Tricyclic compound for breast and ovarian cancer treatment

Abstract

The present invention relates to a tricyclic compound of formula (I) and its analogue useful for the treatment of breast and ovarian cancer. In particular, the present invention relates to novel tricyclic compound as PARP1 / 2 inhibitor, designed to block the activity of the enzyme Poly (ADP-ribose) polymerase (PARP1 / 2), which plays a crucial role in DNA damage repair, particularly in cancer cells. The synthesized inhibitor molecules having improved solubility, bioavailability and no haematological toxicity can be useful in potential cancer therapies, and defects in DNA repair pathways.

Description

[0001] P_W0100786

[0002] TRICYCLIC COMPOUND FOR BREAST AND OVARIAN CANCER TREATMENT FIELD OF THE INVENTION

[0003] The present invention relates to tricyclic compound of formula (I) and its pharmaceutically acceptable salts thereof as inhibitors of PARP 1 and 2 for the treatment of breast and ovarian cancer.

[0004] BACKGROUND OF THE INVENTION PARP1 and 2 inhibitors are considered potent antitumor agents due to the fact that they act as chemo- and radiosensitizers in the conventional therapy of malignant tumours. In addition, PARP1 and 2 inhibitors can be used as effective drugs against tumours with broken DNA repair mechanisms. Poly ADP-ribose polymerase (PARP) is a nuclear protein family consisting of 18 known members. Among the various PARP isoforms, PARP1 and PARP2 are primarily involved in single strand DNA break repair (ssDBR) processes, while other isoforms are involved in different cellular processes viz. chromatin remodelling, transcription, recombination, hypoxic response, angiogenesis, EMT, cell proliferation and programmed cell death. PARP1 and PARP2 uses NAD+ as substrate to elongate poly ADP-ribose (PAR) chain to any nucleic acid. When DNA conceives single strand break, PARP1 and PARP2 are constitutively activated and binds to the damaged DNA strand, initiates synthesizing PAR chain and transfers them to the acceptor proteins. It also recruits other DNA damage response (DDR) enzymes to the damage site, thereby facilitating the DDR response. Structurally, PARP family proteins constitute three major domains; the catalytic domain, the auto-modification domain and the DNA binding domain. In cancer cells, PARP1 and PARP2remain constitutively activated to perform extensive DNA repair activity in response to internal oxidative stress / oncogenic stimuli. This condition results in rescue from replication stalling and continuation of the cell cycle, thereby, rendering upon the cancer cells the ability to divide indefinitely. Therefore, inhibition of PARP1 and 2activations by small molecule inhibitors will be promising to stall error-prone DNA repair systems as well as to bar cancer cell divisions. Several PARP inhibitors, including olaparib, rucaparib, niraparib, and Talazoparib, are currently available on the market for Breast and Ovarian Cancer Treatment. However, their non-selective inhibition of other PARP isoforms often results in off-target effects. These off-target effects can disrupt crucial cellular processes, potentially leading to a spectrum of adverse effects experienced by patients. These may include hematologic complications such as gastrointestinal disturbances, cardiovascular issues, fatigue, and respiratory complications. P_W0100786

[0005] Furthermore, the lack of selectivity complicates the establishment of an optimal therapeutic window, a crucial concept in drug development that defines the range of doses ensuring efficacy while maintaining safety. The non-selective nature of PARP inhibitors presents challenges in achieving this balance between therapeutic effectiveness and patient safety. Consequently, current research endeavors are focused on enhancing the selectivity of PARP inhibitors. The objective is to minimize off-target effects and haematological toxicity, thereby improve the overall safety and efficacy profile of these agents for breast and ovarian cancers treatment.

[0006] In the article titled " A novel nor sesquiterpene alkaloid from the mushroom-forming fungus Flammulinavelutipes" published in Chinese Chemical Letters 2013,24, 57-58, Kai-Shun Bi et al. isolated a novel tricyclic compound A from solid cultures of Flammulinavelutipes mushroom fermented on rice. According to the study, this compound exhibited cytotoxicity against KB cells in vitro, demonstrating an IC50value of 16.6 μmol / L.

[0007] Anticancer agent

[0008]

[0009] (C)

[0010] Furthermore, the literature describes other compounds known for inhibiting PARP-I (poly (ADP-ribose) polymerase- 1). M. Tao et al., in their article titled " Synthesis and structureactivity relationships of PARP inhibitors," identified compound B as a PARP-1 inhibitor in Bioorg. Med. Chem. Lett. 2006, 16, 938-942. Additionally, several related compounds based on the structure of compound B are disclosed in a patent publication, US 2006 / 0276497A1. In the patent US9771325 published by the inventors in 2017 with tricyclic compound C is used as an anticancer agent. Where Ri and R2 are independently selected from H, alkyl, aryl, aralkyl, hydroxyalkyl, alkoxy alkyl, and nitro.

[0011]

[0012] P_W0100786

[0013] In the current approach, one of the potent PARP1 / 2 inhibitors identified by Christina et al (Clinical Cancer Research, 2010) is structure D. It is a PARP1 / 2 antagonist, administered via oral route. However, owing to its low bioavailability and poor solubility, it couldn’t pass through Phase II clinical trials. We propose generation of new analogues of this molecule while keeping intact all the functional groups required for PARP1 / 2 binding / inhibition, with the aim of achieving better PARP 1 & 2 inhibition, solubility and bio-availability with no haematological toxicity. The compounds A, B, C, and D mentioned above exhibit PARP 1 / 2 inhibitory activity, but their efficacy in the treatment of breast and ovarian cancer is not yet known.

[0014] Despite the continuous development of new anticancer agents, many of them have not proven therapeutically useful due to their low tumor selectivity and severe side effects. Therefore, there is a pressing need to discover novel potent agent for the treatment of breast and ovarian Cancer. The present invention is thus directed to meet the above requirements by synthesizing tricyclic compounds of formula I and its analogues as potent agents for Breast and Ovarian Cancer Treatment.

