Cyclopentenone-based anticancer agents

Abstract

The present invention discloses novel 5-substituted 4-hydroxycyclopent-2-en-1-one derivatives as active compounds in pharmaceutical compositions for cancer treatment. The present invention confers the cytotoxic effect of the above compounds on ovarian cancer, colorectal cancer, cervical cancer, hepatocellular carcinoma, lung cancer, bladder cancer, breast cancer, melanoma, lymphoma, leukemia, and myeloma malignant cells for inhibiting cell cycle progression in the G2 / M phase, reducing the expression of DNA replication licensing factors, and having a general cancer treatment effect. The present invention further discloses the use of the above compositions for the treatment of platinum-resistant tumors. Another aspect of the present invention is the synergistic effect of these compounds with existing cancer therapeutic agents as proteasome inhibitors. Finally, a method for synthesizing the active compounds of the above compositions is disclosed.

Description

Technical Field

[0001] The present invention relates to a method for treating cancer by administering a novel cyclopentenone derivative as an active compound in a pharmaceutical composition. The present invention inhibits cell cycle progression in the G2 / M phase, reduces the expression of DNA replication licensing factors, and has a general cancer therapeutic effect, thereby conferring a cytotoxic effect of the above compound on ovarian cancer, colorectal cancer, cervical cancer, hepatocellular carcinoma, lung cancer, bladder cancer, breast cancer, melanoma, lymphoma, and myeloma malignant cells. The present invention also relates to the use of such compounds in the treatment of abnormal cell proliferation in mammals, particularly humans, and to pharmaceutical compositions as anti-cancer agents. The present invention further discloses the use of the above composition for the treatment of platinum-resistant tumors. Another aspect of the present invention is the synergistic effect of these compounds with existing cancer therapeutic agents as proteasome inhibitors. Finally, a method for synthesizing the active compound of the above composition is disclosed.

Background Art

[0002] Cancer is a group of related diseases in which some of the body's cells grow uncontrollably and spread to other parts of the body. The treatment of these diseases includes surgery, radiotherapy, chemotherapy, immunotherapy, and combinations thereof, and is important for extending the survival of patients. Chemotherapeutic agents are chemical molecules that can be used to treat cancer. Examples include, but are not limited to, platinum-based compounds (cisplatin, carboplatin, oxaliplatin), mitomycin C, bleomycin, topotecan, irinotecan, docetaxel, paclitaxel, vincristine, plicamycin, daunorubicin, adriamycin, 5-fluorouracil, and hormone antagonists.

[0003] Chemotherapeutic agents can be selected based on the type of cancer being treated, the expression of one or more molecular markers by the cancer, the age and overall health status of the patient being treated, etc. For example, high-grade serous epithelial ovarian cancer (EOC) is typically addressed by surgery, followed by adjuvant platinum (cisplatin or carboplatin) and taxane combination chemotherapy. Today, one of the most commonly used, well-known chemotherapeutic drugs is cisplatin. This is because cisplatin treats a wide range of different types of cancers such as carcinomas, germ cell tumors, lymphomas, and sarcomas in different regions of the human body.

[0004] However, due to drug resistance and numerous undesirable side effects (i.e., severe kidney damage, allergic reactions, immunosuppression against infections, gastrointestinal disorders, bleeding, and hearing loss), several other platinum-containing anticancer drugs (e.g., oxaliplatin) are also being used as common cancer treatment active chemotherapeutic agents. These platinum-based drugs are available for a wide range of cancer types but have the same problems as cisplatin such as the development of resistance. As an example, most EOC patients (about 75%) experience disease recurrence within the first 5 years despite aggressive treatment at diagnosis. Topotecan and liposomal doxorubicin are currently one of the second treatment options for platinum-resistant ovarian cancer, with an objective response rate of approximately 10% - 15% in platinum-resistant patients, a median progression-free survival of 9.1 - 13.6 weeks, and an overall survival of only 35.6 - 41.3 weeks [1]. Therefore, resistance to platinum-based chemotherapy is a major clinical problem with limited treatment options. Other therapeutic agents used in EOC include angiogenesis inhibitors and PARP inhibitors that can only be administered to a subset of patients expressing specific markers, but they have been reported to reduce the response to subsequent platinum-based chemotherapy [2].

[0005] Another example is colorectal cancer, a major cause of cancer deaths worldwide. Some of the drugs commonly used to treat colorectal cancer include 5-fluorouracil (5-FU), capecitabine, a tablet that changes to 5-FU when it reaches the tumor, irinotecan, oxaliplatin, and combinations thereof. Sometimes, chemotherapy drugs are administered in combination with targeted therapy drugs. The fixed-dose combination drug Lonsurf is used as a third and fourth treatment for metastatic colorectal cancer after chemotherapy and targeted therapy have failed.

[0006] The clinical application of chemotherapy agents is effective but limited by their systemic or organ-specific toxicity profiles, sub-optimal pharmacokinetic properties, and low stability in vivo. Repurposing of platinum drugs using liposomes has led to the development of L-NDPP (Arroplatin (trademark)), SPI-77, Lipoplatin (trademark), Lipoxal (trademark), and LiPlacIs (registered trademark), which have attractive biological activities such as biocompatibility and improved pharmacokinetics. Pegylated doxorubicin (Doxil (registered trademark) or the European trademark Caelyx (registered trademark)) shows improved in vivo distribution and lower cardiotoxicity.

[0007] Prior art treatment methods have the problem of lacking low cytotoxic activity against cancer cells, specific cytotoxic activity against certain cancer types, for example, resistance after a certain period such as after treatment with cisplatin, and / or the inability to prevent cancers with "translation addiction". The latter term is derived from a model proposed by Elledge and co-workers [3], in which the tumorigenic state depends on gene products that are not the natural cancer genes themselves, but their functions are "rate-limiting" for the survival and proliferation of transformed cells. For example, phosphorylated eIF4E (p-eIF4E) is not required for global translation, but is necessary for transformation and optimal tumorigenesis in tumor models. Graff and co-workers at Eli Lilly Research Labs developed an antisense oligonucleotide against eIF4E, which almost completely blocked tumor growth in breast and prostate xenografts in the absence of a major impact on global protein synthesis [4, 5]. Tumors with overactive MYC increase ribosome biogenesis and are very vulnerable to inhibition of translation [5]. SUMMARY OF THE INVENTION

