DCM (6 mL) was added trifluoroacetic acid (2 mL). The reaction mixture was stirred at room temperature for Ih. The resulting mixture was concentrated under reduced pressure. This resulted in N- methyl-6-[(3E)-pyrrolidin-3-ylidenemethyl]pyridazine-3-carboxamide TFA salt le (200 mg) as a crude brown oil.LCMS :(ES,m / z):219[M+l]+

[0120] Synthesis of compound 1

[0121] A solution of N-methyl-6-[(3E)-pyrrolidin-3-ylidenemethyl]pyridazine-3- carboxamide TFA salt crude product le (200 mg), 7-(chloromethyl)-3-ethyl-lH-l,5- naphthyridin-2-one Int. B (136 mg, 0.611 mmol, 1 equiv), KI (20 mg, 0.122 mmol, 0.2 equiv) and DIEA (394 mg, 3.053 mmol, 5 equiv) in DMF (3 mL) was stirred at 80°C for Ih under nitrogen atmosphere. The reaction mixture was filtered and purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD Cl 8 Column 30*150 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow- rate: 60 mL / min mL / min; Gradient: 5% B to 5% B in 1 min, 5% B to 15% B in 2 min, 15% to 35% B in 11 min; Wave Length: 254nm / 220nm nm; RTl(min): 8.42) to afford 6-{[(3E)-l-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]pyrrolidin-3-ylidene]methyl}-N-methylpyridazine-3- carboxamide compound 1 (26.4 mg, 10.68%yield, 98.0%purity) as a white solid.LCMS :(ES,m / z):405[M+l]+JH NMR (400 MHz, DMSO-Je) 5 11.86 (s, 1H), 9.24 (q, J= 4.7 Hz, 1H), 8.42 (d, J= 1.9 Hz, 1H), 8.10 (d, J= 8.7 Hz, 1H), 7.79 - 7.67 (m, 2H), 7.64 (d, J= 1.8 Hz, 1H), 6.67 (t, J= 2.3 Hz, 1H), 3.78 (s, 2H), 3.42 (s, 2H), 2.97 (s, 2H), 2.89 - 2.78 (m, 3H), 2.78 - 2.64 (m, 2H), 2.61 - 2.53 (m, 2H), 1.19 (t, J= 7.4 Hz, 3H).Preparation of compound 2Compound 2 was prepared similarly as compound 1.LCMS :(ES,m / z):405 [M+l]+1H NMR-(400 MHz, DMSO-ds) 5 11.85 (s, 1H), 9.20 (q, J = 4.8 Hz, 1H), 8.44 (d, J= 1.8 Hz, 1H), 8.07 (d. J= 8.7 Hz. 1H), 7.76 (s, 1H), 7.69 (d, J = 8.7 Hz, 1H), 7.65 (d, J = 1.8 Hz, 1H), 6.61 (t, J= 2.2 Hz, 1H), 3.84 (s, 2H), 3.70 (d, J= 2.6 Hz, 2H), 2.83 - 2.77 (m, 4H), 2.75 (d, J = 5.5 Hz, 3H), 2.56 (td, J= 7.4, 1.2 Hz, 2H), 1.19 (t, J= 7.4 Hz, 3H).

[0122] Preparation of compound 3 and 4

[0123] Compound 3 and 4 were prepared similarly as compound 1 and 2 except the separation was done at the final stage. The crude product was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IA, 3*25 cm. 5 pm; Mobile Phase A: Hex( lOmM NHs-MeOH), Mobile Phase B: IP A; Flow rate: 30 mL / min; Gradient: isocratic 50; Wave Length: 220 / 254 nm; RTl(min): 17.12; RT2(min): 26.19; Sample Solvent: MeOH: DCM=1 : 1— HPLC; Injection Volume: 1.5 mL; Number Of Runs: 3) to afford 5-{[(3E)-l-[(7- ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]pyrrolidin-3-ylidene]methyl}-N- methylpyridine-2-carboxamide compound 4 (25.7 mg. 10.38%yield, 99.9%purity) as an off- white solid and 5-{[(3Z)-l-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]pyrrolidin-3- ylidene]methyl}-N-methylpyridine-2-carboxamide compound 3 (35.4 mg, 14.30%yield, 98.9%purity) as an off-white solid.LCMS-compound 3 :(ES,m / z):404[M+l]+1H NMR- compound 3 5-{[(3Z)-l-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3- yl)methyl]pyrrolidin-3-ylidene]methyl} -N-methylpyridine-2-carboxamide (400 MHz, DMS0-O 5 11.83 (s, 1H), 8.68 (d, J= 4.9 Hz, 1H), 8.43 (dd, J= 8.8, 2.0 Hz, 2H), 7.95 (d, J = 8.0 Hz. 1H), 7.73 (d. J= 7.8 Hz. 2H), 7.62 (d. J= 1.9 Hz. 1H), 6.47 (s. 1H), 3.82 (s, 2H), 3.50 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H), 2.69 (s, 4H), 2.60 - 2.52 (m, 2H), 1.18 (t, J = 7.4 Hz, 3H).1D-NOESY- compound 3 (400 MHz, DMSO-d6): the proton at 6.47 ppm has correlation with the proton at 2.70 ppm.LCMS- compound 4:(ES,m / z):404[M+l]+'H NMR- compound 4 5-{[(3E)-l-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3- yl)methyl]pyrrolidin-3-ylidene]methyl}-N-methylpyridine-2-carboxamide (400 MHz, DMS0-O 5 11.86 (s, IH), 8.70 (q, J= 4.7 Hz, IH), 8.54 (d, J= 2.1 Hz, IH), 8.41 (d, J= 1.8 Hz, IH), 7.98 (d, .7 = 8.2 Hz, IH), 7.91 (dd, J= 8.3, 2.2 Hz, IH), 7.76 (q, .7= 1 .0 Hz, IH), 7.67 - 7.57 (m, IH), 6.47 (s, IH), 3.75 (s, 2H), 3.35 (s, 3H), 2.81 (d, J= 4.8 Hz, 3H), 2.77 (s, 4H), 2.62 - 2.53 (m, 2H), 1.19 (t, J= 7.4 Hz, 3H).1D-N0ESY- compound 4 (400 MHz, DMSO-de): the proton at 6.47 ppm has correlation with the proton at 3.34 ppm.

