Methods for treating breast cancer and PARP-resistant breast cancer

Abstract

A method for treating breast cancer comprises a combination of a therapeutically effective amount of irsidin or an irsidin analog, derivative, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a PARP inhibitor. The composition and kit are included herein. Breast cancer can be refractory to various PARP inhibitors.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 368,510, filed on July 15, 2022, the entire content of which is incorporated herein by reference.

[0002] This application relates to the treatment of breast cancer. More particularly, this application relates to the combination treatment of breast cancer in subjects that are refractory or have relapsed, and that have previously been treated with a PARP inhibitor. Further, the combination treatment can be used to treat triple - negative breast cancer.

Background Art

[0003] Breast cancer is the most common cancer in women worldwide and the second leading cause of cancer - related death in women in the United States. Triple - negative breast cancer (TNBC) is a subtype of breast cancer characterized by the absence of estrogen receptor (ER), progesterone receptor, and human epidermal growth factor receptor 2 (HER2). HER2 + In other subtypes of breast cancer, such as tumors, HER2 receptor and ER using drugs that block estrogen signaling + Although significant progress has been made in the development of targeted agents for breast cancer, there is no approved or effective targeted treatment for TNBC. Historically, cytotoxic chemotherapy has been the only viable systemic treatment option for patients with TNBC.

[0004] Triple - negative breast cancer accounts for about 10 - 20% of all breast cancer cases. It tends to occur more frequently in younger women and women with certain genetic mutations such as BRCA1. TNBC often exists as a more aggressive form of breast cancer with a higher likelihood of recurrence and metastasis (spread to other organs) compared to other breast cancer subtypes. Due to the absence of hormone receptors and HER2 overexpression, treatment options for TNBC are limited to chemotherapy, surgery, and radiation therapy.

[0005] Therefore, there is always a need for improved treatments for breast cancer. In particular, this need is the subject of the present application. SUMMARY OF THE INVENTION

[0006] The present application discloses the discovery of a treatment for breast cancer in a subject using a combination of irinotecan or an irinotecan analog (e.g., asulfaben) and a PARP inhibitor. Such treatment may have a greater effect than the effect provided by either asulfaben or PARP inhibitor treatment alone.

[0007] One aspect of the present application includes a combination therapy for treating cancer. In embodiments, the therapy includes administering a combination of active agents including an irinotecan or an irinotecan analog (e.g., asulfaben) and a PARP inhibitor. The method includes administering to a subject in need of treatment a combination of active agents having a therapeutically effective amount of irinotecan or an irinotecan analog, derivative, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a PARP inhibitor or a pharmaceutically acceptable salt thereof. The subject may have recurrent cancer and / or refractory cancer. The subject may have been previously treated with a PARP inhibitor. The subject may be treated in combination after the cancer has become refractory and resistant to the PARP inhibitor.

[0008] One aspect includes a method where the subject has relapsed after about 1 month, 2 months, 3 months, 4 months, or more after discontinuation of treatment with a PARP inhibitor.

[0009] Another aspect includes a method where the irinotecan analog is asulfaben.

[0010] Another aspect includes a method where the irinotecan analog is hydroxyurea methyl asulfaben.

[0011] Another aspect includes a method where the irinotecan analog has the following structure.

[0012]

Chem.

[0013] Another aspect includes a method in which the irudin analog has the following structure.

[0014]

Chem.

[0015] Another aspect includes a method in which the irudin analog is irofulven.

[0016] Another aspect of the present application provides a pharmaceutical composition comprising an irudin or an irudin analog (e.g., acylfulven) and a PARP inhibitor or a pharmaceutically acceptable salt thereof, mixed with a pharmaceutically suitable carrier or excipient, in a dose for treating or preventing cancer, mainly in a person who has been previously treated with a PARP inhibitor. The pharmaceutical composition can also be administered simultaneously, sequentially, or alternately in combination with other therapeutic agents or treatment modalities.

[0017] Another aspect of the present application includes a therapy comprising acylfulven which is (-)-hydroxyurea methyl acylfulven.

Brief Description of the Drawings

[0018]

Figure 1

Figure 2

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Figure 6

BRIEF DESCRIPTION OF THE INVENTION

[0019] The present application provides a method of treating breast cancer in a subject. One embodiment includes a method of treating a person having breast cancer and breast cancer resistant to a PARP inhibitor, the method comprising administering an effective amount of an acylfulvene (e.g., hydroxyureamethylacylfulvene) or a salt thereof and a PARP inhibitor.

[0020] Iludin or acylfulvene In one embodiment, the present application includes the use of iludin or an iludin analog (e.g., acylfulvene). Acylfulvene (also known as 6-acetylfulvene) is a synthetic anticancer drug belonging to the class of alkylating agents. It is derived from a natural product called fulvoic acid, which is found in plants and soil. Acylfulvene is a cytotoxic semi-synthetic derivative of iludin, a natural product that can be extracted from the jack-o'-lantern mushroom (Omphalotus olearius). Acylfulvene, derived from sesquiterpene iludin S by treatment with acid (the retro-Prins reaction), is much less reactive towards thiols than iludin S. Acylfulvene acts by alkylating DNA, which means that acylfulvene forms a covalent bond with DNA molecules inside cells, causing damage and interfering with replication and transcription processes. This action prevents cancer cells from dividing and ultimately induces cell death.

[0021] In one embodiment, the acylfulvene is (-)-hydroxyurea methyl acylfulvene (referred to as LP-184 by Lantern Pharma Inc.), which positively shifts the light as shown below.

[0022]

Chemical formula

[0023] In another embodiment, the acylfulvene is (+)-hydroxyurea methyl acylfulvene (referred to as LP-284 of Lantern Pharma Inc.), which positively shifts the light as shown below.

[0024]

Chemical formula

[0025] (+)-Hydroxyurea methyl acylfulvene and (-)-hydroxyurea methyl acylfulvene are enantiomers and are currently publicly known.

[0026] In another example, the acylfulvene is ilofulvene.

[0027] LP-284 and LP-184 are enantiomers and are currently publicly known.

[0028] In another example, the acylfulvene is ilofulvene or 6-hydroxymethyl acylfulvene.

[0029]

Chemical formula

[0030] PARP inhibitor PARP inhibitors are a type of cancer drug. PARP stands for poly-adenosine diphosphate-ribose polymerase (poly-ADP ribose polymerase (PARP) inhibitor), and is a type of enzyme that helps repair DNA damage in cells. PARP inhibitors act by preventing cancer cells from repairing damaged DNA and causing cancer cells to die. The PARP enzyme helps repair DNA damage. By blocking this, cancer cells can be prevented from being repaired, thereby killing cancer cells. There are at least four major PARP inhibitors: olaparib (Lynparza), niraparib (Zejula), rucaparib (Rubraca), and talazoparib (Talzenna). Pharmaceutical compositions are disclosed that include one or more PARP inhibitors and typically at least one additional substance, such as an excipient, a known therapeutic agent other than those of the present disclosure, and combinations thereof. In some embodiments, the PARP inhibitor can be used in combination with other agents known to have beneficial, additive, or synergistic activity with the PARP inhibitor. In one particular embodiment, the PARP inhibitor is a PARP-1 inhibitor. In other embodiments, the PARP inhibitor is an inhibitor of any enzyme of the PARP family, such as PARP1 and / or PARP2.