[0008] The main advantages of the 5-substituted 4-hydroxycyclopent-2-en-1-one derivatives (hereinafter referred to as "enones") of the disclosed compositions, compared to the prior art, are as follows. 1) Broad-spectrum cytotoxic activity due to the high efficacy of enones against different cancer types in vitro and in vivo. 2) The possibility of treating platinum-resistant tumors because cisplatin-resistant ovarian cancer cell lines do not show cross-resistance to 4-hydroxy-4-methyl-5-(naphthalen-2-yl)cyclopent- 2-en-1-one (referred to as enone-1) or 5-(4-fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-en-1-one (referred to as enone-2). 3) Since enone-1 and enone-2 decrease the expression of CDC6 and CDT1 in human tumor cells, there is a possibility of improving the treatment outcome in tumors with increased expression of replication licensing factors such as CDC6 and CDT1. 4) Since enone-1 and enone-2 are inducers of the p53 pathway mediated by the ribosome stress pathway, there is a possibility of improving the treatment outcome in "translation-dependent" tumors. 5) Since enone-2 has a synergistic effect with bortezomib in the death of tumor cells, there is a potential enhancement of proteasome inhibitor therapy.

[0009] The general formula and synthetic route of the novel 4-hydroxycyclopent-2-en-1-one derivatives, which are the active compounds of the claimed composition, have been published [6, 7]. However, 4-hydroxycyclopent-2-en-1-one derivatives having a naphthyl group at the 5-position of the carbon, as well as the specific compounds 5-(4-fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-en-1-one or 4-hydroxy-4-methyl-5-(naphthalen-2-yl)cyclopent-2-en-1-one, which have the highest cytotoxic activity, are difficult to synthesize due to the steric hindrance of the naphthyl group, and thus have not been reported in the literature, nor have their specific synthetic routes been reported.

Mode for Carrying Out the Invention

[0010] The present invention discloses a treatment method using a 5-substituted 4-hydroxycyclopent-2-en-1-one derivative as a drug, more specifically, a treatment method for various types of cancers based on the treatment of a subject with a composition containing an active compound (a compound mainly involved in the cytotoxic effect). More specifically, a method for treating abnormal cell growth, specifically cancer, in a mammal (subject) has the structure:

[0011]

Chemical Formula

[0012] In one embodiment, the method comprises administering a pharmaceutical composition comprising a compound (4 - hydroxycyclopent - 2 - en - 1 - one derivative) having the above structure to a subject, R 1 is phenyl, 4 - fluorophenyl, naphthalen - 2 - yl, 4 - fluoronaphthalen - 1 - yl, CO2Et, CO2Me, n - butyl, R 2 is H, methyl, n - C 16 H 33 is benzyl, n - pentyl, n - hexyl, pent - 4 - enyl, R 3 is H, methyl, R 4 is H, methyl. This is because the cytotoxic effect of the above group of 4 - hydroxycyclopent - 2 - en - 1 - one derivatives is higher, preferably having mainly (more than 50% of the trans isomer) trans stereochemistry, and as a result, in most cases, the 4 - hydroxy group at the 5 - position is trans to the substituent.

[0013] In a preferred embodiment, the method has the structure:

[0014]

Chemical formula

[0015] In another embodiment, the method further comprises administering to a subject a pharmaceutical composition comprising a delivery vehicle that enhances the cellular uptake and / or stability of the compound so as to be suitable and stable for use as a drug. In one embodiment, the delivery vehicle is a liposome, but other delivery vehicles known from the literature are also suitable.

[0016] In another embodiment, the method requires administering the above composition in combination with a proteasome inhibitor, preferably the proteasome inhibitor bortezomib. This is due to the synergistic effect between these compounds and existing cancer therapeutic agents as proteasome inhibitors.

[0017] In another aspect of the present invention, the method comprises administering the above composition to a subject having malignant cell proliferation (cancer), and the above malignant cell proliferation is resistant to platinum-based chemotherapy, preferably cisplatin treatment and / or oxaliplatin, due to its ability to overcome this resistance.

[0018] In a further aspect of the present invention, the method is to administer the above composition to a subject having abnormal cell proliferation, and the above abnormal cell proliferation exhibits translation dependence.

[0019] In a preferred embodiment, the method is used when the cancer is selected from the group consisting of liver cancer, lung cancer, pancreatic cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, breast cancer, cervical cancer, bladder cancer, lymphoma, leukemia, myeloma, or a combination of one or more of the aforementioned cancers.

[0020] In a further embodiment, the inventors claim, for the treatment of abnormal cell growth, preferably cancer, compounds belonging to the above general type having the formula:

[0021] [Chemical formula] preferably the trans isomer (the 4-hydroxy group is trans to the substituent in the 5-position).

[0022] Another aspect of the invention is the administration of the above composition (a composition comprising one or more 5-substituted 4-hydroxycyclopent-2-en-1-one derivatives) (anticancer agent) for the treatment of cancer in a pharmaceutical composition, wherein the pharmaceutical composition comprises the above compound or its tautomer or its pharmaceutically acceptable salt, and a delivery vehicle such as a liposome that enhances the stability of the compound and / or improves the pharmacokinetics and toxicity profile in organs.

[0023] The third aspect of the invention relates to the compounds themselves and their synthesis. 4-Hydroxycyclopent-2-en-1-one derivatives having a naphthyl group at the 5-position of the carbon, specifically the compounds 5-(4-fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-en-1-one or 4-hydroxy-4-methyl-5-(naphthalen-2-yl)cyclopent-2-en-1-one (having the highest cytotoxicity), have not been reported in the literature nor disclosed. Also, due to the steric hindrance of the naphthyl group, synthesis is difficult, and thus their specific synthetic routes are not mentioned. The disclosed synthetic route of the 4-hydroxycyclopent-2-en-1-one derivative was modified. Specifically, the concentration of rose bengal, a photosensitizer, was increased from 0.0001 M to 0.0015 M, and the irradiation time was increased from 3 minutes to 5 minutes. This enabled the synthesis of these novel compounds, which are claimed as drugs.

Brief Description of the Drawings

[0024]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Example

[0025] Synthesis of 4-hydroxy-4-methyl-5-(naphthalen-2-yl)cyclopent-2-en-1-one (2, enone-1) Step 1, Synthesis of 2-methyl-5-(naphthalen-2-ylmethyl)furan (1).