[0015] OBJECTIVES OF THE INVENTION

[0016] The main objective of the present invention is to provide tricyclic compound of formula (I) and its pharmaceutically acceptable salts thereof as inhibitor of poly (ADP-ribose) polymerases (PARP- 1 / P ARP-2) useful for the treatment of breast and ovarian cancer.

[0017] HN

[0018]

[0019] General Formula (I)

[0020] Wherein

[0021] n is 0,1,2...6.

[0022] R is independently selected from the group consisting of H, or 3 to 6 membered homo or hetero cycloalkyl, each of which is with or without substitution by halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. or 5 to 6 membered mono- or bicyclic homo or hetero aromatic rings wherein said hetero aromatic ring contains one or two heteroatoms selected from the group consisting of nitrogen, oxygen, and Sulphur, each of which is with or without substitution P_W0100786

[0023] halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. and each of which homo aromatics with or without substitution by halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me.

[0024] Another objective of the present invention is to validate the efficacy of the synthesized inhibitor against poly (ADP-ribose) polymerases (PARPs) through rigorous in vitro and in vivo testing. Still another objective of the invention is to synthesize tricyclic compound of formula I and its pharmaceutically acceptable salts thereof having no haematological toxicity.

[0025] Yet another objective of the present invention is to provide tricyclic compounds of formula (I) and its pharmaceutically acceptable salts with better solubility, bio-availability, better activity and no haematological toxicity thereof, useful in Breast and Ovarian Cancer treatment.

[0026] SUMMARY OF THE INVENTION

[0027] Accordingly, the present invention relates to a tricyclic compound of formula (I) and its pharmaceutically acceptable salts thereof as PARPpoly (ADP-ribose) polymerases (PARP-1 / P ARP-2) inhibitor useful in treating breast and ovarian cancer.

[0028]

[0029] General Formula (I)

[0030] Wherein

[0031] n is 0,1,2...6.

[0032] R is independently selected from the group consisting of H, or 3 to 6 membered homo or hetero cycloalkyl, each of which is with or without substitution by halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. or 5 to 6 membered mono- or bicyclic homo or hetero aromatic rings.

[0033] Wherein, the said hetero aromatic ring contains one or two hetero-atoms selected from the group consisting of nitrogen, oxygen, and Sulfur, each of which is with or without substitution halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. and each of which homo aromatics is with or without substitution by halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. P_W0100786

[0034] In a preferred embodiment, the present invention provides a tricyclic compound (formula I) and its pharmaceutically acceptable salts, useful for the treatment of Breast and Ovarian Cancer.

[0035] In another embodiment, the present invention provides a process for the synthesis of tricyclic compound of formula I for PARP poly (ADP-ribose) polymerases (PARP-1 / PARP-2) inhibition, wherein the process comprising the steps of:

[0036] i. adding (1.2 equivalents) Vinyl magnesium bromide to the solution of cyclopentane (compound 1) and diethyl ether at a temperature in the range of 0-5°C for a time period in the range of 1-1.5 hours, and quenching the reaction using NH4Cl solution followed by extracting with diethyl ether;

[0037] ii. washing and concentrating the extracted reaction mixture of step (i) to obtain a crude product. Then dissolving the obtained crude product in n-pentane and p- Toulene sulfonic acid P-TSA, (1.2 equivalents) under nitrogenic conditions at a temperature in the range of 0-5 °C for a time period of 1-1.5 hrs;

[0038] iii. quenching the reaction of step (ii) using saturated NaHCO3solution and extracting with n-pentane. Washing and drying the organic layer with anhydrous sodium sulphate, and further concentrating in water bath at a temperature in the range of 0- 15 °C to obtain compound 2 (1-vinylcyclopent-1-ene).

[0039] iv. adding Dimethyl acetylenedicarboxylate(DMAD 1.2 equivalents) to the solution of compound 2 as obtained in step (iii) dissolved in benzene and mixing it for a time period in the range of 10-12 hours to obtain a reaction mixture;

[0040] v. adding 2,3-Dichloro-5,6-dicyano-l,4-benzoquinone (DDQ 1.2 equivalents) and benzene to the reaction mixture obtained in step (iv) at room temperature and mixing it for a time period in the range of 12-14 hrs. Cooling the reaction mixture and purifying using column chromatography (15 to 25% ethyl acetate in hexane as an eluent), to obtain compound 3 (dimethyl 2,3-dihydro-1H-indene-4,5- dicarboxylate).

[0041] vi. adding NaOH (2.5 equivalents) to the solution of compound 3 (dimethyl 2,3- dihydro-1H-indene-4,5-dicarboxylate) as obtained in step (v), dissolved in THF: MeOH: H2O (3:2: 1) to form a mixture and stirring the mixture at room temperature for a time period of 5 to 7 hours, and concentrating the reaction mixture; vii. adding water and concentrated HC1 to the mixture of step (vi) to form a slurry and filtering, drying the mixture slurry to obtain crude product. Adding urea (1.2 equivalents) to the obtained crude product and heating at a temperature in the range P_W0100786

[0042] of 150°C TO 170°C for a time period of 2-4 hours to obtain compound 4 (7,8- dihy drocy clopenta [e] isoindole- 1, 3 (2H, 6H) -dione).

[0043] viii. adding a solution of nitrating mixture (1:1 H2SO4) to the crude compound 4 ((7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) obtained in step (vii) to form a reaction mixture and stirring it for a time period of 2 to 4 Hours with TLC monitoring using 30% ethyl acetate in hexane;

[0044] ix. adding the reaction mixture as obtained in step (viii) slowly to ice, resulting in a solid compound. Then filtering, drying and purifying the solid compound using column chromatography to obtain Compound 5 (5-nitro-7,8- dihy drocy clopenta [e] isoindole- 1, 3 (2H, 6H) -dione).

