Pharmaceutical composition for combined administration with anticancer drug containing phthalazinone derivative

Abstract

The present application provides a combination of a pharmaceutical composition for co-administration, comprising (i) a pharmaceutical composition comprising a phthalazinone derivative, and (ii) a pharmaceutical composition comprising at least one anti-cancer agent. The combination of the present invention shows a superior synergistic effect compared to the administration of an existing anti-cancer agent alone.

Description

[Technical field]

[0001] [CROSS REFERENCE TO RELATED APPLICATIONS] This application claims priority to U.S. Provisional Patent Application No. 63 / 302,834, filed January 25, 2022, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to a pharmaceutical composition for combined administration containing a phthalazinone derivative used for the prevention or treatment of cancer, and more particularly to a pharmaceutical composition for combined administration containing a phthalazinone derivative and an anticancer agent.

[0003] The present invention also relates to a pharmaceutical composition for combined administration comprising a phthalazinone derivative and irinotecan and / or oxaliplatin, which is used for the prevention or treatment of cancer.

[0004] The present invention also relates to methods of treating cancer in a patient comprising administering irinotecan and / or oxaliplatin in combination with a phthalazinone derivative for the prevention or treatment of cancer. [Background technology]

[0005] Although existing therapies (radiotherapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, etc.) provide complete remission (complete response) to some cancer patients, most cancer patients show moderate response or non-response, and a certain number of cancer patients still show disease progression. Therefore, attempts to treat some patients with advanced cancer using such existing therapies and maintenance therapies are limited.

[0006] Recently, to address the heterogeneity and emergence of cancer cells with treatment resistance within a cancer cell population, a therapeutic method of administering PARP inhibitors (PARP-1, PARP-2, and PARP-3 (collectively referred to as PARP)) that inhibit poly(ADP-ribose) polymerase has been successfully used.

[0007] PARP is an enzyme system involved in DNA repair, consisting of 18 components, which are located in the cell nucleus and are key enzymes in the DNA repair mechanism of human cells when single-stranded DNA in cancer cells is damaged.

[0008] Such PARP inhibitors induce and increase intracellular DNA damage via the homologous recombination restoration process. In cancer cells that already have homologous recombination deficiency (HRD), PARP inhibitors block the action of PARP to accumulate DNA damage in cancer cells and induce cell death.

[0009] Examples of PARP inhibitors include olaparib (AZD-2281), rucaparib (AG-014699), niraparib (MK-4827), talazoparib (BMN-673), veliparib (ATB-888), fluzoparib (SHR-3162), pamiparib (BGN-290), senaparib (IMP-4297), stenoparib (2X-121) and AZD-5305. In particular, olaparib, as a potent inhibitor of human PARP, inhibits the in vitro growth of certain tumor cell lines and the in vivo growth of tumors, either as a monotherapy or in combination with established chemotherapy (Cancers 2020, 12(2), 334; doi: 10.3390 / cancers12020334).

[0010] However, when Olaparib is used as a therapeutic agent, there is a problem that cancer cells that have acquired resistance to Olaparib appear in vivo, resulting in a reduced therapeutic effect. Therefore, there is an urgent need to develop a new therapeutic agent that can effectively treat cancers that are resistant to specific PARP inhibitors such as Olaparib.

[0011] Although monotherapy using specific PARP inhibitors alone has shown good efficacy and safety profile in clinical trials, major limitations include the need for HRD and the rapid development of resistance. Many tumors that initially respond to treatment using PARP inhibitors eventually relapse through compensatory mutations that restore homologous recombination activity or stimulate the activity of alternative repair pathways. Therefore, the use of specific PARP inhibitors is limited to certain tumor types and cannot be used to treat all cancers.

[0012] U.S. Patent No. 9,682,973 (Patent Document 1), the entire contents of which are incorporated herein by reference, discloses a PARP inhibitor compound represented by Chemical Formula 1, which is a phthalazinone derivative having anticancer activity, or a pharma- ceutically acceptable salt thereof. In particular, the hydrochloride form of the compound represented by Chemical Formula 1 is one of a group of promising therapeutic drug candidates as an anticancer agent. [ka] [Prior art documents] [Patent documents]

[0013] [Patent Document 1] U.S. Patent No. 9,682,973 Summary of the Invention [Problem to be solved by the invention]

[0014] An object of the present invention is to provide a pharmaceutical composition for combined administration comprising a phthalazinone derivative having excellent activity as a poly(ADP-ribose) polymerase (PARP) inhibitor and at least one anticancer agent.

[0015] An object of the present invention is to provide a pharmaceutical composition for co-administration comprising a compound represented by Chemical Formula 1, which is a phthalazinone derivative, or a pharma- ceutically acceptable salt thereof.

[0016] The object of the present invention is to provide a kit comprising a phthalazinone derivative having excellent activity as a PARP inhibitor and at least one anticancer agent capable of being administered in combination, which is a combined preparation for simultaneous, separate or sequential use in cancer treatment.

[0017] The object of the present invention is to provide a method for treating cancer, which comprises administering a PARP inhibitor and at least one anti-cancer agent, either simultaneously or sequentially, in combination for the treatment of cancer, including but not limited to those associated with homologous recombination deficiency (HRD).

[0018] Furthermore, the present invention provides a synergistic therapeutic effect against tumors by administering the compound represented by Chemical Formula 1 or a pharma- ceutically acceptable salt thereof in combination with an anticancer agent. [Means for solving the problem]

[0019] One aspect of the present invention provides a pharmaceutical composition for combined administration comprising, as active ingredients, (i) a compound represented by Chemical Formula 1 {4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl)-4-fluorobenzyl]phthalazin-1(2H)-one}, or a pharma- ceutically acceptable salt thereof, and (ii) at least one anticancer agent: [ka] .

[0020] In another embodiment of the present invention, there is provided a combination of pharmaceutical compositions for combined administration for preventing or treating cancer, comprising: (i) a first pharmaceutical composition comprising, as an active ingredient, a compound represented by the following chemical formula 1, or a pharma- ceutically acceptable salt thereof; [ka] and (ii) a second pharmaceutical composition comprising, as an active ingredient, at least one anti-cancer agent.

[0021] In the present invention, the compound represented by Chemical Formula 1 is also a type of phthalazinone derivative.

[0022] As used herein, a "compound represented by Chemical Formula 1" may simply be referred to as a "compound of Chemical Formula 1" or a "compound of the invention." Similarly, a "salt of a compound represented by Chemical Formula 1" may be referred to as a "salt of a compound of Chemical Formula 1" or a "salt compound of Chemical Formula 1." For example, when a compound represented by Chemical Formula 1 exists in the form of a hydrochloride salt, the compound may also be referred to as a "hydrochloride salt of a compound of Chemical Formula 1" or a "hydrochloride compound of Chemical Formula 1." Such salt forms include inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, etc.), organic carboxylic acids (e.g., acetic acid, haloacetic acid such as trifluoroacetic acid, propionic acid, maleic acid, succinic acid, malic acid, citric acid, tartaric acid, salicylic acid, etc.), acidic sugar acids (e.g., glucuronic acid, galacturonic acid, gluconic acid, ascorbic acid, etc.), acidic polysaccharides (e.g., hyaluronic acid, chondroitin sulfate, arginic acid, etc.), sulfonic acids such as chondroitin sulfate, organic sulfonic acids including sugar esters (e.g., methanesulfonic acid, p-toluenesulfonic acid, etc.), etc. Preferably, the salt form of the compound represented by Formula 1 is a hydrochloride salt.

[0023] In the present invention, the pharma- ceutically acceptable salt may be prepared by a suitable method in the art by preparing the compound represented by Chemical Formula 1 in the form of a glass base. For example, the free base may be prepared by treating the free base with an inorganic acid (e.g., hydrochloric acid, bromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, etc.), an organic acid (e.g., acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, etc.), a pyranosidyl acid (e.g., glucuronic acid, galacturonic acid, etc.), an alpha hydroxyl acid (e.g., citric acid, tartaric acid, etc.), an amino acid (e.g., aspartic acid, glutamic acid, etc.), an aromatic acid (e.g., benzoic acid, cinnamic acid, etc.), a sulfonic acid (e.g., p-toluenesulfonic acid, ethanolsulfonic acid, etc.), etc.