[0026]

Chemical formula

[0027] A solution of 2-methylfuran (670 μL, 7.46 mmol) in anhydrous THF (8 mL) was added dropwise with a solution of n-BuLi (1.6 M in hexane, 4.24 mL, 6.78 mmol) under an argon atmosphere and at 0 °C. The solution was stirred at the same temperature for an additional 30 minutes. Then, a solution of 2-(bromomethyl)naphthalene (750 mg, 3.4 mmol) in anhydrous THF (4 mL) was added dropwise at 0 °C. Subsequently, the solution was warmed to room temperature and stirred for 3 hours. After completion of the reaction as indicated by tlc analysis (3 hours), the reaction was quenched with a saturated aqueous solution of NH4Cl (10 mL), and the resulting mixture was extracted with Et2O (20 mL). The organic layer was separated, dried over Na2SO4, and concentrated in vacuo. The product was purified by flash column chromatography (silica gel, petroleum ether) to obtain 1 as a pale yellow oil (yield = 560 mg, 74%).

[0028] Characteristic NMR spectrum 11H NMR (500 MHz, CDCl3): δ 7.81 (m, 3H), 7.70 (s, 1H), 7.46 (m, 2H), 7.40 (dd, J1 = 8.4 Hz, J2 = 2.0 Hz, 1H), 5.92 (d, J = 2.6 Hz, 1H), 5.90 (m, 1H), 4.10 (s, 2H), 2.27 (s, 3H) ppm; 13 13C NMR (125 MHz, CDCl3): δ 152.6, 151.1, 135.9, 133.6, 132.2, 128.0, 127.6 (2C), 127.3, 127.0, 125.9, 125.4, 107.0, 106.0, 34.7, 13.5 ppm.

[0029] Synthesis of 2 - 4 - hydroxy - 4 - methyl - 5 - (naphthalen - 2 - yl) cyclopent - 2 - en - 1 - one (2, enone - 1)

[0030]

Chemical Structure

[0031] Furan 1 (111 mg, 0.5 mmol) was dissolved in MeOH (5 mL, 0.1 M, in a test tube) containing a catalytic amount (7.3 mg, 1.5% mol) of rose bengal as a photosensitizer. The solution was cooled in an ice bath. While irradiating the solution with a xenon Variac Eimac Cermax 300 W lamp, oxygen was gently bubbled through the solution. After completion of the reaction (5 minutes, indicated by tlc), the solution was warmed to room temperature and Me2S (146 μL, 2 mmol) was added. After completion of the reduction (40 minutes, indicated by tlc analysis), Et3N (21 μL, 0.15 mmol) was added and the reaction mixture was stirred at room temperature for 30 minutes. The reaction was observed by tlc analysis and 1 1H - NMR. After completion of the reaction (30 minutes), the solvent was removed in vacuo and the residue was purified by flash column chromatography (silica gel, petroleum ether:EtOAc = 3:1 → 2:1) to give 2 as a white solid (yield = 103 mg, 87%). To scale up the process, the 5 - minute irradiation step was repeated 10 times on the above scale, the photo - oxidized solutions were combined, and subsequently treated with Me2S and Et3N.

[0032] Characteristic NMR spectrum 1 H NMR (500 MHz, CDCl3): 7.80 (m, 3H), 7.60 (s, 1H), 7.47 (m, 3H), 7.16 (dd, J1 = 8.4 Hz, J2 = 1.5 Hz, 1H), 6.26 (d, J = 5.8 Hz, 1H), 3.94 (s, 1H), 1.03 (s, 3H) ppm; 13 C NMR (125 MHz, CDCl3): 206.3, 165.9, 133.3, 132.7, 132.5, 132.1, 129.0, 128.1, 127.8, 127.6, 127.4, 126.1, 125.9, 80.4, 65.5, 25.7 ppm.

[0033] HRMS (Orbitrap ESI): [M + H] + C 16 H 15 Calculated value for C

[0034] Typical NOE of Compound 2

[0035]

Chemical Structure

[0036] Synthesis of 5-(4-Fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-en-1-one (7, Enone-2) Step 1: Synthesis of Methyl 4-Fluoro-1-naphthoate (3)

[0037]

Chemical Structure

[0038] 4-Fluoro-1-naphthoic acid (580 mg, 3.05 mmol) was dissolved in MeOH (20 mL, 0.15 M) containing sulfuric acid (160 μL, 3 mmol). The solution was heated to reflux in an oil bath overnight until the starting compound was completely consumed. The mixture was quenched with a saturated aqueous solution of NH4Cl (15 mL), and the resulting mixture was extracted with Et2O (30 mL). The organic layer was separated, dried over Na2SO4, and concentrated in vacuo. Product 3 was used in the next step without further purification (yield = 550 mg, 88%).

[0039] Characteristic NMR spectra 1 1H NMR (500 MHz, CDCl3): 9.01 (d, J = 8.8 Hz, 1H), 8.22 (dd, J1 = 8.2 Hz, J2 = 5.7 Hz, 1H), 8.17 (d, J = 8.3 Hz, 1H), 7.68 (m, 1H), 7.61 (t, J = 7.6 Hz, 1H), 7.16 (dd, J1 = 9.7 Hz, J2 = 8.3 Hz, 1H), 4.00 (s, 3H) ppm; 13 13C NMR (125 MHz, CDCl3): 167.0, 161.6 (d, J = 258 Hz), 133.2 (d, J = 5.4 Hz), 131.3 (d, J = 9.8 Hz), 128.7, 126.4 (d, J = 1.8 Hz), 125.9 (d, J = 2.1 Hz), 123.8 (d, J = 15.5 Hz), 122.9 (d, J = 4.3 Hz), 120.8 (d, J = 6.4 Hz), 108.3 (d, J = 20.7 Hz), 52.1 ppm.

[0040] HRMS (Orbitrap ESI): [M + H] + C 12 H 10 Calculated for C11H9FO2, 205.0659; found, 205.0658.

[0041] Step 2 - Synthesis of (4-fluoronaphthalen-1-yl)methanol (4)

[0042]

Chem.

