Pharmaceutical composition set for combination therapy for cancer prevention or treatment

Abstract

The present invention discloses a pharmaceutical composition set for combination therapy for the prevention or treatment of cancer. The pharmaceutical composition set of the present disclosure comprises: a first composition including a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt thereof, or a hydrate thereof; and a second composition including an immune anticancer agent. When administered in combination, the first and second compositions exhibit a significantly enhanced anticancer effect compared to when administered individually.

Description

Set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer

[0001] The present invention relates to a set of pharmaceutical compositions for combined administration for the prevention or treatment of cancer, and more particularly, to a set of pharmaceutical compositions in the form of a combination (set) composed of each individual composition, and to a set of pharmaceutical compositions that exhibit a synergistic effect through combined administration.

[0002] When there is a problem with the regulatory function of a cell, abnormal cells that should normally die do not die but instead proliferate excessively, invading surrounding tissues and organs to form a mass (lump) and destroy or deform the existing structure. A mass composed of undifferentiated cells that ignore order and proliferate without limit within the tissue can be defined as cancer. Recently, the incidence of cancer has been rapidly increasing compared to the past due to rapid industrial development, changes in the global ecosystem and eating habits, and various anticancer drugs are being researched and developed.

[0003] Among them, cisplatin, a compound composed of two chlorines and ammonia coordinated to a platinum atom, is one of the most widely used anticancer drugs in cancer treatment. Cancer cells are characterized by continuous proliferation due to an inability to suppress cell division. Cisplatin inhibits cell division by blocking the reproductive ability of cancer cells through changes in the DNA arrangement, thereby preventing cell proliferation and preventing further progression of the cancer. In other words, cisplatin exhibits anticancer properties. However, while cisplatin is an excellent anticancer drug effective against many cancers, it unfortunately exhibits various side effects, such as serious kidney damage, and is insoluble in water.

[0004] While cisplatin and other conventional anticancer agents effectively inhibit cancer cell growth, they can also be toxic to normal cells. Furthermore, the pharmacological effects of these agents vary depending on the type of cancer and the specific anticancer agent, and the side effects due to their toxicity can also vary, posing a significant challenge in cancer treatment. Therefore, there is a continuing need for research and development of new anticancer agents that can exhibit effective anticancer properties against specific cancers.

[0005] Meanwhile, the use of combination therapy, which is more effective than monotherapy in cancer treatment, is increasing. Combination therapy maximizes therapeutic efficacy by adding drugs with different mechanisms of action to established treatments with proven efficacy and safety. It is attracting attention for its potential to broaden treatment options and improve therapeutic efficacy for cancer patients. Consequently, numerous companies are developing treatments based on combination therapy.

[0006] One object of the present invention is to provide a set of different pharmaceutical compositions for the prevention or treatment of cancer, and to effectively suppress the growth of cancer cells while minimizing the side effects of existing anticancer drugs by administering them in combination.

[0007] In one aspect, the present invention provides a set of pharmaceutical compositions for combination administration for preventing or treating cancer, comprising: a first composition comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt of the compound, and one of a hydrate thereof; and a second composition comprising a first immuno-oncology agent.

[0008] [Chemical Formula 1]

[0009]

[0010] In one embodiment, the first immunotherapy agent may be any one selected from a PD-L1 inhibitor and a PD-1 inhibitor.

[0011] In one embodiment, the PD-L1 inhibitor may include at least one selected from the group consisting of atezolizumab, durvalumab, and avelumab.

[0012] In one embodiment, the PD-1 inhibitor may comprise Pembrolizumab.

[0013] In one embodiment, the first composition may be administered orally and the second composition may be administered intravenously.

[0014] In one embodiment, the second composition may further comprise a second immunotherapy agent having a different mechanism of action from the first immunotherapy agent.

[0015] In one embodiment, when the first immunotherapy agent is a PD-L1 inhibitor, the second immunotherapy agent may be a CTLA-4 inhibitor.

[0016] In one embodiment, the CTLA-4 inhibitor may comprise ipilimumab.

[0017] In one embodiment, a third composition comprising a targeted anticancer agent may be further included.

[0018] In one embodiment, the targeted anticancer agent is selected from the group consisting of Gefitinib, Erlotinib, Afatinib, Cetuximab, Trastuzumab, Pertuzumab, Trastuzumab emtansine, Trastuzumab deruxtecan, Crizotinib, Alectinib, Brigatinib, Bevacizumab, Sunitinib, Pazopanib, Sorafenib, Lenvatinib, Axitinib, Cabozantinib, Dasatinib, Nilotinib, Midostaurin, It may include one or more substances selected from the group consisting of Palbociclib, Ribociclib, Verzenio (Abemaciclib), and Olaparib.