[0031] Examples of suitable PARP inhibitors according to the present invention include, but are not limited to, olaparib (AZD-2281, 4-[(3-[(4-cyclopropylcarbonyl)piperazin-4-yl]carbonyl)-4-fluorophenyl]methyl(2H)-phthalazin-1-one), veliparib (ABT-888, CAS 912444-00-9, 2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carboxamide), CEP-8983 (11-methoxy-4,5,6,7-tetrahydro-1H-cyclopenta[a]pyrrolo[3,4-c]carbazole-1,3(2H)-dione) or its prodrug (e.g., CEP-9722), rucaparib (AG014699, PF-01367338, 8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[-5,4,3-cd]indol-6-one), E7016 (GPI-21016, 10-((4-hydroxypiperidin-1-yl)methyl)chromeno-[4,3,2-de]phthalazin-3(2H)-one), talazoparib (BMN-673, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,-9-dihydro-2H-pyrido[4,3,2de]phthalazin-3(7H)-one), INO-1001 (4-phenoxy-3-pyrrolidin-1-yl-5-sulfamoyl-benzoic acid), KU0058684 (CAS 623578-11-0), niraparib (MK4827, Merck & Co Inc), iniparib (BSI201), iniparib-met (C-nitroso metabolite of iniparib), CEP9722 (Cephalon Inc), LT-673, MP-124, NMS-P118, XAV939, AZD2461, nicotinamide, 5-methylnicotinamide, 4-amino-1,8-naphthalimide, picolinamide, benzamide, 3-substituted benzamide, 3-methoxybenzamide, 3-hydroxybenzamide, 3-aminobenzamide, 3-chloroprocaine amide, 3-nitrosobenzamide, 4-aminobenzamide, 2-aminobenzamide, methyl 3,5-diiodo-4-(4'-methoxyphenoxy)benzoate, methyl-3,5-Iodo-4-(4'-methoxy-3',5'-diiodo-phenoxy)benzoate, cyclic benzamide, 1,5-di[(3-carbamoylphenyl)aminocarbonyloxy]pentane, indole, benzimidazole, benzoxazole-4-carboxamide, benzimidazole-4-carboxamide, 2-substituted benzoxazole 4-carboxamide, 2-substituted benzimidazole 4-carboxamide, 2-arylbenzimidazole 4-carboxamide, 2-cycloalkylbenzimidazole-4-carboxamide, 2-(4-hydroxyphenyl)benzimidazole A-carboxamide, quinoxaline carboxamide, imidazopyridine carboxamide, 2-phenylindole, 2-substituted benzoxazole, 2-phenylbenzoxazole, 2-(3-methoxyphenyl)benzoxazole, 2-substituted benzimidazole, 2-phenylbenzimidazole, 2-(3-methoxyphenyl)benzimidazole, 1,3,4,5-tetrahydro-azepino[5,4,3-cd]indol-6-one, azepinoindole, azepinoindrone, 1,5-dihydro-azepino[4,5,6-cd]indolin-6-one, dihydrodiazepinoindolinone, 3-substituted dihydrodiazepinoindolinone, 3-(4-trifluoromethylphenyl)-dihydrodiazepinoindolinone, tetrahydrodiazepinoindolinone, 5,6-dihydroimidazo[4,5,1-j,k][1,4]benzodiazepin-7(4H)-one, 2-phenyl-5,6-dihydro-imidazo[4,5,1-jk][1,4]benzodiazepin-7(4H)-one, 2,3-dihydro-isoindol-1-one, benzimidazole-2-piperazine, benzimidazole-2-piperazine heterocyclic derivative, 4-iodo-3-nitrobenzamide, benzopyrone, 1,2-benzopyrone 6-nitroso-benzopyrone, 6-nitroso 1,2-Benzopyrone, 5-iodo-6-aminobenzopyrone, benzoylurea, quinolone, isoquinolone, isoquinolinone, dihydroisoquinolinone, 2H-isoquinolin-1-one, 3H-quinazolin-4-one, 5-substituted dihydroisoquinolinone, 5-hydroxy dihydroisoquinolinone, 5-methyldihydroisoquinolinone, 5-hydroxyisoquinolinone, 5-aminoisoquinolin-1-one, 5-dihydroxyisoquinolinone, 1,5-dihydroxyisoquinoline, 1,5-isoquinolinediol, 4-hydroxyquinazoline, substituted thiazolyl-isoquinolinone, substituted oxazolyl-isoquinolinone, tetrahydro-2H-isoquinolin-1-one, 3,4-dihydroisoquinolin-1(2H)-one, 3,4-dihydro-5-methoxy-isoquinolin-1(2H)-one, 3,4-dihydro-5-methyl-1(2H)isoquinolinone, 3H-quinazolin-4-one, isoquinolin-1(2H)-one, 3,4-dihydroisoquinolin-1(2H)-one, 4-carboxamido-benzimidazole, substituted 6-cyclohexylalkyl substituted 2-quinolinone, substituted 6-cyclohexylalkyl substituted 2-quinoxalinone, 7-phenylalkyl substituted 2-quinolinone, 7-phenylalkyl substituted 2-quinoxalinone, 6-substituted 2-quinolinone, 6-substituted 2-quinoxalinone, 1-(arylmethyl)quinazoline-2,4(1H,3H)-dione, 4,5-dihydro-imidazo[4,5,1-ij]quinolin-6-one, 1,6-naphthyridin-5(6H)-one, 1,8-naphthalimide, 4-amino-1,8-naphthalimide, 3,4-dihydro-5-[4-1(1-piperidinyl)butoxy]-1(2H)-isoquinolinone, 2,3-dihydrobenz[de]isoquinolin-1-one, 1-1 Ib-dihydro-[2H]benzopyrano[4,3,2-de]isoquinolin-3-one, tetracyclic lactam, benzopyranoisoquinolinone, benzopyrano[4,3,2-de]isoquinolinone, quinazoline, quinazolinone, quinazoline dione, A-hydroxyquinazoline, 2-substituted quinazoline, 8-hydroxy-2-methylquinazolin-4-(3H)one, phthalazine, phthalazinone, phthalazin-1(2H)-one, 5-methoxy-4-methyl-1(2)phthalazinone, 4-substituted phthalazinone, 4-(1-piperazinyl)-1(2H)-phthalazinone, tetracyclic benzopyrano[4,3,2-de]phthalazinone and tetracyclic indeno[1,2,3-de]phthalazinone, tricyclic phthalazinone, 2-aminophthalhydrazide, phthalazinone ketone, dihydropyridophthalazinone, 6-substituted 5-aryl amino-1h-pyridin-2-one, pyridazinone, tetrahydropyridopyridazinone, tetraazaphenalen-3-one, thieno[2,3-c]isoquinolin-5-one (TIQ-A), 2,5-diazabicyclo[2.2.1]heptane, pyrimidoimidazole, isoindolinone, phenanthridine, phenanthridinone, 5[H]phenanthridin-6-one, substituted 5[H]phenanthridin-6-one, 2,3-substituted 5[H]phenanthridin-6-one, sulfonamide / carbamide derivatives of 6(5H)phenanthridinone, thieno[2,3-c]isoquinolone, 9-aminothieno[2,3-c]isoquinolone, 9-hydroxy thieno[2,3-c]isoquinolone, 9-methoxy thieno[2,3-c]isoquinolone, N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-2-(N,N-dimethylamino)acetamide, substituted 4,9-dihydrocyclopenta[imn]phenanthridin-5-one, unsaturated hydroxamic acid derivatives, O-(3-piperidino-2-hydroxy-1-propyl)nicotinic acid amidoxime, O-(2-hydroxy-3-piperidino-propyl)-3-carboxylic acid amidoxime, pyridazine, pyrazinamid, BGB-290, PF-1367338 (Pfizer Inc), AG014699 (Pfizer, Inc.), KU-59436 (KuDOS / AstraZeneca PJ34, 4-amino-1,8-naphthalimide (Trevigen), 6(5H)-phenanthridinone (Trevigen), NU1025, 4-HQN, BGP-15, A-966492, GPI21016, 6(5H)-phenanthridinone (Phen), theobromine, theophylline, caffeine, methylxanthine, thymidine, 3-aminophthalhydrazide, analogs, derivatives or mixtures thereof are included.,