[0043] To a stirred solution of ester 3 (550 mg, 2.70 mmol) in dry THF (10 mL) was added dropwise DIBAL-H (1.0 M solution in hexanes, 6.75 mL, 6.75 mmol) at -78 °C under an argon atmosphere. The solution was warmed to room temperature and stirred for 1 h. Complete consumption of the starting ester was confirmed by tlc analysis. The solution was then poured directly into saturated aqueous sodium potassium tartrate (10 mL) and stirred at room temperature for 1 h. The layers were separated and the aqueous layer was extracted with Et2O (2 × 10 mL). The combined organic layers were dried over Na2SO4 and concentrated in vacuo. Product 4 was used in the next step without further purification (yield = 456 mg, 96%).

[0044] Characteristic NMR spectra 1 1H NMR (500 MHz, CDCl3): δ 8.13 (m, 1H), 7.98 (m, 1H), 7.54 (m, 2H), 7.29 (dd, J1 = 7.8 Hz, J2 = 5.5 Hz, 1H), 7.02 (dd, J1 = 10.4 Hz, J2 = 7.8 Hz, 1H), 4.91 (s, 2H), 3.00 (brs, 1H) ppm; 13 13C NMR (125 MHz, CDCl3): δ 158.6 (d, J = 250.4 Hz), 132.3 (d, J = 4.7 Hz), 132.0 (d, J = 4.1 Hz), 127.0, 126.0 (d, J = 1.5 Hz), 125.1 (d, J = 8.6 Hz), 123.8 (d, J = 16.2 Hz), 123.5 (d, J = 2.7 Hz), 121.0 (d, J = 5.6 Hz), 108.5 (d, J = 19.8 Hz), 62.7 ppm.

[0045] HRMS (Orbitrap ESI): [M + H]+ + C 11 H 10 Calculated for C11H8FO, 177.0710; found, 177.0712.

[0046] Step 3 - Synthesis of 1-(bromomethyl)-4-fluoronaphthalene (5)

[0047]

Chem.

[0048] To a stirred solution of alcohol 4 (456 mg, 2.59 mmol) in dry Et2O (11 mL) was added dropwise PBr3 (110 μL, 1.16 mmol) at 0 °C under an argon atmosphere. The solution was warmed to room temperature and stirred for 30 min. Complete consumption of the starting alcohol was confirmed by tlc analysis. The solution was cooled to 0 °C using an ice bath and saturated aqueous NaHCO3 solution (10 mL) was added dropwise. The layers were separated and the aqueous layer was extracted with Et2O (2 × 10 mL). The combined organic layers were dried over Na2SO4 and concentrated in vacuo. Product 5 was used in the next step without further purification (yield = 530 mg, 86%).

[0049] Characteristic NMR spectra 1 H NMR (500 MHz, CDCl3): δ 8.18 (d, J = 8.6 Hz, 1H), 8.15 (d, J = 8.6 Hz, 1H), 7.69 (m, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.48 (dd, J1 = 7.8 Hz, J2 = 5.4 Hz, 1H), 7.08 (dd, J1 = 10.1 Hz, J2 = 7.8 Hz, 1H), 4.93 (s, 2H) ppm; 13 C NMR (125 MHz, CDCl3): δ 159.3 (d, J = 253.4 Hz), 132.3 (d, J = 5.0 Hz), 129.2 (d, J = 4.5 Hz), 127.7 (d, J = 9.0 Hz), 127.5, 126.5 (d, J = 1.5 Hz), 124.2 (d, J = 16.4 Hz), 123.8 (d, J = 2.6 Hz), 121.3 (d, J = 5.6 Hz), 108.9 (d, J = 20.1 Hz), 31.2 ppm.

[0050] Step 4 - Synthesis of 2-((4-fluoronaphthalen-1-yl)methyl)-5-methylfuran (6)

[0051]

Chemical formula

[0052] To a solution of 2-methylfuran (800 μL, 8.92 mmol) in anhydrous THF (12 mL) was added dropwise a solution of n-BuLi (1.6 M in hexane, 4.2 mL, 6.72 mmol) under an argon atmosphere at 0 °C. The solution was stirred for an additional 30 minutes at the same temperature. Then, a solution of compound 5 (530 mg, 2.23 mmol) in anhydrous THF (4 mL) was slowly added at 0 °C. The solution was then warmed to room temperature and stirred for 30 minutes. After completion of the reaction as indicated by tlc analysis (30 minutes), the reaction was quenched with a saturated aqueous solution of NH4Cl (10 mL), and the resulting mixture was extracted with Et2O (20 mL). The organic layer was separated, dried over Na2SO4, and concentrated in vacuo. The product was purified by flash column chromatography (silica gel, petroleum ether; EtOAc = 40:1) to give 6 as a pale yellow oil (yield = 487 mg, 91%).

[0053] Characteristic NMR spectra 1 1H NMR (500 MHz, CDCl3): δ 8.14 (m, 1H), 8.03 (m, 1H), 7.55 (m, 2H), 7.26 (m, 1H), 7.08 (dd, J1 = 10.3 Hz, J2 = 7.8 Hz, 1H), 5.84 (d, J = 2.7 Hz, 1H), 5.76 (d, J = 2.7 Hz, 1H), 4.32 (s, 2H), 2.26 (s, 3H) ppm; 13 13C NMR (125 MHz, CDCl3): δ 158.0 (d, J = 249.1 Hz), 152.0, 150.8, 133.0 (d, J = 4.2 Hz), 130.1 (d, J = 4.5 Hz), 126.8, 126.4 (d, J = 8.2 Hz), 125.8, 124.1 (d, J = 2.5 Hz), 124.0 (d, J = 17.0 Hz), 121.1 (d, J = 5.5 Hz), 108.9 (d, J = 19.5 Hz), 107.3, 106.1, 31.6, 13.5 ppm.

[0054] HRMS (Orbitrap ESI); [M+H]+ + C 16 H 14 Calculated for C15H13FO, 241.1023; found, 241.1024.

[0055] Synthesis of 5 - 5-(4-fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-en-1-one (7, enone-2)

[0056] [Chemical formula]

[0057] Furan 6 (120 mg, 0.5 mmol) was dissolved in MeOH (5 mL, 0.1 M) containing a catalytic amount (7.3 mg, 1.5% mol) of rose bengal as a photosensitizer. The solution was cooled in an ice bath. While irradiating the solution with a xenon Variac Eimac Cermax 300W lamp, oxygen was gently bubbled through the solution. After completion of the reaction (5 minutes, indicated by tlc), the solution was warmed to room temperature and Me2S (146 μL, 2 mmol) was added. After completion of the reduction (40 minutes) indicated by tlc analysis, Et3N (21 μl, 0.15 mmol) was added and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was observed by tlc analysis and 1 1H-NMR. After completion of the reaction (30 minutes), the solvent was removed in vacuo and the residue was purified by flash column chromatography (silica gel, petroleum ether:EtOAc = 7:1 → 4:1) to give 7 as a white solid (yield = 85 mg, 66%). To scale up the process, the 5-minute irradiation step was repeated 10 times on the above scale, the photo-oxidized solutions were combined, and then treated with Me2S and Et3N.