[0124] Preparation of compound 5

[0125] To a stirred mixture of 5-bromo-A-methylpicolinamide 3a (300 mg, 1.395 mmol, 1 equiv) and tert-butyl 3-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl) methylene) azetidine- 1 -carboxylate Int. C (CAS NO: 2246802-17-3) (452 mg. 1.535 mmol, 1.1 equiv) in Dioxane (3 mL) and H2O (600 uL) were added Pd(dtbpf)C12 (90 mg, 0.140 mmol, 0.1 equiv) and KSPO4 (740 mg, 3.487 mmol, 2.5 equiv) at 25°C under argon atmosphere. The resulting mixture was stirred at 100°C for Ih. The reaction w as diluted with water (10 mL). The resulting mixture was extracted with EA (3 x lOmL). The combined organic layers were washed with brine (2x10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentratedunder reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1 : 1) to afford tert-butyl 3-((6-(methylcarbamoyl) pyridin-3-yl) methylene) azetidine-1 -carboxylate 5b (390 mg, 92. 16%yield) as a light yellow^ solid.LCMS-:(ES, m / z):304.05[M+H]+

[0126] Synthesis of 5c

[0127] To a stirred solution of tert-butyl 3-((6-(methylcarbamoyl) pyridin-3-yl) methylene) azetidine-1 -carboxylate 5b (290 mg, 0.956 mmol, 1 equiv) in DCM (3 mL) was added TFA (1 mL) at 0°C. The resulting mixture was stirred at 25°C for 2h. The resulting mixture was concentrated under reduced pressure. This resulted in 5-(azetidin-3- ylidenemethyl)-A-methylpicolinamide TFA salt 5c (300 mg) as a crude light brown solid.LCMS-:(ES.m / z):204. 10[M+H]+

[0128] Synthesis of compound 5

[0129] To a stirred mixture of 5-(azetidin-3-ylidenemethyl)-A-methylpicolinamide TFA salt crude product 5c (20 mg) and 7-(chloromethyl)-3-ethyl-l,5-naphthyridin-2(lrt)-one (24 mg, 0. 108 mmol, 1.1 equiv) in DMF (1 mL) were added KI (3 mg, 0.020 mmol, 0.2 equiv) and DIEA (127 mg, 0.980 mmol, 10 equiv) at 25°C under argon atmosphere. The resulting mixture was stirred at 80°C for Ih. The reaction mixture was filtered and purified by reversed- phase flash chromatography with the following conditions: Column: YMC Triart C18 ExRs 5pm, 30 mm * 150 mm; Mobile Phase A: Water (lOmmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient: 15% B to 38% B in 10 min; Wave Length: 254nm / 220nm nm; RTl(min): 9.52. to afford 5-((l-((7-ethyl-6-oxo-5,6-dihydro-l,5- naphthyridin-3-yl) methyl) azetidin-3-ylidene) methyl)-A-methylpicolinamide compound 5 (25.2 mg, 65.75%yield, 99.7%purity) as a white solid.LCMS:(ES, m / z):390.15 [M+H]+

[0130] 'H NMR (400 MHz, Chloroform-d) 5 10.61 (s, 1H), 8.50 (d, J = 1.8 Hz, 1H), 8.28 (d, J= 2.1 Hz, 1H), 8.12 (d, J = 8.1 Hz, 1H), 7.93 (d, J= 5.5 Hz, 1H), 7.84 (s, 1H), 7.64 (s, 1H), 7.49 (dd. J= 8.2, 2.2 Hz, 1H). 6.30 - 6.25 (m, 1H). 4.34 (s, 2H), 4.18 (s. 2H), 3.99 (s, 2H), 3.03 (d, J = 5. 1 Hz, 3H), 2.73 (qd, J = 7.4, 1.4 Hz, 2H), 1.31 (t, J = 7.4 Hz, 3H).