[0019] In one embodiment, the third composition may be characterized as being administered orally or intravenously.

[0020] In one embodiment, the composition may further comprise a fourth composition comprising a cytotoxic anticancer agent.

[0021] In one embodiment, the cytotoxic anticancer agent is Cisplatin, Carboplatin, Oxaliplatin, Irinotecan, Mechlorethamine (nitrogen mustard), Cyclophosphamide, Ifosfamide, Melphalan, Chlorambucil, Thiotepa, Altretamine, Procarbazine, Busulfan, Carmustine (BCNU), Lomustine (CCNU), Dacarbazine (DTIC), Fluorouracil (5-FU), Capecitabine, Cytarabine, Gemcitabine, It may include at least one substance selected from the group consisting of Methotrexate, Mercaptopurine (6-MP), Vinblastine, Vincristine, Vinorelbine, Paclitaxel, Docetaxel, Etoposide, Topotecan, Dactinomycin, Doxorubicin, Daunorubicine, Mitomycine, Bleomycin, and L-asparaginase.

[0022] In one embodiment, the fourth composition may be characterized as being administered intravenously.

[0023] In one embodiment, the pharmaceutical composition set may be characterized by having anticancer activity against liver cancer cells or colon cancer cells by enhancing an immune response against liver cancer cells or colon cancer cells by engaging in an immune checkpoint pathway of immune cells in the body.

[0024] The set of pharmaceutical compositions for combination administration for cancer prevention or treatment according to the present invention is provided in the form of a set composed of pharmaceutical compositions having different mechanisms, thereby effectively suppressing the growth and proliferation of cancer cells while minimizing the side effects of existing anticancer drugs.

[0025] In particular, the pharmaceutical composition set of the present invention is designed to significantly increase anticancer efficacy through combined administration of the first and second compositions compared to their use alone, and to allow additional combination with targeted anticancer agents or cytotoxic anticancer agents as needed. This provides a novel anticancer treatment strategy that optimizes the interaction between the individual compositions to overcome drug resistance in cancer cells and enhance therapeutic efficacy while reducing toxicity to normal cells.

[0026] Figure 1 shows the chemical structure of the compound constituting the first composition of the present invention.

[0027] Figure 2 is a drawing showing the change in body weight of mice to evaluate toxicity by experimental group during the administration period in a liver cancer anticancer experiment using Tecentriq / ETN101.

[0028] Figure 3 is a drawing showing changes in tumor size to evaluate efficacy by experimental group during the administration period in a liver cancer anticancer experiment using Tecentriq / ETN101.

[0029] Figure 4 is a diagram showing the results of measuring serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels to evaluate liver function in each experimental group in a liver cancer anticancer experiment using Tecentriq / ETN101.

[0030] Figure 5 is a drawing showing the change in body weight of mice to evaluate toxicity by experimental group during the administration period in a liver cancer anticancer experiment using Keytruda / ETN101.

[0031] Figure 6 is a drawing showing changes in tumor size to evaluate efficacy by experimental group during the administration period in a liver cancer anticancer experiment using Keytruda / ETN101.

[0032] Figure 7 is a diagram showing the results of measuring serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels to evaluate liver function in each experimental group in a liver cancer anticancer experiment using Keytruda / ETN101.

[0033] Figure 8 is a drawing showing the change in body weight of mice to evaluate toxicity by experimental group during the administration period in a colon cancer anticancer experiment using PD-1 antibody / ETN101.

[0034] Figure 9 is a diagram showing the change in tumor volume for efficacy evaluation by experimental group during the administration period in a colon cancer anticancer experiment using PD-1 antibody / ETN101.

[0035] Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings. The present invention is susceptible to various modifications and variations, and thus specific embodiments are illustrated in the drawings and described in detail in the text. However, this is not intended to limit the present invention to a specific disclosed form, but rather to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention. Throughout the description of each drawing, similar reference numerals have been used to designate similar components.

[0036] The terminology used in this application is only used to describe specific embodiments and is not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this application, it should be understood that the terms "comprise" or "have" indicate the presence of a feature, step, operation, component, part, or combination thereof described in the specification, but do not preclude the possibility of the presence or addition of one or more other features, steps, operations, components, parts, or combinations thereof.

[0037] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0038]

[0039] The present invention relates to a set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer. Combination therapy is becoming increasingly important in cancer treatment to overcome the limitations of monotherapy and maximize therapeutic effects, and the present invention is based on this approach.