[0058] Characteristic NMR spectra 1 1H NMR (500 MHz, CDCl3): δ 8.20 (brs, 1H), 8.15 (m, 1H), 7.56 (m, 2H), 7.50 (d, J = 5.6 Hz, 1H), 7.10 (dd, J1 = 9.8 Hz, J2 = 8.3 Hz, 1H), 6.97 (brs, 1H), 6.32 (d, J = 5.6 Hz, 1H), 4.50 (s, 1H), 2.59 (s, 1H), 0.95 (s, 3H) ppm; 1313C NMR (125 MHz, CDCl3): 207.3, 165.9, 158.1 (d, J = 250.3 Hz), 134.8, 132.3, 128.5 (d, J = 3.9 Hz), 127.4, 126.6 (d, J = 8.1 Hz), 126.1, 124.1, 124.0 (d, J = 17.0 Hz), 121.2 (d, J = 4.7 Hz), 108.9 (d, J = 20.0 Hz), 80.9, 60.1, 25.2 ppm.

[0059] HRMS (Orbitrap ESI): [M+H] + 13C 16 1H 14 Calculated value of C17H13FO2, 257.0972; found, 257.0969.

[0060] Experiments to demonstrate the cytotoxic effects of enone-1 and enone-2 on malignant cells This experiment aims to demonstrate the cytotoxic effects of 4-hydroxy-4-methyl-5-(naphthalen-2-yl)cyclopent-2-enone (enone-1) and 5-(4-fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-enone (enone-2), the final products of chemical synthesis, against ovarian cancer, colorectal cancer, cervical cancer, hepatocellular carcinoma, lung cancer, bladder cancer, breast cancer, melanoma, lymphoma, and myeloma malignant cells.

[0061] Specifically, enone-1 and enone-2 were tested for cytotoxic activity in 17 cell lines representing a wide range of human tumors. They included human colorectal cancer (Caco2, RKO, HCT15, HT29), liver (HepG2), lung (A549), ovarian cancer (A2780, 2780CP, AGE60, SKOV3), undifferentiated large cell lymphoma (Mac1 and Mac2a), breast cancer (MDA-MB-231), cervical (HeLa), bladder cancer (VM-CUB-1), melanoma (BRO), and multiple myeloma (MR20).

[0062] Typically, 7000 - 8000 cancer cells were seeded into 96 - well flat - bottom plates in standard growth medium. After 16 hours, the cells were treated with increasing concentrations of enone - 1, enone - 2, or cisplatin, a chemotherapeutic agent established as a comparison, for 48 hours. Then, cell viability was evaluated by using the MTT conversion assay [1]. Each assay was performed in triplicate and repeated in 2 - 3 independent experiments. Specifically, to measure cell viability, 20 μl of 10 mg / ml MTT (3 - (4,4 - dimethylthiazol - 2 - yl) - 2,5 - diphenyltetrazolium bromide; Sigma) was added to each well, and the plates were incubated at 37 °C in 5% CO2 for 3 hours. The supernatant was removed, and the formed crystals were dissolved in 200 μl of dimethyl sulfoxide. Then, the plates were analyzed at 450 nm with a plate reader. Growth inhibition (decrease in viability) was calculated by expressing the difference in optical density between the treated wells and the control wells as a percentage of the control.

[0063] Representative results of the MTT conversion assay are shown in Figure 1, and aggregated data from 3 independent experiments are shown in Table 1. Both enone - 1 and enone - 2 produced significant cytotoxic effects that replaced the cytotoxic effect of cisplatin. In each cell line tested, there was little difference in the 50% inhibitory concentration (IC50) of enone - 1 and enone - 2. The IC50 values ranged from approximately 0.05 μM in MR20 multiple myeloma cells (the most sensitive) to 6.2 μM in HeLa cervical cancer cells (less sensitive). For comparison, the IC50 value of cisplatin was approximately 5.8 μM for MR20 and 31 μM for HeLa cells. Thus, enone 1 and enone 2 showed 2.5 - 50 - fold higher toxicity than cisplatin in the 48 - hour exposure assay measured by MTT conversion, depending on the cell line.

[0064]

Table 1

[0065] The effects of enone-1 and enone-2 on cancer cells were confirmed using a second method, namely the PrestoBlue assay (PrestoBlue™ Cell Viability Reagent, Thermo Fisher, catalog number: A13261), a cell-permeable resazurin-based solution that functions as an indicator of cell survival. A2780 ovarian cancer cells were plated as described above for the MTT assay, and enone agents were added at increasing concentrations as shown in Figure 2. Cell survival was determined 72 hours after addition of PrestoBlue according to the manufacturer's (Thermo Fisher) instructions. For comparison, the effects of two platinum-based chemotherapeutic agents, cisplatin and oxaliplatin, were tested in parallel.

[0066] All agents showed a concentration-dependent decrease in PrestoBlue fluorescence, which indicates a decrease in A2780 survival (Figure 2). The IC50 values of enone-1 and enone-2 using this experimental setup were 0.20 μM (±0.06) and 0.12 μM (±0.07), respectively, from experiments with n = 3, the IC50 of cisplatin was 1.4 μM (±0.12), and the IC50 of oxaliplatin was 0.27 μM (±0.03). Thus, enone-1 and enone-2 induced approximately 8- to 13-fold higher cytotoxicity and 1.25- to 2-fold higher cancer cell killing efficacy in A2780 cells compared to cisplatin and higher than oxaliplatin as measured by the PrestoBlue assay.