[0032] Additional PARP inhibitors include, for example, WO 2019 / 14201972, WO 2019 / 14201972, WO 2019 / 12141990, WO 2019 / 10091140, WO 2019 / 09524379, WO 2019 / 09155402, WO 2019 / 009046205, WO 2019 / 08146035, WO 2019 / 08015429, WO 2019 / 0191796, WO 2019 / 0042040, US 2006 / 004028, EP 2604610, EP 1802578, CN 104140426, CN 104003979, US 2006 / 0229351, US 7,041,675, WO 2019 / 07041357, WO 2019 / 03057699, US 2006 / 0444676, US 2006 / 229289, US 2006 / 0063926, WO 2019 / 06033006, WO 2019 / 06033007, WO 2019 / 03051879, WO 2019 / 04108723, WO 2019 / 06066172, WO 2019 / 06078503, US 2007 / 0032489, WO 2019 / 05023246, WO 2019 / 05097750, WO 2019 / 05123687, WO 2019 / 05097750, US 7,087,637, US 6,903,101, WO 2019 / 070011962, US 2007 / 0015814, WO 2019 / 06135873, UA 2007 / 0072912, WO 2019 / 06065392, WO 2019 / 05012305, WO 2019 / 05012305, EP 412848, EP 453210, EP 454831, EP 879820, EP 879820, WO 2019 / 030805, WO 2019 / 03007959, US 6,989,388, US 2006 / 0094746, EP 1212328, WO 2019 / 06078711, US 2006 / 0426415, US 2006 / 0514,No. 983, European Patent No. 1212328, US Patent Application Publication No. 20040254372, US Patent Application Publication No. 20050148575, US Patent Application Publication No. 20060003987, US Patent Application No. 06 / 635,642, International Publication No. 200116137, International Publication No. 2004105700, International Publication No. 03057145 (A2), International Publication No. 2006078711, International Publication No. 2002044157, US Patent Application Publication No. 20056924284, International Publication No. 2005112935, US Patent Application Publication No. 20046828319, International Publication No. 2005054201, International Publication No. 2005054209, International Publication No. 2005054210, International Publication No. 2005058843, International Publication No. 2006003146, International Publication No. 2006003147, International Publication No. 2006003148, International Publication No. 2006003150, International Publication No. 2006003146, International Publication No. 2006003147, UA20070072842, US Patent Application No. 05 / 587,384, US Patent Application Publication No. 20060094743, International Publication No. 2002094790, International Publication No. 2004048339, European Patent No. 1582520, US Patent Application Publication No. 20060004028, International Publication No. 2005108400, US Patent No. 6,964,960, International Publication No. 20050080096, International Publication No. 2006137510, UA20070072841, International Publication No. 2004087713, International Publication No. 2006046035, International Publication No. 2006008119, International Publication No. 06008118, International Publication No. 2006042638, US Patent Application Publication No. 20060229289, US Patent Application Publication No. 20060229351, International Publication No. 2005023800, International Publication No. 1991007404, International Publication No. 2000042025, International Publication No. 2004096779, US Patent No. 6,426,415, International Publication No. 2068407, US Patent No. 6,476,No. 048, International Publication No. WO2001090077, International Publication No. WO2001085687, International Publication No. WO2001085686, International Publication No. WO2001079184, International Publication No. WO2001057038, International Publication No. WO2001023390, International Publication No. WO01021615(A1), International Publication No. WO2001016136, International Publication No. WO2001012199, International Publication No. WO95024379, International Publication No. WO200236576, International Publication No. WO2004080976, International Publication No. WO2007149451, International Publication No. WO2006110816, International Publication No. WO2007113596, International Publication No. WO2007138351, International Publication No. WO2007144652, International Publication No. WO2007144639, International Publication No. WO2007138351, International Publication No. WO2007144637, Banasik et al. (J. Biol. Chem., 267:3, 1569-75, 1992), Banasik et al. (Molec. Cell. Biochem, 138:185-97, 1994), Cosi et al. (Expert Opin. Ther. Patents 12(7), 2002), Southan and Szabo (Curr Med Chem, 10 321-340, 2003), Underhill C. et al. (Annals of Oncology, doi:10.1093 / annonc / mdq322, pp 1-12, 2010), Murai J. et al. (J. Pharmacol. Exp. Ther., 349:408-416, 2014), all of these patents and publications are hereby incorporated by reference in their entirety.,

[0033] In a preferred embodiment, the PARP inhibitor compound is selected from the group consisting of rucaparib (AG014699, PF-01367338), olaparib (AZD2281), veliparib (ABT888), iniparib (BSI201), niraparib (MK4827), talazoparib (BMN673), AZD2461, CEP9722, E7016, INO-1001, LT-673, MP-124, NMS-P118, XAV939, their analogs, derivatives, or mixtures.,

[0034] In an even more preferred embodiment, the PARP inhibitor is selected from the group consisting of rucaparib, olaparib, veliparib, iniparib, niraparib, talazoparib, AZD2461, their analogs, derivatives, or mixtures.

[0035] In one embodiment, the acylfulvene or hydroxyurea methyl acylfulvene or a salt thereof may be administered either before, simultaneously with, or after the administration of the PARP inhibitor.

[0036] One aspect of the present application includes a method of doing so in a subject in need of treating cancer. The method involves administering to the subject an effective amount of a PARP inhibitor and an effective amount of an acylfulvene. The PARP inhibitor can be administered before or simultaneously with the acylfulvene for optimal synergistic effect.

[0037] Another embodiment includes a pharmaceutical composition having a therapeutically effective amount of irudin or an irudin analog, derivative, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a PARP inhibitor or an analog, derivative, or pharmaceutically acceptable salt thereof. The irudin analog can be hydroxyurea methyl acylfulvene.

[0038] In another embodiment, a kit for treating cancer in a subject includes a therapeutically effective amount of irudin or an irudin analog, derivative, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a PARP inhibitor or an analog, derivative, or pharmaceutically acceptable salt thereof.

[0039] In one embodiment, breast cancer can be treated with the following regimen: histologically confirmed breast cancer, PARP-resistant / non-responsive disease can be defined as disease progression (resistance) within 180 days after the last administered dose of PARP therapy, or lack of response or disease progression (non-responsiveness) during recent PARP-based therapy, respectively.

[0040] PARP inhibition causes synthetic lethality in germline BRCA and BRCA - related breast cancers and is routinely used in the clinical treatment of metastatic breast cancer. It may also be beneficial for breast cancers with homologous recombination deficiency or BRCAness, most commonly triple - negative breast cancer. Currently, the use of PARP inhibitors for triple - negative breast cancer with wild - type BRCA does not have conclusive efficacy. One embodiment includes treating breast cancer by administering an effective amount of acylfulvene or hydroxyurea methyl acylfulvene or a salt thereof.