[0045] x. adding Pd / C (10 mol%) to the solution of compound 5 (5-nitro-7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) as obtained in step (ix) dissolved in DCM dichloromethane under a hydrogen atmosphere at room temperature for a time period in the range of 4 to 6 Hours. Then filtering and purifying the reaction mixture using column chromatography to obtain Compound 6 (5-amino-7,8- dihy drocy clopenta [e] isoindole- 1, 3 (2H, 6H) -dione).

[0046] xi. adding DMF Dimethylformamide to the compound 6 as obtained in step (x) under argon atmosphere to make a solution. Then adding l-(bromomethyl)-4- methoxybenzene and potassium iodide to the obtained solution and mixing with continuous stirring at a temperature in the range of 90 °C to 120°Cfor a time period of 12 to 16 hours to remove solid product;

[0047] xii. filtering the solution obtained in step (xi) by adding ice water and crystallizing the filtered solution by adding a mixture of dichloromethane, methanol and hexane to obtain the compounds 7-41.

[0048] In yet another embodiment, the present invention provides rigorous validation of the synthesized inhibitors against PARPs through extensive in vitro and in vivo testing.

[0049] Yet another embodiment of the present invention is to provide the novel tricyclic molecules within formula I that exhibits better activity, solubility and bio-availability with no hematological toxicity.

[0050] In another embodiment of the present invention is to provide a pharmaceutical composition for treating breast and ovarian cancer comprising of: P_W0100786

[0051] a) a compound selected from the compounds 7 to 16;

[0052] b) a pharmaceutically acceptable salt thereof.

[0053] Yet another embodiment of the present invention is to provide a method of treating breast cancer by administering the pharmaceutical composition at a particular dosage.

[0054] Still another embodiment of the present invention is to provide the enhanced efficacy and safety profile of the synthesized compound highlighting their potential as targeted therapies in oncology by leveraging selective PARP inhibition.

[0055] BRIEF DESCRIPTION OF DRAWINGS:

[0056] FIG.l: represents the PARP1 inhibitory activity of Compound 15

[0057] FIG.2: represents the cytotoxic activity of Compound 15 in DLD-1 WT (a), DLD-1 BRCA2 null (b) and in UWB 1.289 Cells (c).

[0058] FIG.3: shows the PARylation inhibition activity; Immunocytochemistry of UWB 1.289+B RCA 1 cells treated with CEP8933(10μM) and Compound 15(10μM) for 24h. Cells were stained for PAR (green), DAPI (blue) and merged under a fluorescent microscope. H2O2 at 10mM / L were taken as positive control (a) Immunoblotting analysis of PARylation inhibition activity of Compound 15 (b).

[0059] FIG.4: represents the Immunocytochemistry of UWB 1.289 cells treated with CEP8933 (1μM) and Test Compound. Cells were stained for yH2A. X (Red), DAPI (blue), and merged under a fluorescent microscope

[0060] DETAILED DESCRIPTION OF THE INVENTION:

[0061] The following description provides detailed insights into the invention, covering preferred and optional embodiments to enhance understanding and appreciation of its various aspects.

[0062] The present invention relates to a tricyclic compound of formula (I) and its pharmaceutically acceptable salts thereof as PARPpoly (ADP-ribose) polymerases (PARP- 1 / P ARP-2) inhibitor useful for the treatment of breast and ovarian cancer. P_W0100786

[0063] In an embodiment of the invention, the invention provides a tricyclic compound represented by formula I, and its pharmaceutically acceptable salts to inhibit poly (ADP-ribose) polymerases (PARP-1 / PARP-2) for the potential treatment of Breast and Ovarian Cancer.

[0064] In yet another embodiment, the present invention provides a process for synthesizing tricyclic compound of formula I and its pharmaceutically acceptable salts to enhance their solubility and bioavailability while minimizing haematological toxicity. These compounds shows promising efficacy and safety profiles as targeted therapies in oncology, through selective PARP inhibition to address critical needs in Breast and Ovarian Cancer treatment.

[0065] Yet another embodiment of the present invention is to provide a novel tricyclic compound of formula (I).

[0066] HN

[0067]

[0068] General Formula (I)

[0069] Wherein

[0070] n is 0,1,2...6.

[0071] R is independently selected from the group consisting of H, or 3 to 6 membered homo or hetero cycloalkyl, each of which is with or without substitution by halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. or 5 to 6 membered mono- or bicyclic homo or hetero aromatic rings wherein said hetero aromatic ring contains one or two hetero-atoms selected from the group consisting of nitrogen, oxygen, and Sulfur, each of which is with or without substitution halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. and each of which homo aromatics with or without substitution by halogen, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me.

[0072] Further, wherein R is independently selected from the group comprising of 3 to 6 membered homo or hetero cycloalkyl but not limited to substituted or unsubstituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, Tetrahydrofuranyl, Pyrrolidinyl, Tetrahydrothiophenyl, piperidyl, Tetrahydropyranyl, Tetrahydrothiopyranyl and 5 to 6 membered homo or hetero aromatic cyclic which are not limited to aryl, pyridinyl, furanyl, P_W0100786

[0073] pyrrolyl, pyrazolyl, imidazolyl, triazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, thiophenyl, thiazolyl, oxazolyl, napthyl.

[0074] Accordingly, the present invention encompasses the following exemplary compounds of formula I.