[0024] The compound represented by Chemical Formula 1 of the present invention or a pharma- ceutically acceptable salt thereof can be easily obtained by, for example, the method disclosed in US Pat. No. 9,682,973 or by the method therefor.

[0025] As used herein, anti-cancer drug can refer to any drug used to kill cancer cells.

[0026] In some embodiments, the anti-cancer drug in the second pharmaceutical composition is selected from the group consisting of chemotherapy drugs, targeted therapy drugs, immunotherapy drugs and hormonal therapy drugs. The type of anti-cancer drug that can be used in the composition of the present invention is not particularly limited. The anti-cancer drug can be selected according to general principles that are considered when selecting an anti-cancer drug, such as the type of cancer cells, the absorption rate of the anti-cancer drug (treatment period and administration route of the anti-cancer drug), the location of the tumor, and the size of the tumor. Specifically, the anti-cancer drugs that can be used in the present invention may be (i) chemotherapy drugs such as alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, mitotic inhibitors, or other chemotherapy drugs; (ii) targeted therapy drugs such as angiogenesis inhibitors, monoclonal antibodies, proteasome inhibitors, signal transduction inhibitors, or new lineages; (iii) immunotherapy drugs such as checkpoint inhibitors, chimeric antigen receptor (CAR) T-cell therapy, or cytokines; or (iv) hormone therapy drugs such as aromatase inhibitors, selective estrogen receptor modulators, estrogen receptor antagonists, luteinizing hormone releasing hormone agonists, antiandrogens, or CYP17 inhibitors; but are not limited thereto.

[0027] As mentioned above, in some embodiments, the chemotherapy drug is selected from the group consisting of alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, and mitotic inhibitors. In some embodiments, the targeted therapy drug is selected from the group consisting of angiogenesis inhibitors, monoclonal antibodies, proteasome inhibitors, and signal transduction inhibitors. In some embodiments, the immunotherapy drug is selected from the group consisting of checkpoint inhibitors, chimeric antigen receptor (CAR) T cell therapy, and cytokines. In some embodiments, the hormone therapy drug is selected from the group consisting of aromatase inhibitors, selective estrogen receptor modulators, estrogen receptor antagonists, luteinizing hormone releasing hormone enhancers, antiandrogens, and CYP17 inhibitors.

[0028] The anticancer drug usable in the present invention is preferably a DNA alkylating agent that is an anticancer drug belonging to the platinum group or an anticancer drug belonging to the topoisomerase inhibitor group. The anticancer drug belonging to the platinum group in the DNA alkylating agent is selected from altretamine, carboplatin, cisplatin, cyclophosphamide, dacarbazine, ifosfamide, oxaliplatin, and temozolomide, and is preferably, but not limited to, oxaliplatin. The anticancer drug belonging to the topoisomerase inhibitor class is selected from irinotecan, irinotecan liposome, topotecan, etoposide, and teniposide, and is preferably irinotecan, but is not limited thereto.

[0029] In certain embodiments, the anti-cancer drug is selected from irinotecan and oxaliplatin. In certain embodiments, the anti-cancer drug is irinotecan. In some embodiments, the anti-cancer drug is oxaliplatin.

[0030] The pharmaceutical composition of the present invention is administered in the form of a mixture of a compound represented by Chemical Formula 1 or a pharma- ceutically acceptable salt thereof and at least one anticancer agent, and the at least one anticancer agent is formulated and administered simultaneously or sequentially.

[0031] In some embodiments, the combined pharmaceutical composition is administered in the form of a mixture of the first pharmaceutical composition and the second pharmaceutical composition.

[0032] In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are each comprised in a separate dosage form.

[0033] In some embodiments, the combined pharmaceutical composition is administered in a manner in which the first pharmaceutical composition and the second pharmaceutical composition are each formulated and administered simultaneously or sequentially.

[0034] The pharmaceutical composition of the present invention is also a form for simultaneous administration of two drugs, including a mixed form in which the compound represented by Chemical Formula 1 or a pharma- ceutical acceptable salt thereof is mixed with an anticancer drug. The pharmaceutical composition of the present invention is also a form in which the compound represented by Chemical Formula 1 or a pharma- ceutical acceptable salt thereof and an anticancer drug are each formulated and administered simultaneously or sequentially. In the case of sequential administration, the order may be interchanged. According to one embodiment of the present invention, it has been confirmed that when the hydrochloride salt of the compound represented by Chemical Formula 1 is administered in combination with an anticancer drug, an anti-proliferative effect is exhibited on cancer cells in a concentration-dependent manner (see FIG. 1 and FIG. 2).

[0035] As used herein, "simultaneously" is used to mean that two formulations or compositions (eg, pharmaceutical compositions) are administered concurrently.

[0036] As used herein, "sequentially" is used to mean that the active ingredients or compositions (e.g., pharmaceutical compositions) are not administered simultaneously, but rather one after the other. Thus, "sequential" administration may allow one formulation / composition (e.g., pharmaceutical composition) to be administered within 5 minutes, 10 minutes, or hours after another formulation / composition, provided that the circulating half-life of the formulation administered first is long enough that the two formulations may be present simultaneously in therapeutically effective amounts. The time delay between administration of the components will vary depending on the exact properties of the components, their interaction, and their respective half-lives.

[0037] As used herein, "separately" means that the administration of one formulation / composition (e.g., pharmaceutical composition) is spaced sufficiently apart from the administration of the other formulation / composition that, when a second formulation is administered, the formulation administered initially is no longer present in the bloodstream in therapeutically effective amounts.

[0038] The pharmaceutical composition of the present invention can be used for the prevention or treatment of cancer. In the present invention, the term "cancer" refers to cancer that is caused by a mass generated by abnormal cell growth in various solid organs such as the bladder, breast, intestine, kidney, lung, liver, brain, esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate and skin.

[0039] Cancers that can be prevented or treated by the pharmaceutical composition of the present invention include breast cancer, ovarian cancer, endometrial cancer, small cell lung cancer, neuroendocrine tumors, biliary tract cancer, bile duct carcinoma, urothelial cancer, bladder cancer, pancreatic cancer, prostate cancer, gastric cancer, esophageal cancer, colon cancer, thyroid cancer, blood cancer, and among the above-mentioned cancers, homologous recombination deficient cancers, and preferably gastric cancer.

[0040] In the present invention, the term "gastric cancer" refers to cancer occurring in the stomach, and includes other diseases such as adenocarcinoma occurring in the glandular cells of the gastric wall mucosa, lymphomas occurring in the lymphatic system, and gastrointestinal stromal tumors occurring in interstitial tissue.

[0041] In the present invention, the term "prevention" means any action of suppressing or delaying the occurrence, spread, and recurrence of cancer by administering the pharmaceutical composition of the present invention, and the term "treatment" means any action of improving or beneficially changing the symptoms of a subject suspected of or having cancer by administering the pharmaceutical composition of the present invention.

[0042] In the present invention, the term "administration" means providing a predetermined substance to a patient by any suitable method, and the administration route may be any common route as long as the pharmaceutical composition of the present invention can achieve target regulation. The administration method may include, but is not limited to, oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration, or intrarectal administration. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of transporting an active ingredient to a target cell.

[0043] The pharmaceutical composition of the present invention may be administered orally and may be formulated into a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, etc. by a conventional method. The pharmaceutical composition of the present invention may also be used in the form of an external preparation, suppository, or external skin preparation (ointment, lotion, spray, patch, cream, powder, suspension, gel, gel form, etc.).

[0044] In some embodiments, the first pharmaceutical composition is administered once a day. In some embodiments, the first pharmaceutical composition is an oral dosage form. In some embodiments, the first pharmaceutical composition is a tablet. In some embodiments, the first pharmaceutical composition may be administered in a dose of about 1 mg to about 50 mg. In some embodiments, the first pharmaceutical composition may be administered in a dose of about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, or about 50 mg per administration. In some embodiments, the first pharmaceutical composition may be administered in a dose of about 10 mg, about 20 mg, about 40 mg, about 80 mg, about 120 mg, or about 160 mg per administration.