[0041] In one example, the tumor cells are NERD or NER tumors. NER - deficient cancers refer to a group of cancers associated with defects in the nucleotide excision repair (NER) pathway. If there are defects or mutations in the NER pathway, it can lead to the accumulation of DNA damage, which may increase the risk of developing certain cancers. NER is a cellular mechanism involved in the repair of DNA damage caused by environmental factors such as UV irradiation and certain chemicals. NERD or NER - deficient tumors are nucleotide excision repair - deficient tumors, and their phenotypes are the result of mutations in genes involved in DNA repair, including but not limited to ERCC1, ERCC3, ERCC4, ERCC5, ERCC6, RAD50, ATR, ATM, MRE, CSB, XPD, etc.

[0042] In one embodiment, the patient is treated with radiation prior to treatment with acylfulvene.

[0043] In another embodiment, the second therapeutic agent can be one or more chemotherapeutic agents selected from camptothecin derivatives, paclitaxel, docetaxel, epothilone B, 5 - FU, gemcitabine, oxaliplatin, cisplatinum, carboplatin, melphalan, dacarbazine, temozolomide, doxorubicin, imatinib, erlotinib, bevacizumab, cetuximab, and Raf kinase inhibitors.

[0044] In another embodiment, the second therapeutic agent is one or more chemotherapeutic agents selected from paclitaxel or cisplatinum.

[0045] The treatment can be the "first choice", i.e., the first treatment in patients who have not previously received anti-cancer treatment, either alone or in combination with other treatments; or it can be the "second choice" as a treatment in patients who have received one previous anti-cancer treatment regimen, either alone or in combination with other treatments; or it can be the "third choice", "fourth choice", etc., as a treatment performed either alone or in combination with other treatments.

[0046] In another embodiment, a kit for treating cancer in a subject comprises a therapeutically effective amount of irinotecan or an irinotecan analog, derivative, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a PARP inhibitor or an analog, derivative, or pharmaceutically acceptable salt thereof.

[0047] In another embodiment, the second therapeutic agent is one or more chemotherapeutic agents selected from camptothecin derivatives, paclitaxel, docetaxel, epothilone B, 5-FU, gemcitabine, oxaliplatin, cisplatinum, carboplatin, melphalan, dacarbazine, temozolomide, doxorubicin, imatinib, erlotinib, bevacizumab, cetuximab, and Raf kinase inhibitors.

[0048] In another embodiment, the second therapeutic agent is one or more chemotherapeutic agents selected from paclitaxel or cisplatinum.

[0049] The term "combination therapy" can include, or can include the administration of the therapeutic agents described above in further combination with other biologically active ingredients and non-drug therapies (e.g., surgery or radiation therapy). Where combination therapy further includes non-drug therapy, the non-drug therapy can be carried out at any appropriate time as long as a beneficial effect from the concurrent action of the combination of the therapeutic agent and the non-drug therapy is achieved. For example, in appropriate cases, a beneficial effect can be achieved even if the non-drug therapy is temporarily (possibly for several days or even weeks or months) removed from the administration of the therapeutic agent.

[0050] In another aspect, the compositions or combination therapies herein, or pharmaceutically acceptable salts or solvates thereof, may be administered in combination with radiation therapy. Radiation therapy may also be administered in combination with the compositions of the invention and other chemotherapeutic agents described herein as part of a multi-drug therapy.

[0051] Combination therapy can be achieved by administering two or more agents, e.g., acylfulvene, a PARP inhibitor, and one or more other therapeutic agents, each of which can be formulated and administered separately, or can be achieved by administering two or more agents in a single formulation. Other combinations are also encompassed by combination therapy. For example, two agents can be formulated together and administered in combination with a separate formulation containing a third agent. The two or more agents in combination therapy can be administered simultaneously, but they need not be. For example, administration of the first agent (or combination of agents) can precede administration of the second agent (or combination of agents) by minutes, hours, days, or weeks. Thus, the two or more agents can be administered within minutes of each other, or within 1, 2, 3, 6, 9, 12, 15, 18, or 24 hours of each other, or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14 days of each other, or within 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks of each other. In some cases, longer intervals are possible. In many cases, it is desirable for two or more agents used in combination therapy to be present in the patient's body simultaneously, but this is not necessary.

[0052] A method of combination therapy should or may result in a synergistic effect where the effect of the combination of a compound or other therapeutic agent is greater than the sum of the effects resulting from the administration of either the compound or the other therapeutic agent as a single agent. The synergistic effect may also be an effect that cannot be achieved by administering either the compound or the other therapeutic agent as a single agent. Examples of synergistic effects can include, but are not limited to, the effect of treating cancer by reducing tumor size, inhibiting tumor growth, or increasing the survival of a subject. The synergistic effect may also include reducing cancer cell survival, inducing cancer cell death, and inhibiting or delaying cancer cell growth.

[0053] The therapeutically effective dose can vary depending on the disease being treated, the severity of the disease, the route of administration, the age and general health of the patient, the use of excipients, the possibility of combination with other therapeutic treatments such as the use of other drugs, and the judgment of the treating physician, as recognized by those skilled in the art. For example, guidance for selecting an effective dose can be determined by referring to the prescribing information of acylfulvene or hydroxyurea methyl acylfulvene or the discussion in its journal.

[0054] As used herein, the term "effective amount" refers to the amount of an agent necessary to reduce at least one or more symptoms of a disease or disorder, and relates to the amount of a pharmacological composition sufficient to achieve the desired effect. Thus, the term "therapeutically effective amount" refers to the amount of an agent that, when administered to a typical subject, is sufficient to achieve a particular effect. An effective amount can be an amount sufficient to reduce the symptoms of a disease responsive to inhibition of a PARP inhibitor. In the case of cancer treatment, in vivo efficacy can be measured, for example, by evaluating survival, time to disease progression (TTP), response rate (RR), duration of response, and / or quality of life. An effective amount can vary depending on the route of administration, use of excipients, and co-administration with other agents, as will be appreciated by those skilled in the art. An effective amount in various circumstances also includes, as used herein, an amount sufficient to delay the onset of symptoms of a disease, modify the course of disease symptoms (e.g., without limitation, slow the progression of disease symptoms), or reverse the symptoms of a disease. Thus, it is generally not practical to specify an exact "effective amount". However, in any given case, an appropriate "effective amount" can be determined by one of ordinary skill in the art using routine experimentation only.

[0055] The dosage range for administration of an agent in accordance with the methods described herein depends, for example, on the form of the agent, its potency, and the degree to which it is desirable to reduce symptoms, markers or indicators of the conditions described herein, e.g., the desired rate of decrease in tumor growth. The dosage should not be so large as to cause harmful side effects. Generally, the dosage can vary depending on the age, condition, and sex of the patient and can be determined by one of ordinary skill in the art. The dosage can also be adjusted by the individual physician in the event of any complications.