[0131] Preparation of compound 6

[0133] A solution of methyl 5-bromo-6-fluoropyridine-2-carboxylate 6a (1 g, 4.273 mmol, 1 equiv) and methylamine (2M in EtOH)(2.13 mL, 4.273 mmol, 1 equiv) in methanol(50 mL) was stirred at room temperature for Ih. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (5: 1) to afford 5-bromo-6-fluoro-N-methylpyridine-2-carboxamide 6b (400 mg, 40.17%yield) as a white solid. LCMS:(ES,m / z):233[M+l]+

[0134] Synthesis of 6c and 6d

[0135] A solution of 5-bromo-6-fluoro-N-methylpyridine-2-carboxamide 6b (500 mg, 2.146 mmol, 1 equiv), tert-butyl 3-[(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)methylidene]pyrrolidine-l-carboxylate(CAS No: 2376764-71-3) (796 mg, 2.575 mmol, 1.2 equiv), Pd(dtbpf)C12 (139 mg, 0.215 mmol, 0.1 equiv) and K3PO4 (1138 mg, 5.365 mmol, 2.5 equiv) in Dioxane (10 mL) and H2O (2 mL) was stirred at 100°C for Ih under nitrogen atmosphere. The resulting mixture was filtered, the filter cake was washed with EtOAc (3x10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (5: 1) to afford tert-butyl (3E)-3-{[2-fluoro- 6-(methylcarbamoyl)pyridin-3-yl]methylidene}pyrrolidine-l-carboxylate 6c (200 mg, 27.79%yield) and tert-butyl (3Z)-3-{[2-fluoro-6-(methylcarbamoyl)pyridin-3- yl]methylidene}pyrrolidine-l-carboxylate 6d (200 mg, 27.79%yield) as a white solid.LCMS-6c :(ES,m / z):336[M+l]+LCMS-6d :(ES,m / z):336[M+l]+'H NMR-6C terl-butyl (3E)-3-{[2-fluoro-6-(methylcarbamoyl)pyridin-3- yl]methylidene}pyrrolidine-l-carboxylate (400 MHz. DMSO-d6) 5 8.64 (q. J = 4.7 Hz, 1H).8.10 (dd, J = 9.8, 7.7 Hz, 1H), 7.95 (dd, J = 7.7, 1.5 Hz, 1H), 6.53 (t, 7 = 2.5 Hz, 1H), 4.13 (s, 2H), 3.46 (t, 7= 7.3 Hz, 2H), 2.80 (d, 7= 4.8 Hz, 3H), 1.42 (s, 9H).NOESY(400 MHz, DMSO-d6): the proton at 6.53 ppm has correlation with the proton at 4. 13 ppm.1H NMR-6d tert-butyl (3Z)-3-{[2-fluoro-6-(methylcarbamoyl)pyri din-3 - yl]methylidene}pyrrolidine-l-carboxylate (400 MHz, DMSO-d6) δ 8.64 (q, J= 4.8 Hz, IH), 7.97 (d, J= 3.8 Hz, 2H), 6.59 - 6.44 (m, IH), 4.12 (s, 2H), 3.38 (d, J= 7.4 Hz, 2H), 2.90 - 2.75 (m, 5H), 1.41 (s, 9H).NOESY- (400 MHz, DMSO-d6): the proton at 6.59 ppm has correlation with the proton at 2.80 ppm.

[0136] Synthesis of 6e

[0137] A solution of tert-butyl (3E)-3-{[2-fluoro-6-(methylcarbamoyl)pyridin-3- yl]methylidene}pyrrolidine-l-carboxylate 6c (200 mg, 0.596 mmol, 1 equiv) in DCM (6 mL) was added trifluoroacetic acid (2 mL). The reaction mixture was stirred at room temperature for Ih. The resulting mixture was concentrated under reduced pressure. This resulted in 6- fluoro-N-methyl-5-[(3E)-pyrrolidin-3-ylidenemethyl]pyridine-2-carboxamide TFA salt 6e (200 mg) as a brown oil.LCMS:(ES,m / z):236[M+l]+