[0075]

[0076] In yet another embodiment, the invention provides 7-41 compounds as set forth below and its acceptable pharmaceutical salts thereof

[0077] a) 5-((4-methoxybenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (7)

[0078] b) 5-((4-(trifluoromethyl)phenethyl)amino)-7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (8)

[0079] c) 5-((3,5-bis(trifluoromethyl)benzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (9) P_W0100786

[0080] d) 5-((4-nitrobenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (10) e) 5-((2,4,6-trimethylbenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (11)

[0081] f) 5-((3,5-dinitrobenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (12)

[0082] g) 5-((3-fluoro-4-nitrobenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (13)

[0083] h) 5-((4-(trifluoromethyl)benzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (14)

[0084] i) 5-((pyridin-4-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (15)

[0085] j) 5-((4-(methylthio)benzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (16)

[0086] k) 5-((furan-2-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (17)

[0087] l) 5-(((1H-pyrrol-2-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (18)

[0088] m) 5-(((1H-pyrazol-5-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (19)

[0089] n) 5-(((1H-imidazol-5-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (20)

[0090] o) 5-(((1H-1,2,3-triazol-5-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (21)

[0091] p) 5-(thiophen-2-ylamino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (22) q) 5-((thiazol-5-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (23)

[0092] r) 5-((furan-2-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (24)

[0093] s) 5-((pyrrolidin-3 -ylmethyl)amino)-7, 8-dihydrocyclopenta[e] isoindole- 1,3 (2H,6H)- dione (25)

[0094] t) 5-((pyrimidin-4-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (26)

[0095] u) 5-((pyridazin-4-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (27) P_W0100786

[0096] v) 5-((thiophen-2-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (28)

[0097] w) 5-((piperidin-4-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (29)

[0098] x) 5-(((4-methyl-1H-pyrrol-2-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (30)

[0099] y) 5-(((2,6-dimethylpyridin-4-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (31)

[0100] z) 5-((5-chlorothiophen-2-yl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)- dione (32)

[0101] aa) 5-(((3-(trifluoromethyl)- 1H-pyrazol-5-yl)methyl)amino)-7,8- dihy drocy clopenta [e] isoindole- 1, 3 (2H, 6H) -dione (33)

[0102] bb) 5-(((2-chlorothiazol-5-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (34)

[0103] cc) 5-(((2-(trifluoromethyl)pyridin-4-yl)methyl)amino)-7,8- dihy drocy clopenta [e] isoindole- 1, 3 (2H, 6H) -dione (35)

[0104] dd) 5-(((2-fluoropyridin-4-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (36)

[0105] ee) 5-(((2-chloropyridin-4-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (37)

[0106] ff) 5-(((6-methylpyrimidin-4-yl)methyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (38)

[0107] gg) 4-(((l, 3-dioxo- 1,2, 3,6,7, 8-hexahydrocyclopenta[e]isoindol-5- yl)amino)methyl)picolinonitrile (39)

[0108] hh) 5-(((l-methyl-3-(trifluoromethyl)-1H-pyrazol-5-yl)methyl)amino)-7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (40)

[0109] ii) 5-((5-phenylpentyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (41)

[0110] In yet another embodiment of the invention, the invention provides a method for inhibiting PARP enzyme activity, by exposing the enzyme to a sufficient quantity of a compound represented by formula (I).

[0111] PARP stands for Poly (ADP-ribose): Poly: Refers to the polymerization of multiple units. ADP-ribose: ADP (Adenosine Diphosphate) is a molecule that can form chains with ribose, a type of sugar. In the context of PARP, ADP-ribose units are linked together to create a polymer. P_W0100786

[0112] Poly (ADP-ribose) Polymerase (PARP): Polymerase: This term is commonly associated with enzymes that facilitate the polymerization of molecules. In the case of PARP, it catalyses the polymerization of ADP-ribose units.

[0113] In yet another embodiment of the invention, the analogues of tricyclic compound of general formula I focuses on PARP 1 and PARP 2 inhibitory activity, solubility and bio-availability. BRCA1 (Breast Cancer gene 1) and BRCA2 (Breast Cancer gene 2) are human genes responsible for producing proteins that regulate and repair DNA.

[0114] In yet another embodiment of the invention, compound with general formula I focuses on the PARP (poly ADP-ribose polymerase) family of enzymes, which includes several isoforms. Isoforms are variations of a gene that produce slightly different forms of a protein.

[0115] In yet another embodiment of the invention is a representative structure for PARP- 1 / P ARP-2 Inhibition, Formula I.

[0116] HN

[0117]

[0118] General Formula (I)

[0119] Wherein

[0120] n is 0,1,2...6.

[0121] R is selected from the group consisting of hydrogen, homo or hetero cycloalkyl, optionally substituted withhalogen group comprising of-CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me, or 3 to 6 membered homo or hetero cycloalkyl, or 5 to 6 membered mono- or bicyclic homo or hetero aromatic rings wherein said hetero aromatic ring comprising heteroatoms is selected from the group consisting of nitrogen, oxygen, Sulfur optionally substituted with halogen group comprising of, -CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me.

[0122] Further, R independently selected from is 3 to 6 membered homo or hetero cycloalkylis not limited to substituted or unsubstituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, Tetrahydrofuranyl, Pyrrolidinyl, Tetrahydrothiophenyl, piperidyl, Tetrahydropyranyl, Tetrahydrothiopyranyl and the 5 to 6 membered homo or hetero aromatic P_W0100786

[0123] cyclics but not limited to aryl, pyridinyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, thiophenyl, thiazolyl, oxazolyl.

[0124] Still another aspect of the invention, whereinthe compound of Formula (I) is prepared fromknown starting materials, and the method for making the compounds of Formula (I) are set forth in the Examples given.

[0125] Yet anotheraspect of the present invention is to providethe PARP-1 inhibitory activity of compounds of general Formula 1.

[0126] EXAMPLES

[0127] The following examples are given by way of illustration only and therefore should not be construed to limit the scope of the present invention in any manner.

[0128] High throughput screening against PARP1:

[0129] To evaluate the PARP-1 inhibitory activity compounds, an ELISA based colorimetric assay kit (R& D Systems (Cat# 4677-096-K-104)) was used. It detects biotinylated poly (ADP-ribose) deposited by PARP-1 onto immobilized histones. The addition of Strep-HRP (Biotin-binding protein) and a colorimetric HRP substrate yields relative absorbance that correlates with PARP-1 activity.