[0040] In this specification, the term "set of pharmaceutical compositions" refers to a set of pharmaceutical compositions in a dosage form composed of different pharmaceutical compositions, and refers to a collection of pharmaceutical compositions designed so that the individual pharmaceutical compositions are complementary to each other or exhibit synergistic effects through their respective mechanisms or actions. The present invention is characterized in that the set of pharmaceutical compositions is designed by considering the interaction between each composition so that the optimal therapeutic effect can be derived by co-administering the individual pharmaceutical compositions constituting the set of pharmaceutical compositions. This set of pharmaceutical compositions for co-administration has the advantage of reducing the side effects of a single anticancer agent, improving solubility in water, and maximizing the therapeutic effect by effectively targeting and killing specific cancer cells.

[0041] Hereinafter, the specific contents of the pharmaceutical composition set of the present invention will be described in detail through various examples.

[0042] (Example 1)

[0043] A set of pharmaceutical compositions according to one embodiment of the present invention comprises a first composition comprising a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt of the compound, and a hydrate thereof; and a second composition comprising a first immuno-oncology agent. Example 1 is designed to optimally function as a set of pharmaceutical compositions capable of maximizing the anticancer therapeutic effect by effectively suppressing the growth and proliferation of cancer cells while minimizing the side effects of existing anticancer agents through combined administration of the two compositions.

[0044] Figure 1 shows the chemical structure (chemical formula 1) of a compound constituting the first composition of the present invention. The compound represented by chemical formula 1 includes benzothiazole in which a benzene ring and a thiazole ring (a 5-membered heterocycle containing sulfur (S) and nitrogen (N)) are fused, and 2-phenylbenzothiazole in which a phenyl group is connected to the carbon (C) between sulfur (S) and nitrogen (N) of the thiazole ring serves as the core skeleton of the compound. In addition, it is a benzothiazole aniline structure in which a propionamide (-NHCOCH2CH3) group is connected to the para position of the phenyl group, and has a structure in which one hydrogen bonded to each of the α-carbon and β-carbon of the propionamide group is replaced with an amine group (amine, -NH2). By including the first composition having such a structure in a set of pharmaceutical compositions, solubility in water can be improved, biocompatibility can be enhanced, and the activity of the anticancer agent can be maintained.

[0045] The compound of the present invention includes an amine group. The first composition can be prepared in the form of a pharmaceutically acceptable salt (e.g., hydrochloride, sulfate, phosphate, acetate, etc.) through an acid-base reaction, and when such a salt form is formed, solubility can be increased and bioavailability can be improved. In addition, some compounds can exist in the form of a hydrate during the crystallization process, and such hydrate can be useful for increasing the stability of the compound and controlling drug release characteristics. Therefore, it should be understood that the first composition of the present invention includes not only the compound represented by Chemical Formula 1, but also a pharmaceutically acceptable salt or a hydrate thereof. The first composition can be formulated in the form of a powder, tablet, capsule, liquid, etc. In addition, it is suitable for oral administration and can be easily used in combination with other dosage forms.

[0046] The first composition of the present invention, when used in combination with existing immuno-oncology agents, targeted anticancer agents, and cytotoxic anticancer agents, can provide superior effects in the prevention and treatment of cancer compared to when used alone. The pharmaceutical composition set according to the present invention is characterized by co-administration of the first composition represented by Chemical Formula 1 and the second composition comprising the first immuno-oncology agent.

[0047] As used herein, "immunotherapy" refers to a substance that activates the body's immune system (e.g., T cells) to help effectively attack cancer cells. For example, the immunotherapy may be an immune checkpoint inhibitor, an immunocytotherapy, or an immunoviral therapy that induces changes in the immune system. Specifically, immune checkpoint inhibitors may include CTLA-4 inhibitors, PD-1 inhibitors, PD-L1 inhibitors, and the like, while immunocytotherapy may include T cells (e.g., CAR-T cells) and NK cells.

[0048] The immunotherapy agent may be formulated, for example, as a liquid, and administered intravenously. In one embodiment, when used in combination with the first composition, the first composition may be administered orally, and the immunotherapy agent may be administered intravenously. This administration strategy may help maintain therapeutic efficacy while enhancing patient convenience. However, the scope of the present invention is not limited to the above-described administration strategy, and those skilled in the art will be able to rationally select an appropriate administration regimen depending on the circumstances.