[0045] Preferred parenteral administration methods and formulations of the pharmaceutical composition of the present invention include intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, drip injection, etc. Injections can be prepared using aqueous solvents such as physiological saline and Ringer's solution, and non-aqueous solvents such as vegetable oils, high fatty acid esters (e.g., ethyl oleate, etc.), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.). The injections may also contain pharmaceutical carriers such as stabilizers for preventing deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for adjusting pH, and preservatives for inhibiting the growth of microorganisms (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

[0046] In some embodiments, the second pharmaceutical composition is in an injection dosage form. In some embodiments, the second pharmaceutical composition is administered once daily. In some embodiments, the second pharmaceutical composition is administered intravenously. In some embodiments, the second pharmaceutical composition is administered continuously.

[0047] The pharmaceutical composition of the present invention may further comprise a pharma- ceutically acceptable carrier. In the present invention, "pharma-ceutically acceptable" means that the composition exhibits non-toxic properties to cells or humans exposed to the composition. As the carrier, any substance known in the art, such as a buffer, a preservative, an analgesic, a solubilizer, an isotonicity agent, a stabilizer, a base, an excipient, or a lubricant, may be used without limitation.

[0048] In the present invention, the term "excipient" refers to a pharma- ceutically acceptable inactive ingredient used in the preparation of a pharmaceutical formulation. The excipient may include, but is not limited to, a diluent, a binder, a disintegrant, a super-strong disintegrant, a flow agent, a pH regulator, a lubricant, a filler, a stabilizer, an antioxidant, a film coating agent, etc. The excipient used in the present invention may be appropriately selected by referring to literature known in the art, for example, [Handbook of Pharmaceutical Excipients, Sixth edition, Pharmaceutical Press], which is incorporated herein by reference.

[0049] The carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. When formulated, it may be formulated using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

[0050] Examples of solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. Such solid preparations can be prepared by mixing the compound represented by Chemical Formula 1 with one or more excipients such as starch, calcium carbonate, sucrose or lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate can also be used. Examples of liquid preparations for oral administration include suspensions, oral liquids, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, flavorings, and preservatives can also be included.

[0051] Preparations for parenteral administration include sterile solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried agents, suppositories, etc. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used. As bases for suppositories, Witepsol (registered trademark), macrogol, Tween 61, cacao butter, laurin, glycerogelatin, etc. may be used.

[0052] The pharma- ceutical effective amount of the pharmaceutical composition of the present invention is such that the pharmaceutical composition of the present invention can be used alone in mammals, including humans, or can be used in combination with other active ingredients, kits, products, or combination preparations contained in the pharmaceutical composition to prevent, eliminate, or reduce the harmful effects of cancer.

[0053] In the present invention, a "pharmaceutical effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to any medical treatment, and the level of a "therapeutically effective amount" may be determined by including the patient's health condition, the type and severity of the cancer, drug activity, drug sensitivity, administration method, administration time, administration route and excretion rate, treatment duration, co- or simultaneous drugs, and other factors well known in the medical field. It is understood that the dose administered in the present invention is also lower for each compound in the composition than the "therapeutically effective amount" defined for each compound used alone or in combination with other treatments other than the above-mentioned combination therapy. In particular, the pharmaceutical composition, kit, product, and combination of the present invention may reduce tumor progression, reduce tumor burden, induce tumor regression, and / or prevent metastasis or the development of cancer in a mammalian host. In cancer treatment, the pharmaceutical composition of the present invention may be administered in a therapeutically effective amount.

[0054] In one embodiment, the daily dose of the compound represented by formula 1 of the present invention is about 1 mg to about 400 mg, preferably about 2 mg to about 240 mg, more preferably about 5 mg to about 160 mg, and even more preferably about 5 mg to about 120 mg. In addition, the daily dose of irinotecan, an anticancer agent of the present invention, is about 10 mg / m 2 to about 350 mg / m 2 , preferably about 25 mg / m 2 to about 100 mg / m 2 , more preferably 25 mg / m 2 to about 75 mg / m 2 In some embodiments, the anticancer drug (e.g., irinotecan) is administered at a dose of about 30 mg / m 2 to about 120 mg / m 2 In some embodiments, the anticancer drug (e.g., irinotecan) may be administered intravenously at a dose of about 50 mg / m 2 , about 75mg / m 2 or about 100 mg / m 2 In some embodiments, the anticancer drug (e.g., irinotecan) may be administered at a dose of about 125 mg / m2 , 180 mg / m 2 or about 350 mg / m 2 In some embodiments, the second pharmaceutical composition is administered intravenously at a dose of about 30 mg / m 2 to about 120 mg / m 2 In some embodiments, the second pharmaceutical composition may be administered intravenously at a dose of about 50 mg / m 2 , about 75mg / m 2 or about 100 mg / m 2 However, due to the characteristics of cancer, which is the target disease of the pharmaceutical composition of the present invention, the dosage of each component may vary greatly depending on the above conditions, and is not limited to the above examples.

[0055] As used herein, the term "about" or "approximately" refers to a ±10% variation from the cited quantitative value (and includes the cited quantitative value itself), unless otherwise expressly stated or inferred by the context. For example, about 100 mg / m 2 The dose was 100±10% mg / m 2 90 mg / m 2 to 110 mg / m 2 Including up to.

[0056] In the pharmaceutical composition for combined administration of the present invention, the appropriate administration cycle of (i) the compound represented by Chemical Formula 1 or a pharma- ceutical acceptable salt thereof, and (ii) the anticancer agent may be determined according to the dose. In the present invention, (i) the compound represented by Chemical Formula 1 or a pharma- ceutical acceptable salt thereof may be administered once a day to once a week, preferably once a day, and (ii) the anticancer agent may be administered once a day to once a week, preferably once a week.

[0057] In one embodiment of the present invention, the administration cycle of the pharmaceutical composition for combined administration of the present invention is, but is not limited to, 14 to 21 days. In one embodiment of the present invention, when the administration cycle is 14 days, (i) from day 1 to day 7, day 1 to day 5, or day 2 to day 6, the compound represented by Formula 1 or a pharma- ceutical acceptable salt may be orally administered or injected once a day, and (ii) from day 1 or day 2 to day 6, the anticancer agent may be injected once a day.

[0058] In one embodiment of the present invention, when the (1) cycle of administration is 21 days, (i) from day 1 to day 7, from day 1 to day 5, or from day 2 to day 6, the compound represented by Formula 1, or a pharma- ceutically acceptable salt, may be administered orally or by injection once a day, and (ii) from day 1, or from day 2 to day 6, the anticancer agent may be administered by injection once a day.

[0059] In some embodiments, the first pharmaceutical composition is administered in a cycle, where the cycle comprises administering the first pharmaceutical composition for about 14 days or about 21 days. For example, in some embodiments, the first pharmaceutical composition is administered to a subject in need thereof for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, or 14 days.

[0060] In some embodiments, the second pharmaceutical composition is administered in a cycle, where the cycle comprises administering the second pharmaceutical composition for about 14 days or about 21 days. In some embodiments, the second pharmaceutical composition should be administered in a cycle, where the cycle is repeated at least once. For example, in some embodiments, the second pharmaceutical composition is administered to a subject in need thereof for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days or 14 days.

[0061] In some embodiments, in one cycle, a first pharmaceutical composition is administered on days 1 through 7, days 1 through 5, or days 2 through 6, and a second pharmaceutical composition is administered on days 1, or days 2 through 6, where the first pharmaceutical composition is administered once daily and the second pharmaceutical composition is administered once daily, and a cycle proceeds for 14 or 21 days. In some embodiments, in one cycle, a first pharmaceutical composition comprising 40 mg, 20 mg, or 10 mg of a compound of Formula I, or a pharma- ceutical acceptable salt thereof, is administered once daily, and a second pharmaceutical composition is administered at 100 mg / m 2 is administered at an intravenous dose of

[0062] The pharmaceutical composition for combined administration of the present invention may be administered as an individual therapeutic agent, may be administered again in combination with other therapeutic agents, may be administered sequentially or simultaneously with existing therapeutic agents, may be administered once or several times. It is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects, taking into consideration all the above factors.