[0056] For example, the efficacy of the agents described herein in the treatment of the conditions described herein, or for inducing the responses (e.g., solid or hematological cancers) described herein, can be determined by a skilled clinician. However, if one or more of the signs or symptoms of the conditions described herein change beneficially, other clinically acceptable symptoms improve or even resolve, or the desired response is induced by at least 10% for example, after treatment by the methods described herein, the treatment is considered, when this term is used herein, an “effective treatment”. For example, efficacy can be evaluated by measuring markers, indicators, symptoms and / or incidence rates of the condition being treated by the methods described herein, or any other measurable appropriate parameter, such as tumor size and / or growth rate. Efficacy can also be measured by the adverse event of an individual deteriorating upon hospitalization, or the need for medical intervention (i.e., the progression of the disease is halted). Methods for measuring these indicators are known to those of skill in the art and / or are described herein. Treatment includes any treatment of a disease in an individual or animal (some non-limiting examples include humans or animals), (1) inhibiting the disease, e.g., preventing the worsening of symptoms (e.g., pain or inflammation), or (2) reducing the severity of the disease, e.g., causing regression of symptoms. An effective amount for the treatment of a disease means an amount sufficient, when administered to a subject in need thereof, to provide an effective treatment as defined herein for that disease. The effectiveness of an agent can be determined by evaluating physical indicators of the condition or desired response. Monitoring the efficacy of administration and / or treatment by measuring any one of such parameters, or any combination of parameters, is well within the ability of those of skill in the art. Efficacy can be evaluated in animal models of the conditions described herein, for example, in the treatment of hematological cancers in a mouse model. When using experimental animal models, the efficacy of treatment is evidenced by observing a statistically significant change in a marker, such as tumor size and / or tumor growth rate.In some embodiments, the therapeutically effective amount of hydroxyurea methyl-acylfulvene, acylfulvene, ilofulvene, or a pharmaceutically acceptable salt thereof is selected from the group consisting of 0.5 mg / day, 1 mg / day, 2.5 mg / day, 5 mg / day, 10 mg / day, 20 mg / day, 30 mg / day, 60 mg / day, 90 mg / day, 120 mg / day, 150 mg / day, 180 mg / day, 210 mg / day, 240 mg / day, 270 mg / day, 300 mg / day, 360 mg / day, 400 mg / day, 440 mg / day, 480 mg / day, 520 mg / day, 580 mg / day, 600 mg / day, 620 mg / day, 640 mg / day, 680 mg / day, and 720 mg / day.

[0057] The dosage should be adjusted according to the requirements of the individual in need. The dosage of PARP (poly ADP-ribose polymerase) inhibitors can vary depending on the specific drug used and the medical condition for which it is prescribed. PARP inhibitors are a type of targeted therapy used in the treatment of certain types of cancer, particularly cancers with defects in the DNA repair mechanism. For illustration, olaparib (Lynparza): In the case of ovarian cancer, a typical dosage is 300 mg twice daily (total daily dosage 600 mg). For breast cancer, for those with germline BRCA mutations, it is 300 mg twice daily (total daily dosage 600 mg). Further, niraparib (Zejula): A typical dosage for maintenance therapy of ovarian cancer is 300 mg once daily. The dosage can be adjusted based on individual patient factors. Further, rucaparib (Rubraca): A typical dosage for ovarian cancer and other related indications is 600 mg twice daily (total daily dosage 1200 mg). The actual dosage prescribed to a patient can be determined by a healthcare provider based on various factors including the specific cancer being treated, the overall health of the patient, and other individual considerations.

[0058] The term "treat" is used to include both therapeutic treatment and prophylactic treatment (reducing the likelihood of development). Both terms mean reducing, suppressing, attenuating, eliminating, arresting, or stabilizing the development or progression of a disease (e.g., a disease or disorder described herein), reducing the severity of a disease, or ameliorating the symptoms associated with a disease.

[0059] The pharmaceutical composition can be contained in a container, pack, or dispenser together with instructions for administration.

[0060] The compositions of the present invention can further form salts. The compositions of the present invention can form more than one salt per molecule, such as mono-salts, di-salts, tri-salts. All of these forms are also considered to be within the scope of the claimed invention.

[0061] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the compounds of the present invention, where the parent compound is modified by making its acid or base salts. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues (e.g., amines), and alkali or organic salts of acidic residues (e.g., carboxylic acids). Pharmaceutically acceptable salts include, for example, conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic and organic acids selected from, but not limited to, 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonate, citric acid, edetic acid, ethanedisulfonic acid, 1,2-ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, glycolsulfonic acid, hexylresorcinolic acid, hydrabamic acid, hydrobromic acid, etc.

[0062] Other examples of pharmaceutically acceptable salts include hexanoic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, muconic acid, and the like. The present invention also encompasses salts formed when an acidic proton in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth ion or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like.

[0063] All references to pharmaceutically acceptable salts are to be understood to include solvent addition forms (solvates) of the same salts.

[0064] As used herein, the term "selectively" means having a tendency to occur at a higher frequency in one population than in another. The populations being compared can be cell populations. Preferably, if an event occurs at a frequency more than 2-fold higher in population A compared to population B, it occurs selectively in population A relative to population B. An event occurs selectively if it occurs at a frequency more than 5-fold higher in population A. An event occurs more preferably selectively if it occurs at a frequency more than 10-fold higher in population A compared to population B, even more preferably if it occurs at a frequency more than 50-fold higher, still more preferably if it occurs at a frequency more than 100-fold higher, and most preferably if it occurs at a frequency more than 1000-fold higher. For example, cell death is said to occur selectively in cancer cells if it occurs more frequently in cancer cells than in normal cells by more than 2-fold.

[0065] The composition, or a pharmaceutically acceptable salt or solvate thereof, is administered orally, nasally, transdermally, by the pulmonary, inhalation, buccal, sublingual, intraperitoneal, subcutaneous, intramuscular, intravenous, rectal, intrapleural, intrathecal, and parenteral routes. In one embodiment, the compound is administered orally. One of ordinary skill in the art will recognize the advantages of a particular route of administration.

[0066] The dosage regimen utilizing the compounds is selected according to a variety of factors including the type, species, age, weight, sex and medical condition of the patient; the severity of the condition being treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof being employed. A ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, reverse, or arrest the progress of the condition.

[0067] Techniques for formulation and administration of the disclosed compounds of the invention can be found in Remington: the Science and Practice of Pharmacy, 19 th edition, Mack Publishing Co., Easton, Pa. (1995). In one embodiment, the compounds and pharmaceutically acceptable salts thereof described herein are used in pharmaceutical preparations in combination with a pharmaceutically acceptable carrier or diluent. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous or organic solutions. The compounds are present in such pharmaceutical compositions in an amount sufficient to provide the desired dosage within the ranges described herein.

[0068] All percentages and ratios used herein are by weight unless otherwise indicated. Other features and advantages of the invention will be apparent from the different examples. The examples provided illustrate different components and methodologies useful in practicing the invention. The examples do not limit the invention as claimed. Based on the present disclosure, one of ordinary skill in the art can identify and use other components and methodologies useful in practicing the invention.

[0069] As used herein, a "subject in need thereof" is a subject having a pre-cancerous condition. Preferably, the subject in need thereof has cancer. The "subject" includes mammals. The mammal can be, for example, any mammal, such as a human, primate, bird, mouse, rat, dog, cat, cow, horse, goat, camel, sheep or pig. Preferably, the mammal is a human. The subjects of the present invention include any human subject diagnosed with cancer or a pre-cancerous condition, having symptoms of cancer or a pre-cancerous condition, or at risk of developing cancer or a pre-cancerous condition.

[0070] The subject in need thereof can have refractory cancer or resistant cancer. "Refractory cancer or resistant cancer" means cancer that does not respond to treatment. The cancer may be resistant at the start of treatment or may become resistant during treatment. In some embodiments, the subject in need thereof has cancer recurrence after remission with a recent therapy. In some embodiments, the subject in need thereof has received and failed all known effective therapies for cancer treatment. In some embodiments, the subject in need thereof has received at least one previous therapy. In certain embodiments, the previous therapy is a monotherapy. In certain embodiments, the previous therapy is a combination therapy.