[0130] Methodology:

[0131] To initiate the ribosylation reaction, histones were rehydrated using IX PARP buffer at room temperature. Following rehydration, test compounds were introduced along with PARP enzyme and activated DNA into the prepared wells. Detection of the reaction employed an HRP substrate, subsequently visualized using TACS -sapphire. Absorbance readings were taken at 450nm. Negative control consisted of PARP buffer alone, while positive control included activated DNA with PARP1 enzyme.

[0132] Preliminary Results:

[0133] S. No. Compound PARP1 inhibition at 100 nM

[0134] 1 Compound 7 48.34%

[0135] 2 Compound 8 41.04%

[0136] 3 Compound 9 38.6%

[0137] 4 Compound 10 25.3%

[0138] 5 Compound 11 27.45%

[0139] 6 Compound 12 27.6%

[0140] 7 Compound 13 49.3%

[0141]

[0142] P_W0100786

[0143] 8 Compound 14 25.9%

[0144] 9 Compound 15 71.6%

[0145] 10 Compound 16 8.9%

[0146]

[0147] Cell-based screening: The cell viability was determined by standard MTT dye uptake method. Briefly, DLD-1 BRCA Wild type (Catalogue number- CCL-221, (ATCC), DLD-1 BRCA2 null(Catalogue number- HD-105-007, (Horizon Discovery), UWB 1.289 (Catalogue number- CRL-2945, (ATCC) and UWB 1.289+BRC Al (Catalogue number- CRL-2946, (ATCC)Cells (3x103 cells / well) were plated into a 96 well tissue culture plate and treated with different concentration of test compounds (Compound 7 - Compound 20) in triplicate so that the final concentration of DMSO solvent was 0.2%. After 72h incubation, media and treatment was changed and cells were incubated for further 72 hrs. Same day when incubation period was completed, MTT solution was added and cells were cultured for another 4 h at 37°C in 5.0% CO2 incubator. The amount of coloured formazan derivative was determined by measuring optical density (OD) using TECAN microplate reader (Infinite M200 PRO) at 570 nm.

[0148] PARylation inhibition activity

[0149] Immunocytochemistry: UWB 1.289+BRC Al cells were plated in 48-wells plate at a seeding density of 4 × 103cells per well. Attached cells were treated with different concentrations of Compound 15 or vehicle DMSO for 24 h. H2O2 (lOmM / L) added five minutes before termination. After incubation, cells were washed with phosphate-buffered saline (PBS; Sigma, P3813) and fixed in 4% paraformaldehyde for 15 min at room temperature, thenpermeabilized with 0.1% Triton X-100 (Sigma, T8787) in PBS for 10 min and successively blocked with 1% BSA (Sigma, A9647) in PBS for 1 h. For detection of PAR, the cells were incubated with rabbit anti-PAR primary antibody (1:1000 dilution in blocking buffer) for overnight at 4°C and successively washed 3 times following incubation with Alexa Fluor 555 conjugated goat antirabbit secondary' antibody (Invitrogen, A21430) and analysed with fluorescence microscope and images were captured.

[0150] Immunoblotting: UWBL289+BRCA1 cells (0.5 x 106 cells) were plated overnight at 37°C, 5% CO2 and next day exposed to different concentrations of Compound 15 and single concentration of CEP8933 along with DMSO vehicle for 24h. Before harvesting the cells H2O2isadded at the concentration of lOmM / L for 30 minutes. Cells were accordingly harvested, extensively washed with chilled PBS, and lysed with lysis buffer (HEPES 1 mM / L, KC1 60 mM / L, NP-40 [MP Biomedicals, RIST1315] 0.3%, EDTA 1 mM / L, DTT 1 mM / L, sodium ortho vanadate 1 mM / L, PMSF 0.1 mM / L,, protease inhibitor cocktail). The cell extractions P_W0100786

[0151] were centrifuged at 13000 g for 10 min at 4°C. Protein concentration was determined by the standard Bradford method. Equal amount (25 pg) of protein from each sample was subjected to SDS-PAGE and proteins were transferred to PVDF membrane (Millipore, IPVH00010), blocked with 5% (w / v) BSA, probed with the relevant antibodies (1: 1000 dilution) for overnight at 4 °C, subsequently washed and probed with species-specific secondary antibodies coupled to horseradish peroxidase.

[0152] Y-H2A. X Foci: UWB 1.289 (ATCC) cells were seeded in 48-well plates (4 x 103cells / well) and treated with test compounds or DMSO for 72 h. Cells were fixed with 4% paraformaldehyde, permeabilized (0.1% Triton X-100), and blocked (1% BSA). yH2A. X foci were detected using rabbit anti-vH2A. X antibody (1: 1000, overnight, 4°C), followed by Alexa Fluor 555 goat anti-rabbit secondary antibody. Fluorescence images were captured by microscopy.

[0153] Table-2

[0154] Compound DLD-1 BRCA DLD-1 BRCA2 UWB1.289 BRCA1 code WT null null IC50(μM) IC50(μM) IC50(μM) Compound 7 >10 >10 >10 Compound 8 >10 >10 >10 Compound 9 >10 >10 >10 Compound 10 >10 >10 >10 Compound 11 >10 >10 >10 Compound 12 >10 >10 >10 Compound 13 >10 >10 >10 Compound 14 >10 >10 >10 Compound 15 >10 2.26 0.461 Compound 16 >10 >10 >10

[0155]

[0156] Method of preparation of tricyclic compounds:

[0157] Examples P_W0100786

[0158]

[0159] 1-vinylcyclopent-l-ene (2):

[0160] To a stirred solution of cyclopentanone (Compound 1) and diethyl ether, Vinyl magnesium bromide (1.2 equivalents) is added at a temperature 0-5°C for 1.5 hours with continuous stirring and TLC monitoring using 10% ethyl acetate in hexane. After reaction completion, the reaction is quenched with saturated NH4Cl solution and extracted with diethyl ether followed by organic layer washing with water and brine, and further drying with anhydrous sodium sulphate, then concentrating the solution under reduced pressure in water bath below 20°C temperature to obtain the crude product.