[0067] The long-term effects of enone-2 on tumor cell survival were examined using a clonogenic assay that evaluates the ability of single cells to survive and replicate to form colonies. A2780 ovarian cancer cells were treated with 2.5 μM or 5.0 μM enone-2 or cisplatin for 12 hours, then washed with PBS to remove excess drug, detached by trypsinization, counted, and seeded into 6-well tissue culture plates (Costar) at 600 cells per well. Colonies formed on day 10 were visualized and counted by staining with crystal violet after cell fixation. Under these conditions, the number of colonies formed by A2780 cells treated with enone-2 was lower than that of the equivalent dose of cisplatin (Figure 3).

[0068] Therefore, enone-1 and enone-2 exert potent cytotoxic effects in several cancer cell lines and substitute for the cytotoxic effects of the established chemotherapeutic agents cisplatin and oxaliplatin.

[0069] Experiments demonstrating that enone-1 and enone-2 inhibit cancer cell proliferation by inhibiting cell cycle progression in the G2 / M phase and by decreasing the expression of DNA replication licensing factors. The effect of enone-2 on cell cycle progression was tested to determine whether it causes cell cycle arrest in addition to inducing cell death. For this purpose, A2780 ovarian cancer cells were treated with 2.5 μM or 5 μM enone-2 for 24 hours, the cells were detached, fixed in 1% neutral buffered formalin, and stained with 5 μg / ml propidium iodide. Next, the cell suspension was analyzed for fluorescence intensity (DNA content) by quantitative flow cytometry. As shown in Figure 4A, enone-2 caused a concentration-dependent accumulation of cells in the G2 / M phase of the cell cycle.

[0070] Replication licensing factors (RLFs) such as CDC6, MCM2-MCM7, ORC, and CDT1 form the origin of DNA replication in the G1 phase of the cell cycle and initiate DNA replication during the S phase. CDC6 is also involved in the S-M checkpoint mechanism that ensures that the entire genome is replicated only once per cell division. Cancer cells often exhibit deregulated expression of RLFs. For example, CDC6 is overexpressed in colorectal and ovarian cancers and is associated with poor patient prognosis [8, 9]. Higher levels of CDC6 are also observed in cisplatin-resistant tumor cell lines and contribute to resistance to both cisplatin and oxaliplatin [10, 11]. Because of these characteristics, RLFs are considered novel targets for cancer therapy

[12] . Enon-2 was found to decrease the important RLF CDC6 in a time-dependent manner at both the RNA and protein levels (Figures 4B and 4C), and also decrease CDT1 RNA levels. Therefore, the inventors claim that enon targets the molecular pathway that results in decreased expression of CDC6 and other RLFs and induces arrest at the G2 / M phase of the cell cycle. Therefore, the anticancer effects of enon-1 and enon-2 may be particularly relevant to cancers that overexpress RLFs.

[0071] Experiment to prove that enon-2 suppresses tumor growth in vivo. The in vivo antitumor effect of enon was determined by xenotransplantation of MB49 mouse bladder cancer cells in syngeneic immunocompetent C57BL / 6 female mice.

[0072] First, the effects of enon-1 and enon-2 on the viability of MB49 cells were verified by the MTT assay performed as described in Figure 1. As shown in Figure 5A, the IC50 of enon-2 was found to be 0.6 μM (±0.1), 1.4 μM (±0.2) for enon-1, and 3.8 μM (±0.3) for cisplatin.

[0073] Based on this, the in vivo antitumor activity of enon-2 was tested. Female mice were given 10 6MB49 cells were subcutaneously implanted. By the 7th day, tumors with a size of approximately 40 - 60 mm 2 were formed. Mice were randomly divided into two groups of five each, and on the 7th, 10th, 14th, and 20th days, 50 μl of enone - 2 was injected into the tumors to achieve a drug dosage of 5 mg / kg. The tumor size was observed using digital calipers, and was expressed in mm 2 by multiplying the length (longest diameter) of the tumor by the width (shortest diameter perpendicular to the length). As shown in Figure 5, enone - 2 halted tumor growth, while the MB49 tumors that received the vehicle control (VC, 50 μl of physiological saline) continued to grow.

[0074] Experiment to prove that enone - 1 and enone - 2 overcome resistance to cisplatin in vitro. Resistance to platinum - based chemotherapy is a major problem in oncology that affects treatment options and limits patient survival. This is associated with several types of malignant tumors such as ovarian cancer where platinum resistance frequently occurs. Since enone - 1 and enone - 2 show efficacy in reducing the survival of A2780 ovarian cancer cells, these drugs were also tested in 2780CP (a cisplatin - resistant derivative of A2780)

[13] . These cells show approximately 7 - fold lower sensitivity to cisplatin compared to A2780 cells (Figure 6 and Figure 1D). Nevertheless, 2780CP cells were found to have approximately the same sensitivity to enone - 1 and enone - 2 as cisplatin - sensitive A2780 cells by the MTT conversion assay.

[0075] Therefore, the inventors claim that cisplatin - resistant cells do not confer cross - resistance to enone, and that enone - 1 and enone - 2 may provide new treatment options in platinum - resistant tumors.

[0076] Experiment to prove that enone - 2 is an inducer of ribosomal biogenesis stress. Gene expression profiling of A2780 ovarian cancer cells exposed to 2.5 μM enon-2 for 6 or 14 hours revealed activation of the p53 pathway by upregulation of several p53 target genes (Figure 7). Activation of p53 occurs in response to DNA damage (so-called "DNA damage response", DDR) or ribosomal stress (RS), and transmits signals that arrest the cell cycle and induce cell death.

[0077] The effects of enon-2 on p53 transcription and p53-dependent transcription of cell cycle and cell death-related genes were confirmed by immunoblotting for p53 and by reverse transcription qPCR (RT-qPCR) for mdm2, p21, bax, and bbc3 / puma, an established p53 target gene. As shown in Figure 8, treatment of A2780 ovarian cancer cells or AGE60 ovarian cancer cells with 2.5 μM enon-2 resulted in a steady accumulation of p53, while cisplatin at an equivalent concentration was less effective, consistent with their differential effects on cell survival. A2780 responded to enon-2 by time-dependent upregulation of mdm2, p21, bax, and bbc3 / puma mRNA measured by RT-qPCR. Enon-1 was also found to induce p53 gene expression and p53-dependent gene expression in A2780 cells (Figure 8).

[0078] Anticancer agents can induce p53 using different pathways. For example, cisplatin induces p53 mainly via DDR, while oxaliplatin activates p53 via RS

[14] . Due to these differences, it has been proposed that cisplatin and oxaliplatin "should be used in a mechanism-targeted manner for cancer treatment"

[14] , and that drugs that induce RS may be particularly beneficial for tumors with "translation dependence".