[0071] "Recurrent cancer" is cancer that has been treated before and as a result of that treatment, the subject had completely or partially recovered (i.e., the subject is said to be in a remission state), but the cancer has recurred or progressed after the treatment was discontinued. In other words, recurrent cancer is cancer that was responsive to treatment and became resistant to treatment after a period during which the subject had completely or partially recovered. Typically, in patients with breast cancer, the development of resistant and refractory disease occurs after multiple rounds of treatment. Additionally, PARP inhibitors can become refractory and lead to treatment resistance when cancer cells develop mutations that bypass the mechanism of action of the drug or alter alternative signaling pathways. This can occur at various stages of treatment and can vary depending on the type of cancer being treated and the individual response of the patient to the drug.

[0072] In some embodiments, the subject in need thereof may have secondary cancer as a result of previous therapy. "Secondary cancer" means cancer that results from or is caused by a previous carcinogenic therapy such as chemotherapy.

[0073] Cancer is a group of diseases that can cause almost any sign or symptom. The signs and symptoms depend on where the cancer is, the size of the cancer, and how much it affects nearby organs or structures. If the cancer spreads (metastasizes), symptoms can appear in different parts of the body.

[0074] Treatment of cancer can reduce the size of the tumor. A reduction in tumor size may also be referred to as "tumor regression". Preferably, after treatment, the tumor size is reduced by at least 5% compared to its size before treatment, more preferably, the tumor size is reduced by at least 10%, more preferably, by at least 20%, more preferably, by at least 30%, more preferably, by at least 40%, even more preferably, by at least 50%, and most preferably, by at least 75%. The tumor size may be measured by any reproducible measurement means. The tumor size may be measured as the diameter of the tumor.

[0075] Treatment of cancer reduces the number and size of tumors. Preferably, after treatment, the number or size of tumors is reduced by at least 5% compared to the number before treatment, more preferably, the number or size of tumors is reduced by at least 10%, more preferably, by at least 20%, more preferably, by at least 30%, more preferably, by at least 40%, even more preferably, by at least 50%, and most preferably, by more than 75%. The number of tumors may be measured by any reproducible measurement means. The number of tumors may be measured by counting the tumors that are visible to the naked eye or at a specific magnification. Preferably, the specific magnification is 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or 50-fold.

[0076] Treatment of cancer can result in a decrease in the number of metastatic lesions in other tissues or organs distant from the primary tumor site. Preferably, after treatment, the number of metastatic lesions is reduced by at least 5% compared to the number before treatment, more preferably by at least 10%, more preferably by at least 20%, more preferably by at least 30%, more preferably by at least 40%, even more preferably by at least 50%, and most preferably by more than 75%. The number of metastatic lesions may be measured by any reproducible means of measurement. The number of metastatic lesions may be measured by counting the metastatic lesions, which are visible to the naked eye or at a specific magnification. Preferably, the specific magnification is 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or 50-fold.

[0077] Treatment of cancer can increase the average survival time of a population of treated subjects compared to a population receiving the carrier alone. Preferably, the average survival time is increased by more than 30 days, more preferably by more than 60 days, more preferably by more than 90 days, and most preferably by more than 120 days. The increase in the average survival time of the population may be measured by any reproducible means. The increase in the average survival time of the population can be measured, for example, by calculating the average length of survival for the population after the start of treatment with the active compound. The increase in the average survival time of the population can also be measured, for example, by calculating the average length of survival for the population after the completion of the first round of treatment with the active compound.

[0078] Treatment of cancer can result in an increase in the average survival period of the treated population compared to the untreated population. Preferably, the average survival period increases by more than 30 days, more preferably by more than 60 days, more preferably by more than 90 days, and most preferably by more than 120 days. The increase in the average survival period of the population may be measured by any reproducible means. The increase in the average survival period of the population can be measured, for example, by calculating the average length of survival for the population after the start of treatment with the active compound. The increase in the average survival period of the population can also be measured, for example, by calculating the average length of survival for the population after the completion of the first round of treatment with the active compound.

[0079] Treatment of cancer can result in an increase in the average survival period of the treated population compared to a population receiving monotherapy with a compound of the invention or a drug that is not a pharmaceutically acceptable salt or solvate thereof. Preferably, the average survival period increases by more than 30 days, more preferably by more than 60 days, more preferably by more than 90 days, and most preferably by more than 120 days. The increase in the average survival period of the population may be measured by any reproducible means. The increase in the average survival period of the population can be measured, for example, by calculating the average length of survival for the population after the start of treatment with the active compound. The increase in the average survival period of the population can also be measured, for example, by calculating the average length of survival for the population after the completion of the first round of treatment with the active compound.

[0080] Treating cancer can result in a decrease in the lethality rate of the population to be treated compared to a population receiving the carrier alone. Treating cancer can result in a decrease in the lethality rate of the population to be treated compared to an untreated population. Treating cancer can result in a decrease in the lethality rate of the population to be treated compared to a population receiving monotherapy with a compound of the present invention or a drug that is not a pharmaceutically acceptable salt or solvate thereof. Preferably, the lethality rate is decreased by more than 2%, more preferably by more than 5%, more preferably by more than 10%, and most preferably by more than 25%. The decrease in the lethality rate of the population to be treated may be measured by any reproducible means. The decrease in the lethality rate of the population can be measured, for example, by calculating the average number of disease-related deaths per unit time for the population after the start of treatment with the active compound. The decrease in the lethality rate of the population can also be measured, for example, by calculating the average number of disease-related deaths per unit time for the population after the completion of the first round of treatment with the active compound.

[0081] Treating cancer can result in a decrease in the tumor growth rate. Preferably, after treatment, the tumor growth rate is decreased by at least 5% relative to the pre-treatment value, more preferably the tumor growth rate is decreased by at least 10%, more preferably by at least 20%, more preferably by at least 30%, more preferably by at least 40%, more preferably by at least 50%, even more preferably by at least 50%, and most preferably by at least 75%. The tumor growth rate may be measured by any reproducible measurement means. The tumor growth rate can be measured by the change in the diameter of the tumor per unit time.

[0082] Treatment of cancer can result in a reduction in tumor regrowth. Preferably, after treatment, tumor regrowth is less than 5%, more preferably less than 10%, more preferably less than 20%, more preferably less than 30%, more preferably less than 40%, more preferably less than 50%, even more preferably less than 50%, and most preferably less than 75%. Tumor regrowth may be measured by any reproducible measurement means. Tumor regrowth is measured, for example, by measuring the increase in the diameter of the tumor after previous tumor shrinkage following treatment. A reduction in tumor regrowth is indicated by the tumor not recurring after treatment is stopped.

[0083] Treatment or prevention of a cell proliferative disorder can result in a reduction in the rate of cell proliferation. Preferably, after treatment, the cell proliferation rate is reduced by at least 5%, more preferably at least 10%, more preferably at least 20%, more preferably at least 30%, more preferably at least 40%, more preferably at least 50%, even more preferably at least 50%, and most preferably at least 75%. The rate of cell proliferation may be measured by any reproducible measurement means. The rate of cell proliferation is measured, for example, by measuring the number of dividing cells in a tissue sample per unit time.