[0063] In the present invention, the term "kit", "product" or "concomitant agent" specifically defines a "component kit" in the sense that the compound represented by Formula 1 as defined above is administered alone or in combination with a fixed amount of a different combination agent from the combination anticancer agent, i.e., administered simultaneously or at different times. The components of the component kit can then be administered, for example, simultaneously or sequentially, i.e., at different times. That is, any components of the component kit can be administered simultaneously or at different time intervals. In the combination formulation, the ratio of the total dosage of the compound represented by Formula 1 and the combination anticancer agent can also be varied. The combination anticancer agents can be administered by the same or different routes. In some embodiments, the first pharmaceutical composition is present in a single dosage form and the second pharmaceutical composition is present in a separate dosage form. In some embodiments, the first pharmaceutical composition is administered simultaneously, separately or sequentially.

[0064] In the present invention, the term "treatment" refers to curative, symptomatic, and preventive treatment. The pharmaceutical compositions, kits, products, and combination agents of the present invention can be used in humans with cancer or tumors at early or late stages of cancer progression.

[0065] The pharmaceutical compositions, kits, products, and combinations of the invention do not necessarily cure a cancer patient, but may improve the patient's condition by delaying, slowing, or preventing further progression of the disease.

[0066] Throughout this specification, when the term "cancer treatment" is mentioned in connection with the pharmaceutical composition of the present invention, it means (a) a method of treating cancer, which comprises administering the pharmaceutical composition of the present invention to a subject in need of such treatment, (b) the use of the pharmaceutical composition of the present invention for the treatment of cancer, (c) the use of the pharmaceutical composition of the present invention for the manufacture of a medicament for treating cancer, and / or (d) the pharmaceutical composition of the present invention for use in the treatment of cancer.

[0067] Yet another aspect of the present invention provides a method for preventing or treating cancer, comprising administering to an individual in need thereof a pharmaceutical composition comprising (i) a compound represented by Formula 1, or a pharma- ceutical acceptable salt thereof, and (ii) at least one anti-cancer agent.

[0068] In one embodiment, contemplated herein is a method of preventing or treating cancer in a subject in need thereof, the method comprising administering a pharmaceutical composition described herein.

[0069] In one aspect, the present invention provides a method for administering to a subject: (i) a first pharmaceutical composition comprising a compound represented by Formula 1, or a pharma- ceutically acceptable salt thereof; [ka] and (ii) A method of treating cancer in a subject in need thereof comprising administering a second pharmaceutical composition comprising an anti-cancer agent. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is advanced gastric cancer. In some embodiments, the anti-cancer agent is selected from irinotecan and oxaliplatin. In some embodiments, the anti-cancer agent is irinotecan. In some embodiments, the anti-cancer agent is oxaliplatin. In some embodiments, the pharmaceutically acceptable salt is a hydrochloride salt.

[0070] Yet another aspect of the present invention provides the use of a compound of formula 1, or a pharma- ceutically acceptable salt thereof, for the manufacture of a first medicament, and the use of an anti-cancer agent for the manufacture of a second medicament, wherein the first medicament and the second medicament are for preventing or treating cancer in a subject in need thereof. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is advanced gastric cancer. In some embodiments, the anti-cancer agent is selected from irinotecan and oxaliplatin. In some embodiments, the anti-cancer agent is irinotecan. In some embodiments, the anti-cancer agent is oxaliplatin. In some embodiments, the pharma- ceutically acceptable salt is a hydrochloride salt.

[0071] Yet another aspect of the present invention provides the use of a first pharmaceutical composition comprising a compound of formula 1, or a pharma- ceutically acceptable salt thereof, for the manufacture of a first medicament, and a second pharmaceutical composition comprising an anti-cancer agent, for the manufacture of a second medicament, wherein the first medicament and the second medicament are for preventing or treating cancer in a subject in need thereof. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is advanced gastric cancer. In some embodiments, the anti-cancer agent is selected from irinotecan and oxaliplatin. In some embodiments, the anti-cancer agent is irinotecan. In some embodiments, the anti-cancer agent is oxaliplatin. In some embodiments, the pharma- ceutically acceptable salt is a hydrochloride salt.

[0072] In some embodiments, the anti-cancer drug is selected from the group consisting of chemotherapy drugs, targeted therapy drugs, immunotherapy drugs, and hormone therapy drugs. In some embodiments, the chemotherapy drug is selected from the group consisting of alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, and mitotic inhibitors. In some embodiments, the targeted therapy drug is selected from the group consisting of angiogenesis inhibitors, monoclonal antibodies, proteasome inhibitors, and signal transduction inhibitors. In some embodiments, the immunotherapy drug is selected from the group consisting of checkpoint inhibitors, chimeric antigen receptor (CAR) T cell therapy, and cytokines. In some embodiments, the hormone therapy drug is selected from the group consisting of aromatase inhibitors, selective estrogen receptor modulators, estrogen receptor antagonists, luteinizing hormone releasing hormone enhancers, antiandrogens, and CYP17 inhibitors. In some embodiments, the anti-cancer drug is selected from irinotecan and oxaliplatin. In some embodiments, the anti-cancer drug is irinotecan. In some embodiments, the anti-cancer agent is oxaliplatin. In some embodiments, the pharma- ceutically acceptable salt is a hydrochloride salt.

[0073] In some embodiments, the method or use comprises administering a first pharmaceutical composition and a second pharmaceutical composition in a mixture, hi some embodiments, the method or use comprises administering the first pharmaceutical composition and the second pharmaceutical composition in separate formulations, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously or sequentially.

[0074] In some embodiments, the method or use includes administering the first pharmaceutical composition and the second pharmaceutical composition in separate dosage forms. In some embodiments, the method or use includes administering the first pharmaceutical composition in an oral dosage form and the second pharmaceutical composition in an injectable dosage form. In some embodiments, the method or use includes administering the first pharmaceutical composition in a tablet and administering the second pharmaceutical composition intravenously.

[0075] In some embodiments, the method or use comprises administering the first pharmaceutical composition and the second pharmaceutical composition once a day, respectively. In some embodiments, the method or use comprises administering the first pharmaceutical composition at about 1 mg to about 180 mg. In some embodiments, the method or use comprises administering the first pharmaceutical composition at about 5 mg to about 170 mg. In some embodiments, the method or use comprises administering the first pharmaceutical composition at about 10 mg to about 160 mg. In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 10 mg, about 20 mg, about 40 mg, about 80 mg, about 120 mg, or about 160 mg per administration. In some embodiments, the first pharmaceutical composition comprises about 1 mg to about 50 mg of the compound of formula I. In some embodiments, the first pharmaceutical composition comprises about 5 mg to about 40 mg of a compound represented by Formula I. In some embodiments, the first pharmaceutical composition comprises about 5 mg, about 10 mg, about 20 mg, or about 40 mg of a compound represented by Formula I. In some embodiments, the first pharmaceutical composition is administered to a subject in need thereof with at least 8 ounces of water.

[0076] In some embodiments, the method or use comprises administering the second pharmaceutical composition at a dose of about 30 mg / m 2 to about 120 mg / m 2 In some embodiments, the method or use includes administering the second pharmaceutical composition intravenously at a dose of about 50 mg / m 2 , about 75mg / m 2 or about 100 mg / m 2In some embodiments, the method or use includes administering the first pharmaceutical composition and the second pharmaceutical composition in a cycle, wherein a cycle has a duration of about 14 days or 21 days. In some embodiments, the cycle is repeated at least once. In some embodiments, the second pharmaceutical composition includes administering an anti-cancer agent (e.g., irinotecan) at a concentration of about 20 mg / mL. In some embodiments, the second pharmaceutical composition includes 5% dextrose or 0.9% sodium chloride. In some embodiments, the anti-cancer agent (e.g., irinotecan) is administered for about 90 minutes. In some embodiments, the anti-cancer agent (e.g., irinotecan) is administered for more than about 90 minutes.