[0049] The combination administration strategy of the present invention maximizes the anticancer effects of immunotherapy and enhances therapeutic efficacy by inducing synergistic effects with the mechanisms of action of specific compounds. This combination blocks the immune evasion mechanisms of cancer cells, thereby increasing the response rate of immunotherapy and overcoming resistance that can develop with monotherapy. Furthermore, it can provide a safer and more effective cancer treatment while controlling side effects such as immune hypersensitivity.

[0050] The first immunotherapy agent may be any one selected from Tecentriq and Keytruda.

[0051] Tecentriq is an immunotherapy drug, the active ingredient of which is atezolizumab. It activates the immune system, inducing immune cells to selectively attack cancer cells. It is typically administered intravenously.

[0052] Keytruda, developed by MSD, is an immunotherapy drug with the active ingredient pembrolizumab. It acts as an immune checkpoint inhibitor, activating the patient's immune system to attack cancer cells. It is typically administered intravenously.

[0053] In addition, the second composition may further include, in addition to the first immunotherapy, a second immunotherapy having a different pharmacological mechanism from the first immunotherapy. For example, if the first immunotherapy has a mechanism of action that inhibits PD-L1 (e.g., atezolizumab, durvalumab, avelumab, etc.), the second immunotherapy may have a mechanism of action that inhibits CTLA-4 (e.g., ipilimumab, etc.). Since the types of the first and second immunotherapy agents are not limited to a specific mechanism in the present invention, and the first composition is used in combination, when a treatment regimen that combines immunotherapy agents with different mechanisms discovered to date is applied, it can be used together with the first composition of the present invention.

[0054] (Example 2)

[0055] A pharmaceutical composition set according to another embodiment of the present invention may include a first composition comprising a compound represented by Chemical Formula 1, a pharmaceutically acceptable salt of the compound, or a hydrate thereof; a second composition comprising a first immuno-oncology agent; and a third composition comprising a targeted anti-cancer agent. The descriptions of the first and second compositions are the same as those described above, and therefore, redundant descriptions are omitted.

[0056] As used herein, "targeted anticancer agent" refers to a substance that kills cancer cells by targeting specific genes, proteins, or signaling pathways that play a key role in cancer development. When used in combination with additional targeted anticancer agents, the drug effectively reaches cancerous tissue, selectively eliminating cancer cells while minimizing damage to normal cells.

[0057] The targeted anticancer drugs can be broadly classified into monoclonal antibodies, kinase inhibitors, and antibody-drug conjugates (ADCs). The monoclonal antibodies may include Erbituximab (Cetuximab), Herceptin (Trastuzumab), Perjeta (Pertuzumab), and Avastin (Bevacizumab), and the antibody-drug conjugates may include Kadcyla (Trastuzumab emtansine) and Enhertu (Trastuzumab deruxtecan). Additionally, kinase inhibitors may include Gefitinib, Erlotinib, Afatinib, Crizotinib, Alectinib, Brigatinib, Sunitinib, Pazopanib, Sorafenib, Lenvatinib, Axitinib, Cabozantinib, Dasatinib, Nilotinib, Midostaurin, Palbociclib, Ribociclib, Abeciclib, and Olaparib. Preferably, the targeted anticancer agent may be, but is not necessarily limited to, Bevacizumab.

[0058] Avastin is the brand name for bevacizumab, and can be used alone or in combination with other drugs to treat cervical, colon, lung, kidney, ovarian, uterine, peritoneal, and glioblastoma (a type of brain tumor). Bevacizumab is a monoclonal antibody that binds to vascular endothelial growth factor (VEGF) and inhibits the formation of blood vessels necessary for tumor growth, making it an antiangiogenic agent.

[0059] The third composition containing the targeted anticancer agent can be administered intravenously or orally. Therefore, when the first, second, and third compositions of the present invention are administered together, the first composition can be administered orally, the second composition can be administered intravenously, and the third composition can be administered intravenously or orally. Furthermore, those skilled in the art can adjust the administration method for each composition as needed.

[0060] (Example 3)

[0061] A pharmaceutical composition set according to another embodiment of the present invention may include a first composition comprising a compound represented by the following Chemical Formula 1, a pharmaceutically acceptable salt of the compound, and one of a hydrate thereof, a second composition comprising a first immuno-oncology agent, and a fourth composition comprising a cytotoxic-oncology agent. The descriptions of the first and second compositions are the same as those described above, and therefore, redundant descriptions are omitted.