[0138] Synthesis of compound 6

[0139] A solution of 6-fluoro-N-methyl-5-[(3E)-pyrrolidin-3-ylidenemethyl]pyridine- 2-carboxamide TFA salt crude product 6e (100 mg), 7-(chloromethyl)-8-fluoro-3-methyl-lH- quinoxalin-2-one (64 mg, 0.283 mmol, 1 equiv), KI (9 mg, 0.057 mmol, 0.2 equiv) and DIEA(183 mg, 1.417 mmol, 5 equiv) in DMF (3 mL) was stirred at 80°C for Ih under nitrogen atmosphere. The reaction mixture was filtered and purified by Prep-HPLC with the following conditions (Column: XBridge Shield RP 18 OBD Column 30*250 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 21% B to 35% B in 11 min; Wave Length: 254nm / 220 nm; RTl(min): 9.75) to afford 6-fluoro-5- {[(3E)-l-[(5-fluoro-2-methyl-3-oxo-4H-quinoxalin-6-yl)methyl]pyrrolidin-3- ylidene]methyl}-N-methylpyridine-2-carboxamide compound 6 (29.5 mg. 24.47%yield. 98.4%purity) as a white solid.LCMS:(ES,m / z):426[M+l]+'H NMR (400 MHz. DMSO-O 5 12.44 (s, IH), 8.61 (q, J= 4.8 Hz, IH), 8.15 (dd, J= 9.9, 7.8 Hz, IH), 7.92 (dd, J= 7.8, 1.5 Hz, IH), 7.51 (d, J= 8.3 Hz, IH), 7.34 - 7.25 (m, IH), 6.42 (t, J= 2.3 Hz, IH), 3.79 (s, 2H), 2.79 (d, J= 4.8 Hz, 3H), 2.76 - 2.69 (m, 2H), 2.52 (s, 2H), 2.50 (d, J= 2.1 Hz, 2H), 2.42 (s, 3H).19F NMR (377 MHz, DMSO-d6) 5 -72.206,-135.871

[0140] Preparation of compound 7

[0141] Compound 7 was prepared similarly as compound 6 using 6d.LCMS:(ES,m / z):426[M+l]+1H NMR (400 MHz, DMSO-O 512.44 (s, IH), 8.61 (d, J= 4.9 Hz, IH), 8.07 - 7.77 (m, 2H), 7.50 (d, J= 8.3 Hz, IH), 7.34 - 7.09 (m, IH), 6.39 (s, IH), 3.84 (s, 2H), 3.57 - 3.47 (m, 2H), 2.79 (d, J= 4.8 Hz, 3H), 2.68 (d, J= 2.9 Hz, 4H), 2.41 (s, 2H).19F NMR (377 MHz, DMSO-d6) 5 -72.043,-135.947

[0142] Preparation of compound 8LCMS :(ES,m / z):409[M+l]+‘H NMR (400 MHz, DMSO-O 5 12.43 (s, 1H), 8.09 (d, J= 8.7 Hz, 1H), 7.74 (d, J= 8.8 Hz, 1H), 7.52 (d, J= 8.3 Hz, 1H), 7.35 - 7.26 (m, 1H), 6.66 (t, J= 2.3 Hz, 1H), 3.90 - 3.73 (m, 2H), 3.49 - 3.39 (m, 2H), 2.95 (s, 2H), 2.85 (d, J= 4.8 Hz, 3H), 2.52 (s, 2H), 2.42 (s, 3H).19F NMR (377 MHz, DMSO-d6) 5 -135.864

[0143] Preparation of compound 9LCMS:(ES,m / z):409[M+l]+'H NMR(400 MHz, DMSO-d6) 5 12.46 (s, 1H), 9.22 (d, J= 4.9 Hz, 1H), 8.08 (d, J= 8.7 Hz, 1H), 7.69 (d. J= 8.7 Hz. 1H), 7.52 (d. J= 8.4 Hz. 1H), 7.33 (t, J= 7.7 Hz, 1H), 6.60 (s, 1H). 3.86 (s, 2H), 3.74 (s, 2H), 2.83 (d, J = 4.7 Hz, 3H), 2.78 - 2.69 (m, 4H), 2.42 (s, 3H).19F NMR (377 MHz, DMSO-d6) 5 -135.968

[0144] Preparation of compound 10LCMS :(ES, m / z):394.15[M+H]+1H NMR (400 MHz, DMSO-d6) 5 8.72 (q, J = 4.8 Hz, 1H), 8.42 (d, J = 2.1 Hz, 1H), 7.96 (d, J= 8.1 Hz, 1H), 7.67 (dd, J= 8.2, 2.2 Hz, 1H), 7.53 (d, J= 8.3 Hz, 1H), 7.32 (t, 7.7 Hz,1H), 4.29 (d. J = 4.2 Hz. 2H), 4.21 - 3.99 (m, 2H), 3.92 (s, 2H), 2.87 - 2.64 (m, 3H).19F NMR (376 MHz. DMSO-d6) 5 -135.85.

[0145] Preparation of compound 11Compound 11 was prepared similarly as compound 5.LCMS :(ES,m / z):391 [M+l]+1H NMR (400 MHz, DMSO-d6) 5 11.86 (s, 1H), 8.60 (q, J = 4.6 Hz, 1H), 8.41 (d, J= 1.9 Hz, 1H), 8.02 (d, J= 9.3 Hz, 1H), 7.85 (d, J= 1.5 Hz, 1H), 7.74 (d, J= 1.4 Hz, 1H), 7.65 (d, J= 1.8 Hz, 1H), 7.15 (d, J= 9.3 Hz, 1H), 6.65 (d, J = 1.6 Hz, 1H), 3.83 (d, J= 6.8 Hz, 4H), 2.82 (d, J= 4.8 Hz, 3H), 2.55 (td. J= 7.4, 1.2 Hz, 2H). 1.18 (t, J = 7.4 Hz. 3H).