[0077] In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 40 mg and the second pharmaceutical composition at a dose of about 50 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 40 mg and the second pharmaceutical composition at a dose of about 75 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 10 mg and the second pharmaceutical composition at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 20 mg and the second pharmaceutical composition at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 40 mg and the second pharmaceutical composition at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 80 mg and the second pharmaceutical composition at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition at a dose of about 120 mg and the second pharmaceutical composition at a dose of about 100 mg / m 2The method includes administering the compound intravenously at a dose of

[0078] In some embodiments, the methods or uses include administering a first pharmaceutical composition at a dose of about 40 mg once daily and a second pharmaceutical composition at a dose of about 50 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition intravenously at a dose of about 40 mg once daily and the second pharmaceutical composition intravenously at a dose of about 75 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition intravenously at a dose of about 10 mg once daily and the second pharmaceutical composition intravenously at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition intravenously at a dose of about 20 mg once daily and the second pharmaceutical composition intravenously at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition intravenously at a dose of about 40 mg once daily and the second pharmaceutical composition intravenously at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition intravenously at a dose of about 80 mg once daily and the second pharmaceutical composition intravenously at a dose of about 100 mg / m 2 In some embodiments, the method or use comprises administering the first pharmaceutical composition intravenously at a dose of about 120 mg once daily and the second pharmaceutical composition intravenously at a dose of about 100 mg / m 2 The method includes administering the compound intravenously at a dose of

[0079] In some embodiments, the present invention provides a method for administering to a subject in need of cancer treatment a first pharmaceutical composition comprising: (i) a compound represented by Formula 1, or a pharma- ceutically acceptable salt thereof; [ka] and (ii) A method of treating cancer comprising administering a second pharmaceutical composition comprising irinotecan. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is advanced gastric cancer. In some embodiments, the present invention provides a method for administering to a subject in need of cancer treatment a first pharmaceutical composition comprising: (i) a compound represented by Formula 1, or a pharma- ceutically acceptable salt thereof; [ka] and (ii) a second pharmaceutical composition comprising oxaliplatin is administered. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is advanced gastric cancer. In the present invention, the term "subject" refers to any animal, including humans, in which cancer has occurred or may occur, and cancer can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to a subject. The pharmaceutical composition of the present invention can be administered in combination with an existing therapeutic agent.

[0080] The pharmaceutical compositions of the present invention may be used in combination with other known therapies for the treatment of diseases, including, but not limited to, chemotherapy, radiation therapy, hormone therapy, bone marrow transplantation, stem cell replacement therapy, other biological therapies, immunotherapy, and the like.

[0081] As described herein, the therapeutic efficacy of the combination of pharmaceutical compositions of the present invention may be determined by a variety of endpoints. In some embodiments, the therapeutic efficacy of a treatment is determined by assessing improvement according to the response evaluation criteria in solid tumors (RECIST), which assesses objective response rate (ORR), progression-free survival (PFS), disease control rate (DCR), duration of response (DOR), time to response (TTR), best overall response (BOR), time to progression (TTP), or overall survival (OS).

[0082] Furthermore, the therapeutic efficacy of the combination of the pharmaceutical composition of the present invention can be determined by the reduction of tumor volume.For example, the therapeutic efficacy of the combination can be determined by the reduction of tumor volume growth after administration of the combination, compared with administration of the anticancer drug alone.The therapeutic efficacy of the combination can also be determined by the inhibitory effect on tumor cell proliferation.For example, the therapeutic efficacy of the combination can be determined through the inhibitory effect on tumor cell proliferation after administration of the combination, compared with administration of the anticancer drug alone. Effect of the Invention

[0083] It has been confirmed that the compound represented by Chemical Formula 1 of the present invention or a pharma- ceutically acceptable salt thereof, when administered in combination with a known anticancer drug, exhibits a synergistic complementary effect and exhibits excellent anticancer activity, compared to administration of the anticancer drug alone. When the compound represented by Chemical Formula 1 of the present invention or a pharma- ceutically acceptable salt thereof is administered in combination with an anticancer drug, specifically irinotecan or oxaliplatin, it is possible to obtain the same or better anticancer effect at a lower dose than that of the known anticancer drug, and to obtain an excellent effect on cancers limited by existing treatment methods. It is also possible to obtain an effect of reducing dose-dependent side effects. The pharmaceutical composition for combined administration of the present invention is particularly useful as an anticancer drug, since it exhibits a remarkable synergistic effect in the prevention or treatment of cancers, including solid tumors such as gastric cancer, by combining the compound represented by Chemical Formula 1 or a pharma- ceutically acceptable salt thereof (first active ingredient) with an anticancer drug (second active ingredient). [Brief description of the drawings]

[0084] [Figure 1] 1 is a graph showing a comparison of cell viability in a case where the hydrochloride of the compound represented by Chemical Formula 1 is administered in combination with irinotecan and in a case where irinotecan is administered alone. [Diagram 2] 1 is a graph showing a comparison of cell viability in a case where the hydrochloride of the compound represented by Chemical Formula 1 is administered in combination with oxaliplatin and in a case where oxaliplatin is administered alone. [Diagram 3] 1 is a graph showing the effect on tumor volume (mean±SEM) of administering the hydrochloride of the compound of Formula 1 in combination with irinotecan versus administering the hydrochloride of the compound of Formula 1 alone in a subcutaneous MKN45 human gastric cancer xenograft model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0085] The present invention will be described in detail below with reference to examples. However, the following examples are for illustrating the present invention, and the present invention is not limited thereto.

[0086] In order to evaluate the preventive and therapeutic effects of the pharmaceutical composition of the present invention on the prevention and treatment of cancer, the following experiments were carried out.

[0087] Experimental Example 1 1.1. Preparation and handling of DMSO stock solutions The hydrochloride salt of the compound represented by Chemical Formula 1 (MW 442.92 (hereinafter referred to as "the hydrochloride salt compound of Chemical Formula 1")) was purchased from Ildong Pharmaceutical. Irinotecan hydrochloride (MW 623.14) was purchased from Sigma Aldrich.

[0088] Each of the drugs was dissolved in 100% DMSO to prepare a 10 mM stock solution, which was then divided into single doses and stored at -20°C.

[0089] A 10 mM stock solution containing the hydrochloride compound of Chemical Formula 1 and irinotecan was serially diluted in DMSO at a ratio of 1 / 10 to prepare stock solutions for each treatment concentration. Here, DMSO was used so that the final content did not exceed 1.1% to treat the cells.

[0090] Five human gastric cancer cell lines (i.e., AGC, SNU-668, SNU-216, SNU-484, and SNU-638) were used as cell lines. The source of the cell lines was the Korea Cell Line Bank (KCLB). The culture medium used was a complete growth medium composed of 90% RPMI1640 and 10% heat-inactivated fetal bovine serum.

[0091] 1.2. Thawing and subculturing of frozen cells The frozen cells were thawed in a 37°C water bath, suspended in culture medium, and centrifuged at 1,200 rpm for 2 minutes. The precipitated cells were suspended in culture medium, sprayed onto a culture dish, and cultured at 37°C and 5% CO2. The cells were cultured periodically at intervals of 3 to 5 days, reflecting the growth rate of each cell.

[0092] 1.3. Cell Dispensing and Drug Treatment To remove cells from the culture dish, the cells were treated with trypsin and then reacted in a 5% CO2, 37°C incubator. After the reaction was completed, the collected cells were suspended in culture medium and centrifuged at 1,200 rpm for 3 minutes. The cell suspension was placed in a hemocytometer and the number of cells per unit volume was calculated. According to the conditions set for each cell line, the cell suspension was plated into a 96-well plate and then stabilized at 5% CO2, 37°C for 24 hours.