[0084] Treatment or prevention of a cell proliferative disorder can result in a reduction in the proportion of proliferating cells. Preferably, after treatment, the proportion of proliferating cells is reduced by at least 5%, more preferably at least 10%, more preferably at least 20%, more preferably at least 30%, more preferably at least 40%, more preferably at least 50%, even more preferably at least 50%, and most preferably at least 75%. The proportion of proliferating cells may be measured by any reproducible measurement means. Preferably, the proportion of proliferating cells is measured, for example, by quantifying the number of dividing cells relative to the number of non-dividing cells in a tissue sample. The proportion of proliferating cells may be equivalent to the mitotic index.

[0085] The treatment or prevention of a cell proliferative disorder can result in a decrease in the size of the region or zone of cell proliferation. Preferably, after treatment, the size of the region or zone of cell proliferation is reduced by at least 5% relative to its size before treatment, more preferably at least 10% reduction, more preferably at least 20% reduction, more preferably at least 30% reduction, more preferably at least 40% reduction, more preferably at least 50% reduction, even more preferably at least 50% reduction, most preferably at least 75% reduction. The size of the region or zone of cell proliferation may be measured by any reproducible measuring means. The size of the region or zone of cell proliferation may be measured as the diameter or width of the region or zone of cell proliferation.

[0086] The treatment or prevention of a cell proliferative disorder can result in a decrease in the number or proportion of cells having an abnormal appearance or geometry. Preferably, after treatment, the number of cells having an abnormal morphology is reduced by at least 5% relative to its size before treatment, more preferably at least 10% reduction, more preferably at least 20% reduction, more preferably at least 30% reduction, more preferably at least 40% reduction, more preferably at least 50% reduction, even more preferably at least 50% reduction, most preferably at least 75% reduction. The appearance or morphology of abnormal cells may be measured by any reproducible measuring means. The geometry of abnormal cells can be measured, for example, by using an inverted tissue culture microscope. The geometry of abnormal cells can take the form of nuclear pleomorphism.

[0087] By treating cancer or a cell proliferative disorder, cell death can be induced, preferably, the cell death results in at least a 10% decrease in the number of cells in the population. More preferably, the cell death means at least a 20% decrease, more preferably at least a 30% decrease, more preferably at least a 40% decrease, more preferably at least a 50% decrease, most preferably at least a 75% decrease. The number of cells in the population may be measured by any reproducible means. The number of cells in the population can be measured by fluorescence activated cell sorting (FACS), immunofluorescence microscopy and light microscopy. The method for measuring cell death is as shown in Li et al., Proc. Natl. Acad. Sci. USA. 100(5):2674-8, 2003. In one embodiment, the cell death is caused by apoptosis.

[0088] Preferably, an effective amount of the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, is not significantly cytotoxic to normal cells. A therapeutically effective amount of a compound is not significantly cytotoxic to normal cells if administration of the compound at a therapeutically effective amount does not induce more than 10% cell death of normal cells. A therapeutically effective amount of a compound does not significantly affect the survival rate of normal cells if administration of the compound at a therapeutically effective amount does not induce more than 10% cell death of normal cells. In one embodiment, the cell death is caused by apoptosis.

[0089] By contacting cells with the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, selective induction or activation of cell death can be achieved in cancer cells. By administering the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, to a subject in need thereof, selective induction or activation of cell death can be achieved in cancer cells. By contacting cells with the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, selective induction of cell death can be achieved in one or more cells affected by a cell proliferative disorder. Preferably, by administering the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, to a subject in need thereof, selective induction of cell death is achieved in one or more cells affected by a cell proliferative disorder.

[0090] The present invention relates to a method for treating or preventing cancer by administering the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, to a subject in need thereof, wherein the administration of the composition of the present invention, or a pharmaceutically acceptable salt or solvate thereof, results in prevention of cancer cell proliferation by accumulation of cells in one or more phases of the cell cycle (e.g., G1, G1 / S, G2 / M), or induction of cellular senescence, or promotion of tumor cell differentiation, promotion of cell death in cancer cells by cytotoxicity, necrosis or apoptosis without significant amounts of cell death in normal cells, and having anti-tumor activity with a therapeutic index of at least 2. As used herein, "therapeutic index" is the ratio of the maximum tolerated dose to the effective dose.

[0091] The term "kit" means that the combination partners defined above can be administered independently or by use of different fixed combinations having an identified amount of combination partner, i.e., simultaneously or at different times. Then, the parts of a kit of parts can be administered, for example, simultaneously or shifted over time, i.e., at different times for any of the parts of the kit of parts, at equal or different time intervals. The ratio of the total amounts of combination partners administered in a combination preparation can vary. The combination partners can be administered by the same route or different routes.

[0092] One of ordinary skill in the art can refer to general reference texts for a detailed description of the known or equivalent techniques discussed herein. These texts can, of course, also be referred to when making or using aspects of the present invention.

[0093] LP-184 and LP-284 belong to the family of acylfulvene compounds known to induce DNA damage that is repaired by the Transcription-Coupled Nucleotide Excision Repair (TC-NER) pathway.

Examples

[0094] The following non-limiting examples illustrate the methods of the present disclosure.

[0095] Example 1 Triple-negative breast cancer (TNBC), a subtype of breast cancer that does not express estrogen receptor (ER), progesterone receptor (PR), or human epidermal growth factor receptor 2 (HER-2), is characterized by high invasiveness, high metastatic potential, high recurrence rate, and poor prognosis. Due to its special molecular phenotype, TNBC is not sensitive to endocrine therapy or targeted therapy. Therefore, chemotherapy is the main systemic treatment, but the effectiveness of conventional postoperative adjuvant chemoradiotherapy is poor. LP-184, an acylfulvene-derived prodrug, is activated by the oxidoreductase PTGR1 and is synthetically lethal in tumors with DNA damage repair defects including homologous recombination (HR) deficiency for the treatment of solid tumor indications including TNBC.

[0096] Cell viability assays were used to generate LP-184 IC50 in breast cancer cell lines in vitro. A subcutaneous patient-derived TNBC xenograft mouse model was used to determine the tumor volume response in vivo. The xenograft tumors were derived from treatment-naive HR-deficient primary TNBC patients with known BRCA1 / 2 LOH that had progressed on the PARP inhibitor olaparib.

[0097] Figure 1 shows that nanomolar concentrations of LP-184 inhibited the in vitro viability of six NCI-60 breast cancer cell lines (median IC50 = 327 nM).

[0098] Example 2 - LP-184 Activity in Combination with a PARP Inhibitor in Triple-Negative Breast Cancer PDX models of TNBC have been established by several groups using various methods and usually involve engrafting tumor fragments or dissociated tumor cells from frozen or fresh core needle biopsies into the mammary glands of immunodeficient mice. Such models are widely used to evaluate the preclinical assessment of any new therapeutic approach. The PDX model for this study was a TNBC tumor (invasive ductal carcinoma) transplanted into the flanks of female NOD scid gamma (NSG) mice. Tumor size was measured every 7 days. Treatment with LP-184 or hydroxyurea methylacylfulvene resulted in complete tumor regression in a PDX model of TNBC that was HR-deficient and resistant to PARP inhibitors and doxorubicin / cyclophosphamide.