[0079] To determine whether enon-mediated p53 activation is mediated by DDR or RS, protein lysates isolated from A2780 cells cultured for 6 hours with increasing concentrations of either enon-2 or cisplatin were analyzed for yH2Ax, a marker of DDR. As shown in the representative immunoblot of Figure 9A, cisplatin induced yH2Ax, while enon-2 had no effect compared to the untreated control. However, enon-2 steadily induced p53 accumulation under the same conditions (Figure 9A). This finding suggests that enon-mediated p53 activation is not involved in the DDR pathway.

[0080] To further validate this observation, A2780 cells were transiently transfected with siRNAs targeting Chek2, which is required for DDR-induced p53, or Rpl11, which is required for RS-mediated p53 activation

[14] . The efficacy and specificity of the knockdown were confirmed by RT-qPCR (Figure 9B). As expected

[14] , Chek2 knockdown improved cisplatin-induced p53 activation, and depletion of Rpl11 decreased oxaliplatin-induced p53 accumulation. Importantly, only Rpl11 knockdown affected enon-2-mediated p53 activation (Figures 9C and 9D). This DDR-independent mechanism of p53 activation is consistent with the observation that enon-2 does not show cross-resistance with cisplatin in apoptosis.

[0081] Therefore, the inventors claim that enon-1 and enon-2 are inducers of the p53 pathway mediated by the ribosome stress pathway. Therefore, enon may be particularly beneficial for "translation-dependent" tumors.

[0082] An experiment demonstrating that enon-2 synergizes with a proteasome inhibitor in inducing cancer cell death in vitro. Gene expression profiling of A2780 ovarian cancer cells treated with 2.5 μM enon-2 for 6 hours, followed by bioinformatics analysis, demonstrated enrichment of upregulated transcripts related to the proteasome-mediated ubiquitin-dependent protein catabolic process. This group included gene transcripts, namely ZFAT2 A, KCTD10, MDM2, PLK2, DDIT3, DNAJB9, PSMD6, RYBP, PSMD10, SPSB3, PSMB8, PELI1, RNF103, DNAJB2, UBE2H, ATXN3, PSEN1, RCHY1, UBR1, ANAPC16, JKAMP, DDA1, PSMA6, TMUB2, TRIM72, SPOPL, UBE2J1, ARIH1, NPLOC4, CDC26, USP19, UFD1, TNFAIP1, PSMB6, DERL1, UBE2V2, SELENOS, PSMA1, UBE2.

[0083] Proteasome inhibitors have been reported to enhance platinum-induced toxicity in cisplatin-resistant ovarian cancer cells

[15] . Significant enrichment of transcripts related to proteasome activity in enon-treated cells suggested that proteasome inhibitors might also enhance enon-2-induced cytotoxicity. To address this hypothesis, 2780CP ovarian cancer cells were pretreated with different concentrations of the proteasome inhibitor bortezomib (a clinically used drug) for 2 hours and then co-cultured with enon-2 for 48 hours. The results demonstrated an increase in cytotoxicity compared to each drug alone (Figure 10).

[0084] Therefore, the inventors claim that enon can be used in combination with proteasome inhibitors for cancer treatment. This may be particularly relevant to ovarian cancer and multiple myeloma where bortezomib is clinically used.

[0085] References 1. Gordon, A.N.; Fleagle, J.T.; Guthrie, D.; Parkin, D.E.; Gore, M.E.; Lacave, A.J. Recurrent epithelial ovarian carcinoma: a randomized phase III study of pegylated liposomal doxorubicin versus topotecan. J Clin Oncol 2001, 19, 3312 - 3322, doi:10.1200 / JCO.2001.19.14.3312。 2. Rose, P.G.; Yao, M.; Chambers, L.M.; Mahdi, H.; DeBernardo, R.; Michener, C.M.; AlHilli, M.; Ricci, S.; Vargas, R. PARP inhibitors decrease response to subsequent platinum - based chemotherapy in patients with BRCA mutated ovarian cancer. Anticancer Drugs 2021, 32, 1086 - 1092, doi:10.1097 / CAD.0000000000001219。 3. Solimini, N.L.; Luo, J.; Elledge, S.J. Non - oncogene addiction and the stress phenotype of cancer cells. Cell 2007, 130, 986 - 988, doi:10.1016 / j.cell.2007.09.007。 4. Graff, J.R.; Konicek, B.W.; Vincent, T.M.; Lynch, R.L.; Monteith, D.; Weir, S.N.; Schwier, P.; Capen, A.; Goode, R.L.; Dowless, M.S.; et al. Therapeutic suppression of translation initiation factor elF4E expression reduces tumor growth without toxicity. J Clin Invest 2007, 117, 2638 - 2648, doi:10.1172 / JCI32044。 5. Wang, C.; Zhang, J.; Yin, J.; Gan, Y.; Xu, S.; Gu, Y.; Huang, W. Alternative approaches to target Myc for cancer treatment. Signal Transduct Target Ther 2021, 6, 117, doi:10.1038 / S41392 - 021 - 00500 - y。 6. Kalaitzakis, D.; Triantafyllakis, M.; Alexopoulou, I.; Sofiadis, M.; Vassilikogiannakis, G. One - pot transformation of simple furans into 4 - hydroxy - 2 - cyclopentenones in water. Angew Chem Int Ed Engl 2014, 53, 13201 - 13205, doi:10.1002 / anie.201407477。 7. Ioannou, G.I.; Montagnon, T.; Kalaitzakis, D.; Pergantis, S.A.; Vassilikogiannakis, G. Synthesis of cyclopent - 2 - enones from furans using a nebulizer - based continuous flow photoreactor. Org Biomol Chem 2017, 15 10151 - 10155, doi:10.1039 / c7ob02557b。 8. Yang, C.; Xie, N.; Luo, Z.; Ruan, X.; Zhang, Y.; Wang, W.; Huang, Y. The Effect of High CDC6 Levels on Predicting Poor Prognosis in Colorectal Cancer. Chemotherapy 2022, 67, 47 - 56, doi:10.1159 / 000519913。 9. Deng, Y.; Jiang, L.; Wang, Y.; Xi, Q.; Zhong, J.; Liu, J.; Yang, S.; Liu, R.; Wang, J.; Huang, M.; et al. High expression of CDC6 is associated with accelerated cell proliferation and poor prognosis of epithelial ovarian cancer. Pathol Res Pract 2016, 212, 239 - 246, doi:10.1016 / j.prp.2015.09.014。 10. Chen, S.; Chen, X.; Xie, G.; He, Y.; Yan, D.; Zheng, D.; Li, S.; Fu, X.; Li, Y.; Pang, X.; et al. Cdc6 contributes to cisplatin - resistance by activation of ATR - Chk1 pathway in bladder cancer cells. Oncotarget 2016, 7, 40362 - 40376, doi:10.18632 / oncotarget.9616. 11. Cai, J.; Wang, H.; Jiao, X.; Huang, R.; Qin, Q.; Zhang, J.; Chen, H.; Feng, D.; Tian, X.; Wang, H. The RNA - Binding Protein HuR Confers Oxaliplatin Resistance of Colorectal Cancer By Upregulating CDC6. Mol Cancer Ther 2019, 18, 1243 - 1254, doi:10.1158 / 1535 - 7163.MCT - 18 - 0945。 12. Lim, N.; Townsend, P. A. Cdc6 as a novel target in cancer: Oncogenic potential, senescence and subcellular localisation. Int J Cancer 2020, 147, 1528 - 1534, doi:10.1002 / ijc.32900。 13. Eliopoulos, A. G.; Kerr, D. J.; Herod, J.; Hodgkins, L.; Krajewski, S.; Reed, J. C.; Young, L. S. The control of apoptosis and drug resistance in ovarian cancer: influence of p53 and Bcl-2. Oncogene 1995, 11, 1217 - 1228。 14. Bruno, P. M.; Liu, Y.; Park, G. Y.; Murai, J.; Koch, C. E.; Eisen, T. J.; Pritchard, J. R.; Pommier, Y.; Lippard, S. J.; Hemann, M. T. A subset of platinum-containing chemotherapeutic agents kills cells by inducing ribosome biogenesis stress. Nat Med 2017, 23, 461 - 471, doi:10.1038 / nm.4291。 15. Huang, W.; Zhou, Q.; Yuan, X.; Ge, Z. M.; Ran, F. X.; Yang, H. Y.; Qiang, G. L.; Li, R. T.; Cui, J. R. Proteasome Inhibitor YSY01A Enhances Cisplatin Cytotoxicity in Cisplatin-Resistant Human Ovarian Cancer Cells. J Cancer 2016, 7, 1133 - 1141, doi:10.7150 / jca.14519。