[0093] A 10 mM stock solution containing irinotecan was serially diluted in DMSO at a 1 / 10 ratio to prepare 100X stock solutions for each treatment concentration, and a 1,000X stock solution containing 1 mM hydrochloride compound of Formula 1 was prepared. A stock solution with a final treatment concentration of 1 μM was prepared. To minimize the effect of the solvent on the cells, each well contained 1.1% DMSO. The stock solutions of each concentration were added to the culture medium to give a 10-fold (10X) concentration, and the cells were then treated with a 1-fold (1X) concentration. Each cell line was treated with irinotecan alone or with the hydrochloride compound of Formula 1 in combination with irinotecan for 5 days.

[0094] 1.4.MTT analysis After each drug treatment, 100 μL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide solution (Sigma Aldrich) was added to each well, and the plate was incubated at 37° C. for 2 hours, after which the culture medium was removed. After dissolving the formazan crystals in 100 μL of DMSO, the absorbance of each cell was measured at 570 nm using a Synergy H4 microplate reader (BioTek).

[0095] Data Analysis The absorbance was calculated as a percentage of the concentration of the drug compared to the negative control group (100%) containing only the solvent (DMSO). IC 50 was calculated by nonlinear regression analysis of three or four parameter fits (GraphPad Prism version 7.04).

[0096] Using MTT analysis, the sensitivity of each cell line to irinotecan with or without the addition of 1 μM of the hydrochloride compound of Chemical Formula 1 was confirmed through the viability, and the results are shown in Figure 1. As shown in Figure 1, the administration of irinotecan alone showed a cell killing effect, while the cell killing rate was even higher when irinotecan was administered in combination with the compound of Chemical Formula 1. Therefore, it was confirmed that the cancer growth inhibitory effect obtained by such combination administration increased in a dose-dependent manner. The calculated IC 50 The values ​​are shown in Table 1. [Table 1]

[0097] Referring to Table 1, the IC of the five cell lines in Example 1 50 It can be seen that the values ​​were 1.3 to 3.1 times lower than those in Comparative Example 1. Therefore, in the experiment using the gastric cancer cell line, it was confirmed that the pharmaceutical composition of the present invention exhibits a superior tumor cell proliferation inhibitory effect compared to the administration of an anticancer agent alone.

[0098] Experimental Example 2 2.1. Preparation and handling of DMSO stock solutions The hydrochloride salt of the compound represented by Chemical Formula 1 (MW 442.92 (hereinafter referred to as "the hydrochloride salt of Chemical Formula 1")) was purchased from Ildong Pharmaceutical. Oxaliplatin (MW 397.29) was purchased from Sigma Aldrich.

[0099] Each of the drugs was dissolved in 100% DMSO to prepare a 10 mM stock solution, which was then divided into single doses and stored at -20°C.

[0100] A 10 mM stock solution containing the hydrochloride compound of Formula 1 and oxaliplatin was serially diluted with DMSO at a ratio of 1 / 10 to prepare stock solutions for each treatment concentration, where the final content of DMSO was not more than 1.1% to treat the cells.

[0101] Five human gastric cancer cell lines (i.e., AGC, SNU-668, SNU-216, SNU-484, and SNU-638) were used as cell lines. The source of the cell lines was KCLB. The medium used was a complete growth medium composed of 90% RPMI1640 and 10% heat-inactivated fetal bovine serum.

[0102] 2.2. Thawing and subculturing of frozen cells The frozen cells were thawed in a 37°C water bath, suspended in culture medium, and centrifuged at 1,200 rpm for 2 minutes. The precipitated cells were suspended in culture medium, sprayed onto a culture dish, and cultured at 37°C and 5% CO2. The cells were cultured periodically at intervals of 3 to 5 days to reflect the growth rate of each cell.

[0103] 2.3. Cell Dispensing and Drug Treatment To remove cells from the culture dish, the cells were treated with trypsin and then incubated in a 5% CO2, 37°C incubator. After the reaction was completed, the collected cells were suspended in culture medium and centrifuged at 1,200 rpm for 3 minutes. The cell suspension was placed in a hemocytometer and the number of cells per unit volume was calculated. According to the conditions set for each cell line, the cell suspension was dispensed into a 96-well plate and then stabilized at 5% CO2, 37°C for 24 hours.

[0104] A 10 mM stock solution of oxaliplatin was serially diluted in DMSO at a 1 / 10 ratio to prepare 100-fold (100X) stock solutions for each treatment concentration, and a 1,000-fold (1000X) stock solution containing 1 mM Formula 1 hydrochloride was prepared to prepare a stock solution with a final treatment concentration of 1 μM. To minimize the effect of the solvent on the cells, each well contained 1.1% DMSO. The stock solutions of each concentration were added to the culture medium to give a 10-fold (10X) concentration, and the cells were then treated with 1X. Each cell line was treated with oxaliplatin alone or with the hydrochloride compound of Formula 1 in combination with oxaliplatin for 5 days.

[0105] 2.4.MTT analysis After each drug treatment, 100 μL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide solution (Sigma Aldrich) was added to each well, the plate was incubated at 37° C. for 2 hours, and the culture medium was removed. After dissolving the formazan crystals in 100 μL of DMSO, the absorbance of each cell was measured at 570 nm using a Synergy H4 microplate reader (BioTek).

[0106] The absorbance was compared to the negative control group (100%) containing only the solvent (DMSO), and the activity of each drug at each concentration was expressed as a percentage and calculated. IC 50 was calculated by nonlinear regression analysis with three or four parameter fits (GraphPad Prism 7.04).

[0107] Using MTT analysis, the sensitivity of each cell line was confirmed through the viability of oxaliplatin with or without the addition of 1 μM of the hydrochloride compound of Chemical Formula 1, and the results are shown in Figure 2. As shown in Figure 2, the administration of oxaliplatin alone showed a cell killing effect, while the cell killing rate was even higher when oxaliplatin was administered in combination with the compound of Chemical Formula 1. Therefore, it was confirmed that the cancer growth inhibitory effect obtained by such combination administration increased in a dose-dependent manner. The calculated IC 50 The values ​​are shown in Table 2. [Table 2]

[0108] With reference to Table 2, IC of specific cell lines in Example 2 50 It can be seen that the values ​​were 2.7 to 6.8 times lower than those in Comparative Example 2. Therefore, in the experiment using the gastric cancer cell line, it was confirmed that the pharmaceutical composition of the present invention exhibits a superior tumor cell proliferation inhibitory effect compared to the administration of an anticancer agent alone.

[0109] Experimental Example 3 3.1.Cell culture MKN45 tumor cells were grown in RPMI1640 supplemented with 10% fetal bovine serum at 37°C and 5% CO2. Cells were harvested before tumor implantation and then counted using a cell counter.

[0110] Tumor implantation To perform tumor implantation, PBS tumor suspension (MKN45 tumor cells 3 × 10 6 Each mouse was inoculated subcutaneously with 0.1 ml of the vaccine in the right upper flank.

[0111] 3.3. Randomization Randomization occurred when mice had a mean tumor size of approximately 150 mm 3A total of 40 mice were used in the study, with 10 mice per group, and they were assigned to 4 groups as shown in Table 3 (Example 3 and Comparative Examples 3 to 5). The randomization was performed using the "matched distribution" method (Study Director, TM The analysis was performed based on the software (version 3.1.399.19) where the grouping date was displayed as 0. [Table 3]

[0112] 3.4. Observation and collection After tumor inoculation, animals were checked daily for morbidity and mortality. During routine monitoring, animals were monitored for the effects of tumor growth and treatment on behaviors such as mobility, food and water intake, weight gain / loss (weighted twice a week after randomization), eye / hair lubrication, and other abnormalities. Mortality and observed clinical symptoms were recorded in detail for each animal.

[0113] After randomization, tumor volumes were measured twice weekly using calipers and calculated according to the following formula: V = (length x width x width) / 2 V(mm 3 ) is the tumor volume, L is the tumor length (the longest dimension of the tumor), and W is the tumor width (the widest width of the tumor perpendicular to L). In addition to tumor and body weight measurements, administration was performed in a Laminar Flow Cabinet.