[0161] The Crude product is then dissolved in n-pentane under nitrogenic atmosphere, and p-Toluene sulfonic acid (P-TSA, 1.2 equivalents) is added to the crude productin portions at 0-5°C, with continuous stirring of the reaction mixture at the same temperature for 1 hour. Then the mixture is TLC monitored using 20% ethyl acetate in hexane. After reaction completion, the reaction is quenched by slowly adding saturated NaHCO3solution at 0-5°C followed by extraction with n-pentane and washing of organic layer with water and brine. Organic layer is further dried with anhydrous sodium sulphate and concentrated in water bath at a temperature below 15°C to yield compound 2.

[0162] dimethyl 2,3-dihydro-1H-indene-4,5-dicarboxylate (3):

[0163] The crude compound 2 (1-vinylcyclopent-l-ene) is dissolved in benzene, and DMAD (1.2 equivalents) is added to the compound mixture. The reaction mixture is refluxed for 12 hours. Followed by adding DDQ (1.2 equivalents) and benzene at room temperature, and the mixture is again refluxed for another 12 hours. Upon completion of reaction, the reaction is allowed to cool to room temperature and is further concentrated using a rotary evaporator. The resulting product is then purified by column chromatography using 15 to 25% ethyl acetate in hexane as an eluent, to yield compound 3

[0164] 7,8-dihvdrocvcloDentalelisoindole-1,3(2H,6H)-dione (4): P_W0100786

[0165] To a stirred solution of compound 3 (dimethyl 2,3-dihydro-1H-indene-4,5-dicarboxylate), in THF: MeOH: H₂O (3:2:1), NaOH (2.5 equivalents) is added to form a mixture, and the reaction mixture is stirred at room temperature for 5 hours. Then the mixture is concentrated, followed by adding water and is then acidified with concentrated HC1. The mixture slurry is then filtered out and dried, resulting in crude product.

[0166] Urea is added (1.2 equivalents) to the obtained crude product and heated at 150°C for 2 hours to yield a solid compound. The solid formed is then used for further reactions without any purification and the reaction is monitored by TLC using 30% ethyl acetate in hexane, to yield 92% Compound 4

[0167] 5-nitro-7,8-dihydrocyclopentalelisoindole-1,3(2H,6H)-dione (5):

[0168] Astirred solution of nitrating mixture (1:1 H2SO4) is added to the crude compound 4 ((7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) to obtain reaction mixture, and the reaction mixture is continuously stirred for 2 hours with TLC monitoring using 30% ethyl acetate in hexane. Upon completion of reaction, the reaction mixture is slowly added to ice, resulting in a solid compound, then the compound is filtered, dried and purified by column chromatography, yielding 88% Compound 5

[0169] 5-amino-7,8-dihydrocyclopentarelisoindole-1,3(2H,6H)-dione (6):

[0170] Pd / C (10 mol%) is added to a stirred solution of compound 5 (5-nitro-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) in DCM, with continuous stirring under a hydrogen atmosphere at room temperature for 4 hours. Upon completion of reaction, the mixture is filtered out through Celite and further purified by column chromatography, yielding 98% Compound 6

[0171] General procedure for the preparation Compounds 7-41 as mentioned in Examples: Example 1

[0172] 5-((4-methoxybenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (7):

[0173] .0

[0174] HN-4*

[0175] O

[0176]

[0177] ,o

[0178] DMF is added to compound 6 (5-amino-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) in a round-bottomed flask equipped with a charged magnetic stirrer, in argon P_W0100786

[0179] atmosphere to obtain a solution. l-(bromomethyl)-4-methoxybenzene and potassium iodide is added to this solution with continuous stirring at 90 °C for 12 hours to remove solid product, followed by adding Ice water to the solution to filter off the solid product. The resulting product is then crystallized by adding mixture of dichloromethane, methanol and hexane yielding in compound 7

[0180] 87%;1H NMR (400 MHz, DMSO-d6) 5 10.58 (s, 1H), 8.50 (d, J = 5.9 Hz, 2H), 7.33 (d, J = 5.9 Hz, 2H), 6.96 (t, J = 6.3 Hz, 1H), 6.45 (s, 1H), 4.54 (d, J = 5.9 Hz, 2H), 3.04 (t, J = 7.6 Hz, 2H), 2.79 (t, J = 7.4 Hz, 2H), 2.15 (p, J = 7.5 Hz, 2H).

[0181] Example 2

[0182] 5-((4-(trifluoromethyl)phenethyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (8):

[0183] Compound 8 is synthesized by following same procedure as compound 7.

[0184]

[0185] 55%;1H NMR (400 MHz, DMSO-d6) 5 10.58 (s, 1H), 7.65 (d, J = 8.0 Hz, 2H), 7.49 (d, J = 8.0 Hz, 2H), 6.74 (s, 1H), 6.22 (t, J = 5.8 Hz, 1H), 3.57 - 3.47 (m, 2H), 3.03 - 2.93 (m, 4H), 2.63 (t, J = 7.3 Hz, 2H), 2.08 (dd, J = 14.9, 7.6 Hz, 2H).

[0186] Example 3

[0187] 5-((3,5-bis(trifluoromethyl)benzyl) amino)-7,8-dihydrocyclopenta[e]isoindole 1,3 (2H,6H) -dione (9):

[0188] Compound 9 is synthesized by following the same procedure as compound 7.

[0189]

[0190] P_W0100786

[0191] 78%; ’H NMR (400 MHz, DMSO-d6) 5 10.60 (s, 1H), 8.10 (d, J = 6.0 Hz, 2H), 8.00 (s, 1H), 7.00 - 6.92 (m, 1H), 6.64 (s, 1H), 4.68 (d, J =10.1 Hz, 2H), 3.04 (t, J = 7.6 Hz, 2H), 2.79 (t, J = 7.4 Hz, 2H), 2.19 - 2.09 (m, 2H).