[0062] In this specification, “cytotoxic anticancer agent” means a substance that kills cancer cells by interfering with the DNA and RNA synthesis process and mitosis of rapidly proliferating cancer cells or by having a detrimental effect on the DNA molecules themselves. For example, the cytotoxic anticancer drugs include cisplatin, carboplatin, oxaliplatin, irinotecan, mechlorethamine (nitrogen mustard), cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, carmustine (BCNU), lomustine (CCNU), dacarbazine (DTIC), fluorouracil (5-FU), capecitabine, cytarabine, gemcitabine, It may be at least one selected from the group consisting of Methotrexate, Mercaptopurine (6-MP), Vinblastine, Vincristine, Vinorelbine, Paclitaxel, Docetaxel, Etoposide, Topotecan, Dactinomycin, Doxorubicin, Daunorubicine, Mitomycine, Bleomycin, and L-asparaginase.

[0063] Preferably, a therapy comprising a cytotoxic anticancer agent selected from these may be used in combination with the first and second compositions. For example, it may be used with FOLFOX, FOLFIRI, and CapeOx regimens comprising irinotecan and 5-FU.

[0064] The fourth composition containing the cytotoxic anticancer agent can be administered intravenously. Therefore, when the first, second, and fourth compositions of the present invention are administered together, the first composition can be administered orally, while the second and fourth compositions can be administered intravenously. Furthermore, those skilled in the art can adjust the administration method for each composition as needed.

[0065] The composition set according to an embodiment of the present invention has a mechanism that activates an immune response by acting on the immune checkpoint pathway of immune cells in the body, and specifically targets liver cancer cells or colon cancer cells to promote immune attack. Generally, liver cancer cells or colon cancer cells utilize immune evasion mechanisms to avoid immune cell attacks. However, the composition set of the present invention modulates the immune checkpoint, thereby increasing the anticancer activity of immune cells and inducing the effective elimination of liver cancer cells or colon cancer cells.

[0066] In particular, patients with liver cancer or colon cancer tend to have high PD-L1 / PD-1 expression, making them a target for immune checkpoint inhibitors. Furthermore, existing single-agent immunotherapy often fails to achieve sufficient therapeutic effects. The composition set of the present invention was designed to take into account the immunological characteristics of liver cancer or colon cancer, and can provide a combination treatment strategy that optimizes the immune response in liver cancer or colon cancer, thereby achieving a more potent anticancer effect.

[0067] (Synthesis method of the first composition)

[0068] Based on the compound and complex synthesis method disclosed in registered patent invention 10-2304622, the first composition according to the present invention was prepared by applying the same or similar reaction conditions and procedures.

[0069] The compound synthesized in this way is named '2,3-diammoniumtrifluoroacetate-N-(4-benzothiazol-2-yl-phenyl)-propionamide', and is hereinafter abbreviated as 'ETN101'.

[0070] (Animal testing methods)

[0071] The experimental animals used in the humanized mouse liver cancer model combination therapy (Tecentriq / ETN101 and Keytruda / ETN101) experiment utilized in the evaluation example of the present invention are specific pathogen free (SPF) mice of the NOG (NOD / Shi-scid / IL-2Rγnull) strain. NOG mice are immunocompromised animals strained by backcrossing NOD / scid mice with IL-2Rγ gene knockout mice, and are deficient in T cells, B cells, and NK cells, making xenogeneic cell transplantation easy and a species widely used for producing humanized mice. To generate humanized mice in this experiment, human peripheral blood mononuclear cells (PBMCs) were purchased from Lonza (2C-2702, Lonza, 23TL198510) and were cultured at 2.5 × 10 in PBS (Gibco, 10010-023). 7 After adjusting the concentration to 10 cells / mL, 200 μl was intravenously injected per NOG mouse.

[0072] Meanwhile, in the mouse colon cancer model experiments of the present invention, BALB / c mice were used. Since the colon cancer cell line used in the experiments was derived from BALB / c mice, BALB / c mice of the same strain were used in the experiments to maintain immunological compatibility and increase experimental reliability.

[0073] (Cell culture and transplantation methods)

[0074] The cell culture conditions used in the evaluation examples of the present invention utilized culture media and environmental conditions optimized to the characteristics of the cell lines.

[0075] HepG2, a human hepatocellular carcinoma cell line, was cultured in a medium containing 10% FBS (Gibco, 16000-044) and 1% Penicillin / Streptomycin (Gibco, 15140-122) in MEM (Gibco, 11095-080). Cell transplantation was performed using PBS at a concentration of 2.5X10 HepG2 cells. 7 cells / mL, and 200 μl of PBS and Matrigel (Corning, 354234) were mixed in a 1:1 ratio and transplanted subcutaneously into humanized mice.