[0146] Preparation of compound 12LCMS:(ES,m / z):395[M+l]+1H NMR (400 MHz, DMSO-d6) 5 8.60 (q, J = 4.7 Hz, 1H), 8.01 (d, J = 9.3 Hz, 1H), 7.84 (d, J= 1.6 Hz, 1H), 7.51 (dd, J= 8.4, 1.0 Hz, 1H), 7.37 (dd, J= 8.3, 7.1 Hz, 1H), 7.14 (d, J = 9.3 Hz. 1H), 6.64 (d. J= 1.6 Hz. 1H), 3.88 - 3.78 (m, 4H), 2.82 (d, J= 4.8 Hz, 3H), 2.41 (s, 3H).19F NMR (376 MHz. DMSO-d6) 5 -137.12.

[0147] Specific reaction conditions for individual steps to make compounds such as int.B and int. D are available in publications including for example WO2021 / 13735 (see alsoEuropean Journal of Organic Chemistry', 2019, vol. 2019, # 33, p. 5624 - 5635), the entire disclosure of which is hereby incorporated by reference Some of the intermediates are also commercially available.

[0148] Example A: Biochemical assay of PARP1 and PARP2Compounds of interest were tested in fluorescence polarization (FP) binding assay of PARP1 and PARP2. In this biochemical FP assay, binding of test compound to recombinant human PARP1 and PARP2 competes with the binding of fluorescent probe, and consequently reduces the fluorescence polarization signals. The PARP1 and PARP2 protein are available from BPS Biosciences, PARPi-FL probe is available from Tocris Bioscience. The assay buffer consisted of 50 mM Tris (pH 8.0), 0.001% Triton X-100, 10 mM MgCh, and 150 mM NaCl. Compounds were diluted to the highest concentration in a 384PP plate and then serially transferred to an Optiplate-384F plate. To the assay plate, 20 nL of each compound or DMSO was added, followed by 10 mb of 40 nM PARP1 or PARP2, both diluted in the assay buffer. The assay plate was centrifuged at 1000 rpm for 1 minute and incubated at room temperature for 30 minutes. Next , 10 mL of 6 nM PARPi-FL, also diluted in the assay buffer, was added to the plate, achieving final concentrations of 20 nM for PARP1 or PARP2, and 3 nM for PARPi-FL. The plate was centrifuged again at 1000 rpm for 1 minute, then incubated at room temperature for 4 hours. The plates were reading using an Envision reader with an excitation filter. Percent inhibition was calculated from millpolarization (mP) values using Inhibition (%) = [l-(mPc- mPL) / (mPH-mPL)]*100%, mPC, rnPL and mPH are the mP values of test compounds, low controls, and high controls, respectively. Binding IC50 values were calculated using XLFit (equation 201: y = (A+((B-A) / (l+((C / x)AD)))), where A = Bottom, B = Top, C = IC50, D = slope) with a floating top and bottom for curves.

[0149] Table 1. IC50 Values of Positive Controls and Test Compounds in PARP1 and PARP2 FP assays

[0150] Example B: Cellular activity testing

[0151] The survival of BRCA2 deficient cells and wild type cells in the presence of PARP-1 inhibitors were investigated with luminescent cell viability assay. Two isogenic cell lines of colorectal adenocarcinoma, DLD-1 wild-type and DLD-1 BRCA (- / -) (differing by the presence or absence of both BRCA2 allele), are utilized. Cells are harvested during their logarithmic growth phase, counted, and seeded into a 384-well plate at densities of 50 cells / well for DLD-1 wild-type and 200 cells / well for DLD-1 BRCA (- / -). After seeding, the cells are incubated overnight at 37°C with 5% CO2. The next day, cells are treated with serially diluted test compounds at ten different concentrations (ranging from 1.5 nM to 30 mM) to generate dose-response curves. Plates are incubated for additional seven days in a humidified incubator at 37°C with 5% CO2. Cell viability is assessed by measuring luminescence following the addition of Cell Titer-Go reagent (Vazyme, DD1101-02), according to the manufacturer’s protocol. IC50 values were calculated using XLFit (equation 201 : y = (A+((B- A) / (l+((C / x)AD)))), where A = Bottom, B = Top, C = IC50. D = slope) with a floating top and bottom for curves.

[0152] Table 2. EC50 Values of Positive Controls and Test Compounds in Cells

[0153] The effects of the test compounds on the viability of other cell lines, such as MDA-MB-436, MDA-MB-231. SUM149PT, HCC1395, and UWB 1.289, are determined using a similar method.

[0154] Example C: Evaluation of compound effect on efflux

[0155] Bidirectional Permeability of Compounds in MDCKIIKO-Human MDR1 Cell Line

[0156] MDCKIIKO-Human MDR1 cells were diluted to 1.56xl06cells / mL in culture medium, and 50 pL of cell suspension was dispensed into the filter well of the pre-incubated 96-well HTS Transwell plate. Cells were cultivated for 4-8 days in a cell culture incubator at 37 °C, 5% CO2, and 95% relative humidity, with the culture medium being replaced every other day.