[0192] Example 4

[0193] 5-((4-nitrobenzyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (10): Compound 10 is synthesized by following same procedure as compound 7

[0194] HN

[0195] HN

[0196]

[0197] 82%;1H NMR (400 MHz, DMSO-d6) 5 10.58 (s, 1H), 8.21 (d, J = 8.8 Hz, 2H), 7.60 (t, J = 9.2 Hz, 2H), 7.07 - 6.98 (m, 1H), 6.47 (s, 1H), 4.65 (d, J = 6.2 Hz, 2H), 3.03 (t, J = 7.6 Hz, 2H), 2.79 (t, J = 7.4 Hz, 2H), 2.15 (p, J = 7.6 Hz, 2H).

[0198] Example 5

[0199] 5-((2,4,6-trimethylbenzyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (11):

[0200] Compound 11 is synthesized by following same procedure as compound 7.

[0201]

[0202] 85%;1H NMR (400 MHz, DMSO) 5 10.58 (s, 1H), 6.86 (d, 7= 8.4 Hz, 3H), 5.80 (s, 1H), 4.30 (t, J = 9.1 Hz, 2H), 3.01 (t, J = 7.5 Hz, 2H), 2.66 (t, J = 7.4 Hz, 2H), 2.29 (s, 6H), 2.23 (s, 3H), 2.11 -2.01 (m, 2H).

[0203] Example 6 P_W0100786

[0204] 5-((3,5-dinitrobenzyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (12):

[0205] Compound 12 is synthesized by following the same procedure as compound 7

[0206]

[0207] 82%;1H NMR (500 MHz, DMSO-d6) δ 10.60 (s, 1H), 8.71 (t, 7= 2.1 Hz, 1H), 8.69 (d, 7 = 2.1 Hz, 2H), 7.07 (t, 7 = 6.5 Hz, 1H), 6.68 (s, 1H), 4.78 (d, 7 = 6.4 Hz, 2H), 3.04 (t, 7 = 7.6 Hz, 2H), 2.81 (t, 7= 7.5 Hz, 2H), 2.16 (p, 7 = 7.7 Hz, 2H).

[0208] Example 7

[0209] 5-((3-fluoro-4-nitrobenzyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (13):

[0210] Compound 13 is synthesized by following the same procedure as compound 7.

[0211]

[0212] NO2

[0213] 65%; ’H NMR (500 MHz, DMSO-d6) 5 10.59 (s, 1H), 8.15 (t, 7 = 8.1 Hz, 1H), 7.54 (d, 7 = 12.3 Hz, 1H), 7.41 (dd, 7= 8.5, 1.1 Hz, 1H), 6.99 (t, 7= 6.3 Hz, 1H), 6.50 (s, 1H), 4.64 (d, 7 = 6.2 Hz, 2H), 3.08 - 2.99 (m, 2H), 2.80 (t, 7 = 7.4 Hz, 2H), 2.20 - 2.08 (m, 2H). P_W0100786

[0214] Example 8

[0215] 5-((4-(trifluoromethyl) benzyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (14):

[0216] Compound 14 is synthesized by following the same procedure as compound 7.

[0217]

[0218] 72%;1H NMR (400 MHz, DMSO-d6) 5 10.57 (s, 1H), 7.46 (d, J = 8.7 Hz, 2H), 7.33 (d, J = 7.9 Hz, 2H), 7.00 - 6.88 (m, 1H), 6.52 (s, 1H), 4.53 (d, J = 6.2 Hz, 2H), 3.02 (t, J = 7.6 Hz, 2H), 2.77 (t, J = 7.4 Hz, 2H), 2.21 - 2.05 (m, 2H).

[0219] Example 9

[0220] 5-((pyridin-4-ylmethyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (15):

[0221] Compound 15 is synthesized by following the same procedure as compound 7.

[0222]

[0223] 64%; 1H NMR (400 MHz, DMSO-d6) 5 10.58 (s, 1H), 8.50 (d, J = 5.9 Hz, 2H), 7.33 (d, J = 5.9 Hz, 2H), 6.96 (t, J = 6.3 Hz, 1H), 6.45 (s, 1H), 4.54 (d, J = 5.9 Hz, 2H), 3.04 (t, J = 7.6 Hz, 2H), 2.79 (t, J = 7.4 Hz, 2H), 2.15 (p, J = 7.5 Hz, 2H). P_W0100786

[0224] Example 10

[0225] 5-((4-(methylthio) benzyl) amino)-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (16):

[0226] Compound 16 is synthesized by following the same procedure as compound 7.

[0227]

[0228] 78%; 1H NMR (500 MHz, DMSO-d6) 5 10.54 (s, 1H), 7.29 (d, J = 8.1 Hz, 2H), 7.22 (d, J = 8.3 Hz, 2H), 6.90 (t, J = 5.9 Hz, 1H), 6.49 (s, 1H), 4.45 (d, J = 5.6 Hz, 2H), 3.02 (t, J = 7.4 Hz, 2H), 2.76 (t, J = 7.2 Hz, 2H), 2.43 (s, 3H), 2.14 (q, J = 7.3 Hz, 2H).

[0229] ADVANTAGES

[0230] 1. The tricyclic compound of the present invention shows promising efficacy and safe to use as targeted therapy, through selective PARP inhibition in Breast and Ovarian Cancer treatment.

[0231] 2. The tricyclic compound shows enhanced solubility and bioavailability while minimizing the haematological toxicity.

[0232] 3. The synthesized compound is therapeutically useful due to its high tumour selectivity and less side effects.