[0114] Body weight and tumor volume were determined by the Study Director. TM The weight loss was measured using the software (version 3.1.399.19). As a result, no significant weight loss was observed in any group.

[0115] 3.5. Drug Formulations for Coadministration The drug formulations for co-administration are as shown in Table 4. [Table 4]

[0116] 3.6. Tumor volume and growth curve The mean tumor growth curves at various time points are illustrated in FIG. [Table 5]

[0117] Referring to Table 5, it can be seen that the tumor volume growth in Example 3 is smaller than that in Comparative Examples 4 and 5. That is, in the experiment using mice, it was confirmed that the pharmaceutical composition for combined administration of the present invention significantly reduces the tumor volume growth compared to the administration of an anticancer drug alone.

[0118] Tumor growth inhibition In a subcutaneous MKN45 human gastric cancer xenograft model, tumor growth inhibition by a single preparation of the hydrochloride compound of Formula 1 and a combination preparation of the hydrochloride compound of Formula 1 and irinotecan was measured, and the results are shown in Table 6. [Table 6]

[0119] Referring to Table 6, when the hydrochloride compound of Chemical Formula 1 in Example 3 (2 mg / kg (QD)) and irinotecan (5 mg / kg (QW)) were used in combination in the MKN45 model, the tumor size on day 21 was 487.84 mm 3 The tumor growth inhibition (TGI) value on the 21st day was 46.13%. In Comparative Example 3 using the vehicle control group, the average tumor size on the 21st day was 905.67 mm 3The TGI value on day 21 of Comparative Example 4, in which 2 mg / kg of the hydrochloride compound of formula 1 was administered QD (quaque die), was 19.69%, and the TGI value on day 21 of Comparative Example 5, in which 5 mg / kg of irinotecan was administered QW, was 28.91%. Here, the TGI is calculated by TGI%=(1-ΔT / ΔC)×100 (where T and C are the mean tumor volumes of the treatment and control groups, respectively). If the in vivo TGI is 70% or more, it can be considered sensitive to the treatment, if the in vivo TGI is 40-70%, it can be considered partially responsive to progressive disease, and if the in vivo TGI is 40% or less, it can be considered non-responsive to the treatment (Cancer Res 2006; 66: (14). July 15, 2006, Chronic Administration of Valproic Acid Inhibits Prostate Cancer Cell Growth In vitro and In vivo).

[0120] In other words, the combination of the hydrochloride compound of Chemical Formula 1 and irinotecan of the present invention can be seen to exhibit significant antitumor efficacy compared to the vehicle control group, a single formulation of the hydrochloride compound of Chemical Formula 1, and a single formulation of irinotecan.

[0121] Experimental Example 4 Purpose of the experiment The primary objective of this study is to evaluate the safety, maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D) of the combination pharmaceutical composition of the present invention in patients with advanced gastric cancer.

[0122] The secondary objectives of this study were to evaluate the efficacy of the combination of the pharmaceutical compositions of the present invention as defined by objective response rate (ORR), progression-free survival (PFS), disease control rate (DCR), duration of response (DOR), time to response (TTR), best overall response (BOR), time to progression (TTP), and overall survival (OS) as assessed by Response Evaluation Criteria in Solid Tumors (RECIST) v1.1, to evaluate the pharmacokinetics (PK) of a single ascending dose (SAD) of the compound of formula 1 when administered together with irinotecan in patients with advanced gastric cancer, and to evaluate population PK modeling of the compound of formula 1 and irinotecan when administered in combination with the compound of formula 1 and irinotecan in patients with advanced gastric cancer.

[0123] End point of the experiment The primary endpoints of the study are to determine the MTD and RP2D of the pharmaceutical compounds of the invention, to determine the incidence of DLTs and adverse events (AEs) / serious adverse events (SAEs).

[0124] Secondary endpoints of the study were related to safety and efficacy, where safety endpoints represented changes from baseline in stability parameters including physical examination, vital signs and clinical laboratory results including electrocardiogram (ECD), hematology, coagulation studies, clinical chemistry, and urinalysis, and efficacy evaluation variables represented the number of patients with ORR, PFS, BOR, TTP, DCR, DOR, TTR, and OS as assessed by RECIST v1.1.

[0125] An exploratory endpoint of the study is to evaluate the correlation between efficacy of the compound of Formula 1 and ATM levels (high and low) in patients with advanced gastric cancer.

[0126] Experimental design Each part of the study was divided into three periods: screening, treatment, and follow-up (including survival follow-up).

[0127] Here, the dose determination includes the group "compound of formula 1 + irinotecan." [Table 7]

[0128] For the dose expansion, approximately 45 patients with advanced gastric cancer (15 per group) who met all eligibility criteria were enrolled in the open dose expansion portion to additionally explore the safety, tolerability, and efficacy of the compound of formula 1 in combination with irinotecan.

[0129] Patients were administered an appropriate dose (potential RP2D) of the compound of formula 1 QD in combination with irinotecan in 14-day cycles until disease progression, unacceptable toxicity, or other withdrawal criteria were met. Treatment cycles were administered consecutively without interruption, except as needed to manage adverse events (AEs). Treatment with the compound of formula 1 may be continued after disease progression has been identified if the investigator determines that continued treatment would be of clinical benefit to the patient.

[0130] Selection of experimental population (patient selection) Our experimental population includes patients with advanced gastric cancer, defined as metastatic or recurrent / refractory progressive disease in the third or fourth line. Patients in the dose determination portion must have evaluable disease. Patients in the dose expansion portion must have measurable disease at baseline by imaging (computed tomography (CT) or magnetic resonance imaging (MRI)).

[0131] The patient selection criteria were as follows: ● Patients who have signed an Independent Ethics Committee (IEC) / Institutional Review Board (IRB) approved informed consent form (ICF) for screening tests prior to carrying out clinical trial procedures. • Males or females who are at least 18 years of age at the time of consent (i.e., screening), or if the legal age for consenting to participate in a clinical trial is greater than 18 years (>18 years), depending on local regulatory requirements. - Patients with recurrent or progressive metastatic gastric cancer involving the gastroesophageal junction or upper stomach who had received two or more lines of palliative chemotherapy before screening. For the dose determination portion, have at least one evaluable lesion and for the dose expansion portion, have at least one measurable lesion by RECIST v1.1. Eastern Cooperative Oncology Group (ECOG) performance status less than or equal to 1 (≦1). • If life expectancy is 12 weeks or more as of the time of registration. ● It can be administered orally. Patients with adequate organ function, including results of the last clinical laboratory evaluation performed within 1 week prior to study drug administration. For premenopausal women, a negative serum pregnancy test must be obtained within 4 weeks prior to study drug administration. Patients must also have a negative urine pregnancy test result prior to study drug administration on cycle day 1. Women who are infertile due to surgical sterilization or who are postmenopausal for more than 1 year are included without a serum and / or urine pregnancy test. ● Men and women of childbearing age must agree to use adequate contraception, as directed by the investigator, from the start of the first dose of study therapy until 6 months after the last dose. Effective contraception is defined as a method that has a low failure rate (i.e., <1% per year) when used consistently and correctly (e.g., implants, injections, combined oral contraceptives, or intrauterine devices). In the investigator's judgment, total abstinence also counts as acceptable contraception if it is consistent with the patient's lifestyle (cyclical abstinence [e.g., date cycle method, ovulation cycle method, symptomatic cycle method, postovulation method] and withdrawal are not acceptable contraception). • Willing and able to comply with all requirements of the research protocol, including the prohibitions and restrictions stated in the protocol.

[0132] Administration The compound of formula 1 is provided in 5 mg, 10 mg, 20 mg and 40 mg tablets. The compound of formula 1 and irinotecan are provided in a sterile, pale yellow, clear aqueous solution at a concentration of 20 mg / ml for dilution with 5% Dextrose (glucose) Injection, USP, or 0.9% Sodium Chloride Injection, USP, prior to intravenous (IV) infusion. The preferred diluent is 5% Dextrose Injection, USP.