[0157] Stock solutions (10 mM) of the test and control compounds were prepared in DMSO, with metoprolol and Digoxin serving as control compounds. Transepithelial electrical resistance (TEER) across the monolayer was measured using Millicell Epithelial Volt-Ohm measuring system (Millipore, USA) to assess the integrity' of the cell monolayer. Following TEER measurement, the plate was returned to the incubator.

[0158] Before the assay, the MDCKIIK0-Human MDR1 monolayer was washed three times with pre-warmed HBSS (10 mM HEPES, pH 7.4), then allowed to settle at 37 °C for 30 minutes. To prepare the sample solution, 3 pL of compound solution (0.2 mM in DMSO) were added to 597 pL of transport buffer to obtain a 1 pM compound working solution. The plate was shaken at 1000 rpm for 10 min to ensure adequate mixing. The final concentration of DMSO in the incubation system was 0.5%.

[0159] For determining the rate of drug transport in the apical -to-basolateral direction (A^B), 125 pL of the 1 pM working solution was added to the Transwell insert (apical compartment). A 50 pL sample was immediately taken from the apical compartment and transferred to 250 pL of quenching solvent in a new 96-well plate as the initial donor sample. The plate was shaken at 1000 rpm for 5 minutes, and 235 pL of transport buffer was added to the receiver plate (basolateral compartment). For basolateral-to-apical transport (B^A), 285 pL of the 1 pM working solution was added to the basolateral compartment. Similarly, a 50 pL sample was immediately taken from the basolateral compartment and transferred to 250 pL of quenching solvent in a new 96-well plate as the initial donor sample. The Transwell insert (apical compartment) was filled with 75 pL of transport buffer. The plate was shaken at 1000 rpm for 5 minutes.

[0160] Both A^B and B— >A sample transfers were performed concomitantly at 37°C for 2 hours without shaking. At the end of the transport period, 50 pL samples were taken from the donor sides (apical compartment for A^B flux, and basolateral compartment for B— >A flux) and transferred to 250 pL quenching solvent in new 96-well plates. Similarly, 50 pL was directly removed from the receiver sides and transferred to the quenching solvent.

[0161] The samples were vortexed at 1000 rpm for 5 minutes and centrifuged at 4000 rpm for 20 minutes. An aliquot of 100 pL of the supernatant was mixed with 100 pL of ultrapure H2O for LC / MS / MS analysis. All incubations were performed in duplicate.

[0162] Table 3. Permeability7Results of Test Compounds and Control Compounds in MDCKIIK0-Human MDR1 Cell Line

[0163] Caco-2 BCRP1 Assay. Standard Caco-2 BCRP1 (Breast Cancer Resistance Protein) assay is used to evaluate the interaction of compounds with the BCRP1 efflux transporter.

[0164] Example D: Liver microsome stability assay

[0165] A master solution containing phosphate buffer, ultra-pure H2O, and microsomes was prepared to achieve final concentrations of 100 pM for phosphate buffer and 0.5 mg / mL for microsomes. Subsequently, 80 nL of 1 mM test compounds were added to 72 pL of the master solution. The reaction was initiated by adding 8 pL of 10 mM NADPH solution, resulting in a final NADPH concentration of 1 mM. Samples containing NADPH were prepared in duplicate. Verapamil served as the positive control for this study. Both the test and control compounds were set at a final concentration of 1 pM, with the DMSO concentration maintained at 0.1% in the incubation system.

[0166] The mixture was pre-warmed at 37°C for 10 minutes. Afterward, aliquots of 10 pL from each of the four test compounds (40 pL total) were taken from the reaction solution and transferred into a new plate containing 120 pL of cold acetonitrile with internal standards (IS: 200 nM labetalol, 100 nM tolbutamide, and 100 nM ketoprofen). The same procedure was followed after 60 minutes of incubation at 37°C.

[0167] The samples were centrifuged at 3,220 g for 45 minutes, and 40 pL of the supernatant was mixed with 40 pL of ultra-pure H2O for subsequent LC-MS / MS analysis. Peak areas were determined from the extracted ion chromatograms. The slope value, k, was calculated using linear regression of the natural logarithm of the remaining parent drug percentage vs. incubation time. The in vitro half-life (ti / 2) was then determined based on the slope value.

[0168] In vitro 11 / 2= -(0.693 / C)

[0169] Conversion of the in vitro ti / 2 (min) into the in vitro intrinsic clearance (in vitro CLint, in pL / min / mg protein) was done using the following equation (mean of duplicate determinations):

[0170] In vitro CLint = (0.693 / ti / 2)* [volume of incubation (mL) / amount of proteins (mg)]

[0171] Table 4 Liver microsomes stability at 1 uM

[0172] Example E: in vivo brain exposure assay

[0173] A compound of interest in a formulation such as a suspension of 1% methylcellulose (MC) in deionized water is administered orally at a dose of 10 mg / kg to six non-fasted male BALB / c mice (6-8 weeks old, 20-25 g). Brain and blood samples are collected at 0.25, 0.5, 1, 2, 4, and 7 hours after administration. Plasma is obtained by centrifuging the blood samples at 4000g and 4°C for 5 minutes. Brain tissue is homogenized after adding four times the volume of phosphate-buffered saline (pH 7.4). The compounds in the plasma and brain are quantified using LC-MS / MS. The area under the curve (AUC) is determined for the brain tissue and plasma from 0 to 7 hours.