Claims

P_W0100786We Claim,1. A tricyclic compound of formula I as (Poly (ADP-ribose) Polymerase) PARP inhibitor.RGeneral Formula (I)wherein,n is 0,1, 2, 3, 4, 5 and 6.R is selected from the group consisting of hydrogen, homo or hetero cycloalkyl, optionally substituted with halogen group comprising of-CN, -NO2, -NH2, -SH, -SMe, -OH, OMe, -CF3, -Me. or 3 to 6 membered homo or hetero cycloalkyl, or 5 to 6 membered mono- or bicyclic homo or hetero aromatic rings.wherein, the hetero aromatic ring comprising of heteroatoms is selected from the group consisting of nitrogen, oxygen, Sulfur optionally substituted with halogen group comprising of, CN, NO2, NH2, SH. SMe, OH. OMe. CF3, Me.

2. The compound as claimed in claim 1, wherein the compound is selected from the group consisting of:a) 5-((4-methoxybenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (7)b) 5-((4-(trifluoromethyl)phenethyl)amino)-7,8- dihydrocyclopenta[e]isoindolel,3(2H,6H)-dione (8)c) 5-((3,5-bis(trifluoromethyl)benzyl)amino)-7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (9)d) 5-((4-nitrobenzyl)amino) -7,8 -dihydrocy clopenta[e] isoindole- l,3(2H,6H)-dione (10)e) 5-((2,4,6-trimethylbenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (11)P_W0100786f) 5-((3,5-dinitrobenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (12)g) 5-((3-fluoro-4-nitrobenzyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (13)h) 5 - ((4- (trifluoromethy l)benzy 1) amino)-7, 8 - dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione (14)i) 5-((pyridin-4-ylmethyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (15)j) 5-((4-(methylthio)benzyl)amino)-7,8-dihydrocyclopenta[e]isoindole- l,3(2H,6H)-dione (16)3. A process for preparing the tricyclic compound as claimed in claim 1, wherein the process comprising the steps of:i. adding (1.2 equivalents) Vinyl magnesium bromide to the solution of cyclopentane (compound 1) and dietyl ether at a temperature in the range of 0- 5°C for a time period in the range of 1-1.5 hours, and quenching the reaction using NH4Cl solution followed by extracting with diethyl ether;ii. washing and concentrating the extracted reaction mixture of step (i) to obtain a crude product and followed by dissolving the obtained crude product in n- pentane and p-Toulene sulfonic acid P-TSA, (1.2 equivalents) under nitrogenic conditions at a temperature in the range of 0-5°C for a time period of 1-1.5 hrs; iii. quenching the reaction of step (ii) using saturated NaHCO3solution and extracting with n-pentane and followed by ashing and drying the organic layer with anhydrous sodium sulphate, and further concentrating in water bath at a temperature in the range of 0-15 °C to obtain compound 2 ( 1 -vinylcyclopent- 1- ene).iv. adding Dimethyl acetylenedicarboxylate (DMAD 1.2 equivalents) to the solution of compound 2 as obtained in step (iii) dissolved in benzene and mixing it for a time period in the range of 10-12 hours to obtain a reaction mixture; v. adding 2,3-Dichloro-5,6-dicyano-l,4-benzoquinone (DDQ 1.2 equivalents) and benzene to the reaction mixture obtained in step (iv) at room temperature and mixing it fora time period in the range of 12-14 hrs. Cooling the reaction mixture and purifying using column chromatography (15 to 25% ethyl acetate in hexaneP_W0100786as an eluent), to obtain compound 3 (dimethyl 2,3-dihydro-1H-indene-4,5- dicarboxylate).vi. adding NaOH (2.5 equivalents) to the solution of compound 3 (dimethyl 2,3- dihydro-1H-indene-4,5-dicarboxylate) asobtained in step (v) dissolved in THF: MeOH: H₂O (3:2:1) to form a mixture and stirring the mixture at room temperature for a time period of 5 to 7 hours, and concentrating the reaction mixture;vii. adding water and concentrated HC1 to the mixture of step (vi) to form a slurry and filtering, drying the mixture slurry to obtain crude product followed by adding urea (1.2 equivalents) to the obtained crude product and heating at a temperature in the range of 150°C to 170 °C for a time period of 2 to 4 hours to obtain compound 4 (7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione). viii. adding a solution of nitrating mixture (1:1 H2SO4) to the crude compound 4 ((7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) obtained in step (vii) to form a reaction mixture and stirring it for a time period of 2 to 4 Hours with TLC monitoring using 30% ethyl acetate in hexane;ix. adding the reaction mixture as obtained in step (viii) slowly to ice, resulting in a solid compound. Then filtering, drying and purifying the solid compound using column chromatography to obtain compound 5 (5-nitro-7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione).x. adding Pd / C (10 mol%) to the solution of compound 5 (5-nitro-7,8- dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione) as obtained in step (ix) dissolved in DCM dichloromethane under a hydrogen atmosphere at room temperature for a time period in the range of 4 to 6 Hours. Then filtering and purifying the reaction mixture using column chromatography to obtain Compound 6 (5-amino-7,8-dihydrocyclopenta[e]isoindole-1,3(2H,6H)-dione). xi. adding DMF Dimethylformamide to the compound 6 as obtained in step (x) under argon atmosphere to make a solution. Then adding l-(bromomethyl)-4- methoxybenzene and potassium iodide to the obtained solution and mixing with continuous stirring at a temperature in the range of 90 °C to 120 °C for a time period of 12 to 16 hours to remove solid product;xii. filtering the solution obtained in step (xi) by adding ice water and crystallizing the filtered solution by adding a mixture of dichloromethane, methanol and hexane to obtain the compounds 7-41.P_W01007864. A pharmaceutical composition for treating breast and ovarian cancer comprising of:c) a compound as claimed in claim 2;d) a pharmaceutically acceptable salt thereof.

5. A method of treating breast cancer by administering the pharmaceutical composition as claimed in claim 4, at a particular dosage.

6. Use of the compound as claimed in claim 1-4 for treating breast and ovarian cancer.

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