[0076] CT26, a mouse colon cancer cell line, was cultured in RPMI-1640 (Gibco) supplemented with 10% FBS (Corning) and 1% Penicillin / streptomycin (Gibco). Cell transplantation was performed using HBSS at a concentration of 5X10 CT26 cells. 6 cells / mL, and HBSS and Matrigel (Corning) were mixed in a 1:1 ratio and transplanted subcutaneously into the right flank of BALB / c mice that had undergone hair removal the previous day (100 μl each).

[0077] All cells were cultured in a CO2 incubator (Panasonic) at 37°C and 5% CO2 to maintain physiological conditions and provide an optimal growth environment. These culture conditions were established to meet the physiological needs of each cell line and ensure experimental reproducibility and reliability.

[0078] (Data analysis method)

[0079] Statistical analysis of data for liver cancer treatment was performed using SPSS 10.1. Data were expressed as mean ± standard deviation (SD), and one-way ANOVA was applied as the analysis method, followed by Tukey's post hoc test for multiple comparisons (*: p < 0.05, **: p < 0.01 and ***: p < 0.001 vs. G1, #: p < 0.05, ##: p < 0.01, ###: p < 0.001 vs. G2).

[0080] (Evaluation Example 1 - Tecentriq / ETN101 combination liver cancer anticancer trial)

[0081] Tumor volumes formed 7 days after HepG2 cell transplantation were measured. Group separation was performed by random assignment based on tumor volume. The efficacy evaluation experiment of the compound according to the present invention (hereinafter, ETN101) and Tecentriq was divided into experimental groups as shown in Table 1 below.

[0082] Group Sex Number of individuals (N) Cell line (number of cells / animal) Inoculation route Treatment Dose (mg / kg) Administration route G1 Male 7 HepG2 (5X10 6 )Subcutaneous injection (SC)Vehicle-oral administration (PO)G27Tecentriq(T)10Intravenous injection (IV)G37ETN10120Oral administration (PO)G47T / ETN10110 / 20Intravenous injection / oral administration (IV / PO)

[0083] In the efficacy evaluation trial of ETN101 and Tecentriq, ETN101 was dissolved in distilled water (Invitrogen, 10977-015) and administered orally daily for 14 days at the dosages specified in Table 1. Tecentriq was diluted with phosphate-buffered saline (PBS) and administered intravenously (IV) twice a week at the dosages specified in Table 1. The administration volume of each drug was set at 10 mL / kg. In the liver cancer treatment trial, the body weight of each experimental group was measured twice a week from the first day of administration (Day 0), and the weight change was observed until the end of the experiment. During the experimental period, a total of two mice in the Tecentriq alone administration group (G2) died. The body weight change (%) was calculated using the following formula.

[0084] Body weight (%) = (body weight / body weight on the day of starting administration) X 100

[0085] Figures 2a and 2b show the body weight changes by experimental group during the administration period. Compared with the excipient control group (G1), the average body weight of the Tecentriq monotherapy group (G2) significantly increased from day 3 to day 7 after the start of administration (p < 0.01 vs. G1), but returned to a level similar to that of the excipient control group (G1) over time. Meanwhile, compared with the excipient control group (G1), no statistically significant body weight changes were observed in the ETN101 monotherapy group (G3) and the Tecentriq / ETN101 combination therapy group (G4).

[0086] To evaluate the anticancer effect of each experimental group on liver cancer, tumor size was measured. Tumor size of the experimental group was measured twice a week from the start of administration (Day 0). Calipers were used to record the short axis (A) and long axis (B) of the tumor, and the tumor volume (mm) was calculated using the formula below. 3 ) was produced.

[0087] Tumor size (mm) 3 ) = [A(mm)] 2 XB(mm) X 0.5

[0088] Figures 3a and 3b show changes in tumor size by experimental group. On the 14th day after the start of administration, compared to the vehicle control group (G1), tumor size was reduced by 35.0% in the ETN101 monotherapy group (G3) (p < 0.05 vs. G1), and by 43.9% in the Tecentriq / ETN101 combination group (G4) (p < 0.01 vs. G1). In particular, the tumor growth inhibition effect was more pronounced when Tecentriq and ETN101 were administered together than when administered alone.

[0089] Figure 3c provides images that allow visual confirmation of tumor size for each individual experimental group. In the control group (G1), large, protruding tumors are clearly observed in the flank region, whereas in the ETN101 monotherapy group (G3) and the Tecentriq / ETN101 combination therapy group (G4), tumor size is noticeably reduced.

[0090] Anesthesia was induced by intraperitoneal injection of Zoletil™ (50 mg / kg, Virbac Laboratories, 06516) and Rompun (10 mg / kg, Elanco Animal Pharmaceuticals Korea, 192492). The animals were euthanized by laparotomy, blood collection from the inferior vena cava, and exsanguination. The liver, kidney, and tumor were excised, and the tumor weight was measured. In addition, blood was collected, serum was separated, and blood biochemical analysis (AST / ALT) was performed.