[0099] Patient-derived xenograft tumors of triple-negative breast cancer were transplanted subcutaneously into mice. Two PDX models of the TNBC model, the PARPi-sensitive model HBCx10 (BRCA2 variant) and the PARPi-resistant model HBCx28 (BRCA1 variant), were treated with different dose levels of LP-184 and / or the PARP inhibitor olaparib. 60 - 200 mm 3Mice with established growing tumors were randomly assigned to each treatment arm. Tumor volume and mouse body weight were measured twice a week. For HBCx10, LP-184 was administered intravenously on days 1 and 8, and olaparib was administered orally daily for 21 days. For HBCx28, LP-184 was administered intravenously on days 1, 4, 8, and 11, and olaparib was administered orally daily for 21 days. In HBCx10, LP-184 treatment at 2 mg / kg (days 1 and 8 of a 21-day cycle) was equivalent to daily olaparib at 80 mg / kg and had a similar trend of tumor regression in each case. The combination of LP-184 (0.75 mg / kg) + olaparib (40 mg / kg) showed the highest synergy compared to the individual treatments. Similarly, the combination of LP-184 (0.75 mg / kg) + olaparib (80 mg / kg) was synergistic. These results are shown in Figures 2 and 3, which measure HBC×10 tumor volume vs. time in TNBC during treatment with LP-184 and / or olaparib. Figure 3 shows the HBCx10 tumor volume in the PDX HBCx10 model for treatment with LP-184 and / or olaparib in HRD-positive PARPi-sensitive tumors. Figure 4 shows the tumor volume in the PDX HBCx10 model over treatment with LP-184 and olaparib.

[0100] On day 22, the following tumor growth inhibition rate (TGI %) values shown in Table 1 below demonstrate the synergy between LP-184 (0.75 mg / kg) + olaparib (40 mg / kg).

[0101]

Table 1

[0102] In HBCx28, LP-184 treatment at 2 mg / kg (days 1, 4, 8, 11 or twice weekly over 2 weeks) was more potent than any olaparib dose level. The combination of LP-184 (0.75 mg / kg) + olaparib (80 mg / kg) was synergistic.

[0103] Furthermore, on the 21st day, the tumor growth inhibition rate (TGI %) values shown in Table 2 below demonstrate the synergistic effect between LP-184 (0.75 mg / kg) and olaparib (80 mg / kg). LP-184 alone at 2 and 4 mg / kg shows the highest tumor regression.

[0104]

Table 2

[0105] Example 4 - Synergistic effect Figures 4 and 5 show that LP-184 and olaparib exhibit a synergistic effect in TNBC HBCx10 and HBCx28 models, respectively. The tumor volume in HBCx10 with LP-184 treatment at 2 mg / kg (on the 1st and 8th days of a 21-day cycle) was equivalent to that with daily olaparib at 80 mg / kg, and in each case, there was a similar trend of tumor regression. The combination of LP-184 (0.75 mg / kg) + olaparib (40 mg / kg) showed the highest synergistic effect compared to individual treatments.

[0106] Example 4 - PARP inhibitor resistance In the absence of any valid genetically engineered mouse model for TNBC, the PDX model is widely used to evaluate the preclinical assessment of any new therapeutic approach. Figure 6 shows complete tumor regression in HR-deficient, multi-drug resistant (PARP inhibitor resistant) TNBC in the PDX HBCx-24 model. The PDX model for this study was one in which a TNBC tumor (invasive ductal carcinoma) was transplanted into the flanks of female NOD scid gamma (NSG) mice. Tumor size was measured every 7 days.

[0107] Complete tumor regression was observed in the HR-deficient multi-drug resistant TNBC PDX model HBCx-24 with 4 mg / kg LP-184 i.v. treatment over 2 cycles, and the 107% TGI was on the 32nd day (Figure 6).

Claims

1. A method for treating breast cancer that is PARP - refractory or resistant in a patient in need of treatment, said method comprising administering a combination to a subject in need of treatment, said combination comprising a. a therapeutically effective amount of irudine or an irudine analog, derivative, or a pharmaceutically acceptable salt thereof, and b. a therapeutically effective amount of a PARP inhibitor or a pharmaceutically acceptable salt thereof, wherein the subject has recurrent cancer and / or refractory cancer, the subject has been previously treated with the PARP inhibitor, and the subject is treated with the combination after the cancer has become refractory and resistant to the PARP inhibitor.

2. The method according to claim 1, wherein the subject relapsed more than about 1 month after discontinuation of treatment with the PARP inhibitor.

3. The method according to claim 1, wherein the irudine analog is acylfulvene.

4. The irudine analog is hydroxyurea methyl acylfulvene, the method according to claim 1.

5. The method according to claim 1, wherein the irudine analog has the following structure. 【Chemical 1】

6. The method according to claim 1, wherein the irudine analog has the following structure. 【Chemical 2】

7. The method according to claim 1, wherein the irudine analog is ilofulvene.

8. The method according to claim 1, wherein the PARP inhibitor is administered after at least two rounds of treatment with the PARP inhibitor.

9. The method according to claim 1, wherein the PARP inhibitor or a pharmaceutically acceptable salt, solvate or hydrate thereof, and the irudine or irudine analog are for separate, simultaneous, or sequential use or administration.

10. The method according to claim 1, wherein the PARP inhibitor is administered at a dose of 200 - 1200 mg per day.

11. The method according to claim 1, wherein the irudine analog and the PARP inhibitor are administered separately.

12. The method according to claim 1, wherein the irudine analog and the PARP inhibitor are administered daily.

13. The method according to claim 1, wherein the irudine analog and the PARP inhibitor are administered continuously.

14. The method according to claim 1, wherein the active agents are administered as a co - formulation.

15. The method according to claim 1, wherein the administration of irudine or an analog thereof is before, during, or after the administration of the PARP inhibitor.

16. The method according to claim 1, further comprising subjecting the subject to radiotherapy, chemotherapy, or performing surgery on the subject before, during, or after administration of the iridium and / or the PARP inhibitor.

17. The method according to claim 1, wherein one or more of the PARP inhibitors are selected from the group consisting of olaparib, rucaparib, and niraparib.

18. The method according to claim 1, wherein the subject is an animal.

19. The method according to claim 1, wherein the subject or mammal is a human.

20. The method according to claim 5, further comprising subjecting the subject to radiotherapy before, after, or during treatment with hydroxyurea methyl acylfulvene.

21. The method according to claim 1, further comprising administering an additional therapeutic agent selected from the group consisting of cisplatin, paclitaxel, and other available therapies.

22. The method according to claim 1, wherein the breast cancer is triple-negative breast cancer.

23. A method of treating breast cancer, the method comprising administering a combination to a subject in need thereof, the combination comprising a. a therapeutically effective amount of iridium or an iridium analog, derivative, or a pharmaceutically acceptable salt thereof, and b. a therapeutically effective amount of a PARP inhibitor or a pharmaceutically acceptable salt thereof, wherein the breast cancer is triple-negative breast cancer.

24. A kit for treating cancer in a subject, comprising a therapeutically effective amount of iridium or an iridium analog, derivative, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a PARP inhibitor or an analog, derivative, or a pharmaceutically acceptable salt thereof, wherein the subject has recurrent cancer and / or refractory cancer, the subject has been previously treated with ibrutinib, and the subject is treated in combination after the cancer has become refractory and resistant to ibrutinib.

25. The pharmaceutical composition according to claim 24, wherein the iridium analog is hydroxyurea methyl acylfulvene.

26. The iridium analog has the following structure: [Chemical Formula 3] The pharmaceutical composition according to claim 24, having the structure.

27. The iridium analog has the following structure 【Chemical 4】 The pharmaceutical composition according to claim 30, having the structure.

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