[0174] The total brain-to-plasma concentration ratio Kp is determined using the following formula:

[0175] Brain Kp =AUC0-7hours (brain) / AUC0-7hours (plasma)

[0176] The unbound fraction in plasma (fu, plasma) and the unbound fraction in brain (fu. brain) are obtained from in vitro equilibrium dialysis of plasma and brain homogenate, respectively. The unbound brain-to-plasma ratio is determined using the following formula:

[0177] Brain Kp. uu =Kp x fu. brain / fu, plasma

[0178] All references cited herein are incorporated by reference in their entireties.

[0179] It will be appreciated by persons skilled in the art that the invention described herein is not limited to what has been particularly shown and described. Rather, the scope of the invention is defined by the claims which follow; It should further be understood that the above description is only representative of illustrative examples of embodiments. The description has not attempted to exhaustively enumerate all possible variations. The alternate embodiments may not have been presented for a specific substituent of the compound, or a step of the method, and may result from a different combination of described substituent or step, or that other undescribed alternate embodiments may be available for a compound or method, is not to be considered a disclaimer of those alternate embodiments. It will be appreciated that many of those un-described embodiments are within the literal scope of the following claims, and others are equivalent.

Claims

CLAIMS1. A compound of formula I, a geometric isomer thereof, or a pharmaceutically acceptable salt thereof,whereinA is 5- or 6- membered aromatic ring;B is a 4-8 membered carbocyclic ring or heterocyclic ring;C is 9-10 membered bicyclic ring;L is C(O), O. SO2, C1-3alkylene or NRn;M is H, deuterium, halogen, C3-6cycloalkyl or Ci-ealkyl;Rain each instance is independently selected from the group consisting of deuterium, OC1-6alkyl, SC1-6alkyl, CN, OH, SH, halogen, NO2, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C 1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, C1-6alkyl, hydroxyC 1-6alkyl, dihydroxyC 1-10alkyl, C3-6cycloalkyl, C(=NC 1-6alkyl)C1-6alkyl, OC(O)N(Rm)2, C(O)SRm, OC1-6alkyleneOCi ealkyl, OC1-6alkyleneO-haloC 1-6alkyl, SC1- 6alkyleneOC1-6alkyl, SC1-6alkyleneSC1-6alkyl, OC1-6alkyleneSCi-ealkyl, SC1- 6alkyleneO-haloC1-6alkyl, SC1-6alkyleneS-haloC1-6alkyl. OC1-6alkyleneS-haloC1- 6alkyl, C1-6alkylene-CN, OC1-6alkylene-CN. SC1-6alkylene-CN. OC1-6alkylene- N(Rm)2, C2-6alkynyl, C2-6alkenyl, SO2N(Rm)2, NRmSO2C1-6alkyl, C1-6alkylSO2(sulfone), S(O)OH, C1-6alkylS(O) (sulfoxide), nitroso, and C1-6alkylOSO2;Rbin each instance is independently selected from the group consisting of deuterium, OC1-6alkyl. SCi-ealkyl. CN, OH, SH. halogen, N(Rm)2, C(O)ORm. C(O)N(Rm)2. C(O)C1-6alkyl, haloC1-6alkyl, C1-6alkyl, hydroxyC1-6alkyl, C2-6alkynyl, and C2- 6alkenyl;Rcin each instance is independently selected from the group consisting of deuterium, oxo, OC1-6alkyl, SC1-6alkyl, CN, OH, SH, halogen, NO2, N(Rm)2, C(O)ORm, C(O)N(Rm)2. C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, C1-6alkyl. hydroxyC1- 6alkyl, dihydroxyC 1-10alkyl, C3-6cycloalkyl, C(=NC1-6alkyl)Ci-ealkyl, OC(O)N(Rm)2, C(O)SRm, OC1-6alkyleneOC1-6alkyl, OC1-6alkyleneO-haloC1-6alkyl, SC1-6alkyleneOC1-6alkyl, SC1-6alkyleneSC1-6alkyl, OC1-6alkyleneSC1-6alkyl, SC1- 6alkyleneO-haloC1-6alkyl, SC1-6alkyleneS-haloC1-6alkyl. OC1-6alkyleneS-haloC1- 6alkyl, C1-6alkylene-CN, OC1-6alkylene-CN, SC1-6alkylene-CN, OC1-6alkylene- N(Rm)2, C2-6alkynyl, C2-6alkenyl, SO2N(Rm)2, NRmSO2C1-6alkyl, C1-6alkylSO2(sulfone), S(O)OH, C1-6alkylS(O) (sulfoxide), nitroso, and C1-6alkylOSO2;Rmeach is independently hydrogen or C1-6alkyl or halo-C1-6alkyl;Rnis hy drogen. C1-6alkyl, halo-C 1-6alkyl, C(O)C1-6alkyl; m is 0, 1, 2, 3, 4 or 5; n is 0, 1, 2, or 3; and p is 0, 1, 2, 3, 4 or 5.

2. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of claim 1, geometric isomer thereof, or pharmaceutically acceptable salt thereof of claim 1, wherein A is an optionally substituted pyridinyl or pyrimidinyl.

3. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-2, wherein A is pyridinyl optionally substituted with one or more Ras selected from the group consisting of OC 1-6alkyl. SC1-6alkyl. CN, OH, SH, halogen, NO2, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1- 6alkyl, haloC1-6alkyleneO, C1-6alkyl, hydroxyC1-6alkyl, and dihydroxyC1-10alkyl.

4. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-3, wherein the compound is represented by Formula I-aWherein Rain each instance is independently selected from the group consisting of OCi-galkyl, CN, halogen. N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1- 6alkyl, haloC1-6alkyleneO, C1-6alkyl, and hydroxyC1-6alkyl; m is 1, 2 or 3.

5. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-4. wherein at least one of (Ra)m is C(O)NHC1-6alkyl.

6. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-5. wherein B is a 4-6 membered heterocyclic ring.

7. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-6, wherein the compound is represented by Formula I- b,wherein x is 1 or 2; and y is 1 or 2.

8. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-7, wherein x is 1, y is 1 or 2, n is 0 or 1.

9. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1 -8, wherein C comprises at least 1 ring nitrogen, and at least one ring carbon is substituted with ox.

10. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-9. wherein Rcin each instance is independently selected from the group consisting of oxo, OC 1-6alkyl, SC1-6alkyl, CN, halogen, OH, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1-6alkyleneO, and C1-6alkyl; andP is 1, 2 or 3.

11. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-10, wherein the compound is represented by Formula I- c.Wherein Rcin each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1- 6alkyleneO, and C1-6alkyl; and p is 1, 2 or 3.

12. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-10, wherein the compound is represented by Formula I- d.Wherein Rcin each instance is independently selected from the group consisting of OC1-6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1- 6alkyleneO, and C1-6alkyl; and p is 1, 2 or 3.

13. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-10, wherein the compound is represented by I-e.WhereinP is nitrogen or optionally substituted ring carbon;Q is nitrogen or optionally substituted ring carbon; and U is N or optionally substituted ring carbon;.

14. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-10, wherein the compound is represented by I-f,WhereinP is nitrogen or optionally substituted ring carbon;Q is nitrogen or optionally substituted ring carbon;T is N or optionally substituted ring carbon;U is N or optionally substituted ring carbon;Rclin each instance is independently selected from the group consisting of OC1- 6alkyl, halogen, C(O)ORm. C(O)N(Rm)2. C(O)C1-6alkyl. haloC1-6alkyl. haloC1- 6alkyleneO, and C1-6alkyl;Rc2in each instance is independently selected from the group consisting of OC1- 6alkyl, halogen, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1- 6alkyleneO, and C1-6alkyl;Rain each instance is independently selected from the group consisting of OC1-6alkyl, CN, halogen, N(Rm)2, C(O)ORm, C(O)N(Rm)2, C(O)C1-6alkyl, haloC1-6alkyl, haloC1- 6alkyleneO, C1-6alkyl, and hydroxyC1-6alkyl;Ra’ is C(O)ORm, C(O)N(Rm)2, or C(O)C1-6alkyl; m is 1, 2, or 3. wherein Rcin each instance is independently selected from the group consisting of halogen, haloC1-6alkyl, and C1-6alkyl.

15. The compound, the geometric isomer thereof, or the pharmaceutically acceptable salt thereof of any one of claims 1-14, wherein M is H.

16. The compound of formula I, geometric isomer thereof, or pharmaceutically acceptable salt thereof of claim 1. wherein the compound is selected from the group consisting of17. A pharmaceutical composition comprising a therapeutically effective amount of the compound of formula 1, geometric isomer thereof, or pharmaceutically acceptable salt thereof of any one of claims 1-16, and a pharmaceutically acceptable carrier.

18. A method of treating a disease associated with abnormal poly(ADP-ribose) polymerase (PARP) enzyme, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of formula I, geometric isomer thereof, or pharmaceutically acceptable salt thereof of any one of claims 1-16 or the pharmaceutical composition of claim 17, wherein the disease is selected from the group consisting of neurodegenerative diseases, metabolic disease, cardiovascular diseases, autoimmune disease, and cancer.

19. The method of claim 18, wherein the disease is selected from the group consisting of bladder cancer, breast cancer, cancer of the fallopian tube(s), cholagiocarcinoma, colon adenocarcinoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, gastric cancer, kidney clear cell cancer, lung cancer, mesothelioma, ovarian cancer, pancreatic cancer, peritoneal cancer, prostate cancer, uterine endometrial cancer, and uveal melanoma.

20. A method of inhibiting PARP1 enzyme, comprising contacting a cell with an effective amount of the compound of formula (I) or the pharmaceutically acceptable salt thereof of any one of claims 1-16 or the pharmaceutical composition of claim 17.

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