[0091] Figure 3d provides an image that can confirm the size of the tumors extracted by each experimental group after autopsy, and Figures 3e and 3f show the results of the tumor weights measured after autopsy. Compared to the vehicle control group (G1), the average tumor weight was reduced by approximately 47% in the Tecentriq monotherapy group (G2), whereas the average tumor weight was reduced by 54.3% in the ETN101 monotherapy group (G3) (p < 0.05 vs. G1), and the Tecentriq / ETN101 combination therapy group (G4) showed a higher anticancer effect with a 66.1% reduction (p < 0.01 vs. G1).

[0092] Figures 4a and 4b show the results of blood biochemical analysis of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and Figure 4c summarizes these results in numerical form. Generally, higher AST and ALT levels indicate more severe liver damage. In the excipient control group (G1) and the Tecentriq monotherapy group (G2), AST and ALT levels were abnormally elevated in certain individuals, suggesting liver damage in some individuals or the possibility of hepatotoxicity due to the experimental drug.

[0093] The results of this evaluation example 1 suggest that when the compound (ETN101) according to the present invention is administered in combination with Tecentriq, it can effectively suppress the growth of liver cancer while minimizing side effects due to cell damage, and this has the potential to exhibit a synergistic effect compared to single administration.

[0094] (Evaluation Example 2 - Keytruda / ETN101 combination liver cancer anticancer trial)

[0095] Seven days after HepG2 cell transplantation, the tumor volume was measured and randomly assigned to experimental groups based on this data. The efficacy evaluation trial for ETN101 and Keytruda was conducted by organizing the experimental groups according to Table 2.

[0096] Group Sex Number of individuals (N) Cell line (number of cells / animal) Inoculation route Treatment Dose (mg / kg) Administration route G1 Male 7 HepG2 (5X10 6 ) Subcutaneous injection (SC) Vehicle-oral administration (PO) G27 Keytruda (K) 10 Intravenous injection (IV) G37 K / ETN 10 110 / 40 Intravenous injection / oral administration (IV / PO) G47 K / ETN 10 110 / 82 Intravenous injection / oral administration (IV / PO) G57 ETN 10 140 Oral administration (PO) G67 ETN 10 182 Oral administration (PO)

[0097] In the efficacy evaluation trial of ETN101 and Keytruda, ETN101 was dissolved in distilled water and administered orally daily for 12 days at the dosages specified in Table 2. Keytruda was diluted with phosphate-buffered saline (PBS) and administered intravenously three times a week (at 2-3 day intervals) for a total of five doses. The total administered volume was set to 10 mL / kg, and the administered dose was adjusted based on the most recently measured body weight. The body weight of the experimental group for liver cancer treatment was measured twice a week from the start of administration (Day 0), and the body weight change was observed until the end of the experiment. The body weight change (%) was calculated using the following formula.

[0098] Body weight (%) = (measured body weight / body weight on the day of starting administration) × 100

[0099] During the experimental period, a total of two animals (G2-1, G3-3) died, and the body weight of the dead animals was confirmed to have decreased by more than 10% compared to the day of initiation of administration. Figures 5a and 5b show the change in body weight by experimental group. Compared to the vehicle control group (G1), the average body weight in the Keytruda monotherapy group (G2) significantly increased on the 4th day after initiation of administration (p < 0.05 vs. G1), but thereafter returned to a level similar to that of the vehicle control group (G1).

[0100] In addition, in the Keytruda / ETN101 low-dose administration group (G3), there was a decrease of approximately 6.7% on the 4th day after the start of administration (p < 0.01 vs. G1), and in the ETN101 low-dose only administration group (G5), there was a decrease of approximately 7.5% on the 7th day (p < 0.05 vs. G1), but body weight was recovered over time and showed a level similar to that of the vehicle control group (G1).

[0101] Meanwhile, in the Keytruda / ETN101 high-dose group (G4), the average body weight decreased by approximately 8% from day 4 to day 7 after the start of administration (p < 0.05 vs. G1), but no statistically significant difference was observed compared to the vehicle control group (G1) on day 11 after the start of administration.

[0102] To evaluate the anticancer effect of each experimental group on liver cancer, tumor size was measured. The tumor size of the experimental group was measured twice a week from the start of administration (Day 0). The short axis (A) and long axis (B) of the tumor were recorded using calipers, and the tumor volume (mm) was calculated using the following formula. 3 ) was produced.