Claims

1. A composition for use as a medicament for the treatment of cancer in a mammal, comprising a 5-substituted 4-hydroxycyclopent-2-en-1-one derivative, wherein the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative has the structure: 【Chemical Formula 1】 having the formula, wherein R 1 is phenyl, 2-naphthyl, 1-naphthyl, -CO 2 R, alkyl, and R 2 is -H, alkyl, alkenyl, benzyl, R 3 is -H, alkyl, and R 4 A composition characterized in that R is -H or alkyl.

2. A composition for use as a medicament for the treatment of cancer in a mammal, comprising the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to claim 1, wherein the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative has the structure: [Chemical Formula 2] having the formula, wherein R 1 is phenyl, 4-fluorophenyl, naphthalen-2-yl, 4-fluoronaphthalen-1-yl, CO 2 Et, CO 2 Me, n-butyl, and R 2 is H, methyl, n-C 16 H 33 , benzyl, n-pentyl, n-hexyl, pent-4-enyl, and R 3 is H, methyl, and R 4 is H, methyl, and Preferably, the composition has a trans stereochemistry such that the 4-hydroxy group is trans to the substituent at the 5-position.

3. A composition for use as a medicament for the treatment of cancer in a mammal, comprising the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 1 or 2, wherein the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative is 5-(4-fluoronaphthalen-1-yl)-4-hydroxy-4-methylcyclopent-2-en-1-one or 4-hydroxy-4-methyl-5-(naphthalen-2-yl)cyclopent-2-en-1-one, and preferably the composition has a trans stereochemistry such that the 4-hydroxy group is trans to the substituent at the 5-position.

4. A composition for use as a medicament for the treatment of cancer in a mammal, comprising the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 1 to 3, wherein the composition further comprises a delivery vehicle, and preferably the delivery vehicle is a liposome.

5. A composition for use for the treatment of cancer in a mammal, comprising the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 1 to 4, wherein the composition further comprises a proteasome inhibitor, and preferably the proteasome inhibitor is bortezomib.

6. A composition for use for the treatment of cancer in a mammal, comprising the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 1 to 5, A composition, wherein the cancer is resistant to platinum-based chemotherapy, preferably cisplatin treatment and / or oxaliplatin. **Claim 7** A composition for use in the treatment of cancer in a mammal, comprising a 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 1 to 6, wherein the cancer exhibits translation dependence. **Claim 8** A composition for use in the treatment of cancer in a mammal, comprising a 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 1 to 7, wherein the cancer is selected from the group consisting of basal cell carcinoma, medulloblastoma, liver cancer, rhabdomyosarcoma, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, colon cancer, breast cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, myeloma, bladder cancer, kidney cancer or ureteral cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) neoplasm, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma, or a combination of one or more of the aforementioned cancers. **Claim 9** A composition for the treatment of cancer according to any one of claims 1 to 8, administered to a subject, wherein the pharmaceutical composition comprises a compound according to any one of claims 1 to 8, or a tautomer or pharmaceutically acceptable salt thereof, and a delivery vehicle such as a liposome that enhances the stability of the compound and / or improves the pharmacokinetics and toxicity profile in organs. **Claim 10** A 5-substituted 4-hydroxycyclopent-2-en-1-one derivative used in the composition according to any one of claims 1 to 9, having the following formula [Chemical 3] A 5-substituted 4-hydroxycyclopent-2-en-1-one derivative having the formula. **Claim 11** A 5-substituted 4-hydroxycyclopent-2-en-1-one derivative used in the composition according to any one of claims 1 to 9, having the following formula 【Chemical Formula 4】 A 5-substituted 4-hydroxycyclopent-2-en-1-one derivative having the formula. **Claim 12** The 4-hydroxy group is trans to the substituent at the 5-position, the 5-substituted 4-hydroxycyclopent-2-en-1-one derivative according to any one of claims 10 or 11.

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