[0133] The administration frequency and administration dose of the compound of Chemical Formula 1 and irinotecan are shown in Table 7.

[0134] The compound of Formula 1 was administered QD with at least 8 oz (approximately 240 mL) of water. Irinotecan was administered via IV infusion over 90 minutes after all study procedures were completed.

[0135] Efficacy evaluation Efficacy assessments included ORR assessed by RECIST v1.1, PFS, DCR, DOR, TTR, BOR, TTP, and OS.

[0136] Radiological tumor assessment included all sites of known or suspected disease by RECIST v1.1.

[0137] Survival follow-up was performed for all patients at least every 12 weeks after discontinuation of study therapy and continued until patient death or end of study, whichever occurred first.

[0138] Efficacy analysis ORR is defined as the proportion of patients achieving a confirmed partial or complete response as defined by RECIST v1.1. ORR analyses were performed in patients with measurable disease at baseline. Patients with inadequate post-baseline tumor evaluation at the time of ORR analysis were calculated as non-responders. ORR rates are provided for each treatment group, including the corresponding 95% Wilson confidence intervals (CIs).

[0139] PFS is defined as the time from initiation of study therapy to the date of radiologically confirmed disease progression (per RECIST v1.1) or death from any cause, whichever occurs first, as determined by the investigator.

[0140] DCR is defined as the proportion of patients who achieved BOR with confirmed complete response (CR), confirmed PR, or SD (minimum duration of SD is 6 weeks).

[0141] DOR is defined as the time from the first documented objective response (confirmed CR or confirmed PR) to the first documented objective disease progression or death from any cause, whichever occurs first. Censoring rules are similar to PFS.

[0142] TTR is defined as the time from the start of study therapy to the date of first documented objective response (confirmed CR or confirmed PR). Censoring rules are similar to those for PFS.

[0143] Confirmed BOR is defined as the best response assessed from the first dose of study drug until disease progression / recurrence or death. If a patient is assessed as a PR or CR responder at the final BOR assessment after meeting the initial response criteria at tumor assessment, the patient must be assessed as a PR or CR responder at the next sequential assessment. Patients with measurable disease and available tumor assessment results at baseline are included in the BOR analysis, and patients without tumor assessments after baseline are considered non-responders.

[0144] TTP is defined as the time from first administration of investigational drug to first disease progression. Patients without disease progression at study completion (including patients who died before disease progression) or patients lost to follow-up due to dropout were censored at the date of their last tumor assessment.

[0145] OS is defined as the time from first administration of study drug to death from any cause. Surviving patients will be censored at the date they were last known to be alive.

[0146] Experimental Example 5 Experimental Results The BOR assessment results for the patients are shown in Table 8. [Table 8]

[0147] Best overall response (BOR): The best overall response (smallest measured value) recorded from the start of treatment until disease progression / relapse occurs.

[0148] Complete response (CR): All observed tumors have completely disappeared and all pathological lymph nodes have been reduced to less than 10 mm.

[0149] Partial response (PR): If the sum of the diameters is reduced by at least 30%

[0150] Stable disease (SD): Not corresponding to CR, PR, or PD, i.e., the condition remains the same

[0151] Progressive disease (PD): An increase of at least 20% in the sum of diameters, an absolute increase of at least 5 mm in the sum of diameters, and the appearance of one or more new lesions

[0152] According to systematic studies, stage 3 gastric cancer is known to show unsatisfactory results in standard treatment, with an objective response rate (ORR = CR + PR) of 3% to 6% and a progression-free survival (PFS) of only 1.61 to 3.67 months. However, according to the experimental results, the combination of the pharmaceutical composition of the present invention showed an objective response rate of 33.33%, reaching 3 out of 9 patients, and a mean treatment period of more than 5 months (5.6 months as of the end of 2022), demonstrating remarkably excellent results.

Claims

1. A combination pharmaceutical composition for combined administration to prevent or treat cancer comprising: (i) a first pharmaceutical composition comprising, as an active ingredient, a compound represented by the following chemical formula 1 or a pharma- ceutically acceptable salt thereof; and 【Chemistry 1】 (ii) a second pharmaceutical composition comprising, as an active ingredient, at least one anti-cancer agent.

2. The combination according to claim 1, wherein the anti-cancer drug is selected from the group consisting of chemotherapy drugs, targeted therapy drugs, immunotherapy drugs and hormonal therapy drugs.

3. 3. The method of claim 2, wherein the chemotherapy drug is selected from the group consisting of alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors and mitotic inhibitors.

4. The combination according to claim 2, wherein the targeted therapeutic agent is selected from the group consisting of angiogenesis inhibitors, monoclonal antibodies, proteasome inhibitors and signal transduction inhibitors.

5. The combination of claim 2, wherein the immunotherapeutic agent is selected from the group consisting of checkpoint inhibitors, chimeric antigen receptor (CAR) T-cell therapeutic agents and cytokines.

6. The combination of claim 2, wherein the hormone therapy drug is selected from the group consisting of aromatase inhibitors, selective estrogen receptor modulators, estrogen receptor antagonists, luteinizing hormone releasing hormone agonists, antiandrogens and CYP17 inhibitors.

7. The combination according to claim 1 , wherein the anticancer drug is selected from the group consisting of irinotecan and oxaliplatin.

8. The combination according to claim 1 , wherein the anticancer drug is irinotecan.

9. The combination according to claim 1 , wherein the pharma- ceutically acceptable salt is a hydrochloride salt.

10. The combination according to claim 1 , wherein the combined pharmaceutical composition is administered in the form of a mixture of the first pharmaceutical composition and the second pharmaceutical composition.

11. The combination according to claim 1 , wherein the combined pharmaceutical composition is a first pharmaceutical composition and a second pharmaceutical composition, which are each formulated and administered simultaneously or sequentially.

12. The combination according to claim 1 , wherein the first pharmaceutical composition and the second pharmaceutical composition are each contained in the form of a separate dosage form.

13. The combination of claim 1 , wherein the first pharmaceutical composition is in an oral dosage form and the second pharmaceutical composition is in an injectable dosage form.

14. The combination of claim 1 , wherein the first pharmaceutical composition is administered once daily.

15. 2. The combination of claim 1, wherein the first pharmaceutical composition comprises about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, or about 50 mg of the compound represented by Formula I.

16. The method of claim 1 , wherein the first pharmaceutical composition is administered at about 1 mg to about 180 mg per dose.

17. 2. The combination of claim 1, wherein the first pharmaceutical composition is administered at about 10 mg, about 20 mg, about 40 mg, about 80 mg, about 120 mg or about 160 mg per dose.

18. The second pharmaceutical composition is about 30 mg / m ２ to about 120 mg / m ２ The combination according to claim 1, which is administered intravenously at

19. The second pharmaceutical composition is about 50 mg / m ２ , about 75mg / m ２ or about 100 mg / m ２ The combination according to claim 1, which is administered intravenously at

20. 2. The combination of claim 1, wherein the first pharmaceutical composition is administered in a cycle, wherein the cycle comprises administering the first pharmaceutical composition for about 14 days or about 21 days.

21. 2. The combination of claim 1, wherein the second pharmaceutical composition is administered in a cycle, wherein the cycle comprises administering the second pharmaceutical composition for about 14 days or about 21 days.

22. The combination of claim 1, wherein the second pharmaceutical composition is administered in a cycle, wherein the cycle is repeated at least once.

23. The combination according to claim 1, wherein the cancer is gastric cancer.

24. 10. A method for preventing or treating cancer, comprising co-administering a combination according to any one of claims 1 to 9 to a subject in need thereof.

25. 24. The method of claim 23, wherein the cancer is gastric cancer.

26. A kit for preventing or treating cancer, comprising the combination according to claim 1, and a first pharmaceutical composition, which is administered simultaneously, separately or sequentially.

27. 26. The kit of claim 25, wherein the first pharmaceutical composition is in a single dosage form and the second pharmaceutical composition is in a separate dosage form.

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