[0103] Tumor size (mm) 3 ) = [A(mm)] 2 XB(mm) X 0.5

[0104] Figures 6a and 6b show the changes in tumor size by experimental group. Compared with the excipient control group (G1), the tumor size in the Keytruda monotherapy group (G2) significantly decreased from day 7 to day 11 after the start of administration (p < 0.05 vs. G1 at day 7 & 11). In addition, the tumor size in the Keytruda / ETN101 low-dose combination group (G3) and the Keytruda / ETN101 high-dose combination group (G4) significantly decreased from day 4 to day 11 after the start of administration (p < 0.01 vs. G1 at day 4, p < 0.001 vs. G1 at days 7 & 11).

[0105] In the low-dose ETN101 single-administration group (G5), the tumor size significantly decreased from day 7 to day 11 after the start of administration (p < 0.001 vs. G1 at day 7 & 11), and in the high-dose ETN101 single-administration group (G6), a significant decrease was confirmed from day 4 to day 11 after the start of administration (p < 0.01 vs. G1 at day 4, p < 0.001 vs. G1 at day 7 & 11).

[0106] On the 11th day after the start of administration, compared to the excipient control group (G1), the tumor size was reduced by 25% in the Keytruda monotherapy group (G2), and by 72.5% and 75.0% in the Keytruda / ETN101 low-dose combination group (G3) and high-dose combination group (G4), respectively. In addition, a 58.7% decrease in tumor size was confirmed in the ETN101 low-dose monotherapy group (G5), and a 73.1% decrease in the ETN101 high-dose monotherapy group (G6).

[0107] Meanwhile, compared to the Keytruda monotherapy group (G2), the Keytruda / ETN101 low-dose combination therapy group (G3) showed a 63.3% tumor reduction on day 11 after the start of administration (p < 0.001 vs. G2), and the Keytruda / ETN101 high-dose combination therapy group (G4) showed a significant decrease in tumor size from day 7 to day 11 after the start of administration (p < 0.01 vs. G2 at day 7, p < 0.001 vs. G2 at day 11). In particular, the Keytruda / ETN101 high-dose combination therapy group (G4) showed a 66.7% tumor reduction rate on day 11 after the start of administration compared to the Keytruda monotherapy group (G2).

[0108] Anesthesia was induced by intraperitoneal injection of Zoletil™ (50 mg / kg, Virbac Laboratories, 06516) and Rompun (10 mg / kg, Elanco Animal Pharmaceuticals Korea, 192492). The animals were euthanized by laparotomy, blood collection from the inferior vena cava, and exsanguination. The liver, kidney, and tumor were excised, and the tumor weight was measured. In addition, blood was collected, serum was separated, and blood biochemical analysis (AST / ALT) was performed.

[0109] Figures 6c, 6d, and 6e show the size and weight of the excised tumors in each experimental group. Compared to the excipient control group (G1), the average tumor weight in the Keytruda monotherapy group (G2) was reduced by approximately 17.4%, but no statistically significant difference was confirmed. On the other hand, the tumor weight was reduced by 87.3% in the Keytruda / ETN101 low-dose combination group (G3) (p < 0.001 vs. G1), and by 93.8% in the Keytruda / ETN101 high-dose combination group (G4) (p < 0.001 vs. G1). Additionally, in the low-dose ETN101 monotherapy group (G5), the average tumor weight was reduced by 87.8% (p < 0.05 vs. G1), and in the high-dose ETN101 monotherapy group (G6), the average tumor weight was reduced by 92.8% (p < 0.001 vs. G1).

[0110] Compared to the Keytruda monotherapy group (G2), the mean tumor weight was reduced by 84.7% in the Keytruda / ETN101 low-dose combination group (G3) (p < 0.001 vs. G2), and by 92.5% in the Keytruda / ETN101 high-dose combination group (G4) (p < 0.001 vs. G2), showing that combination therapy was more effective in reducing tumors.

[0111] Meanwhile, Figures 7a and 7b show the results of blood biochemical analysis of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and Figure 7c summarizes these results in numerical form. Generally, higher AST and ALT levels indicate more severe liver damage. In the excipient control group (G1) and the Keytruda monotherapy group (G2), AST and ALT levels were abnormally elevated in certain individuals, suggesting liver damage in some individuals or the possibility of hepatotoxicity due to the experimental drug.

[0112] The results of this evaluation example 2 suggest that when the compound (ETN101) according to the present invention is administered in combination with Keytruda, it can effectively suppress the growth of liver cancer while minimizing side effects due to cell damage, which suggests the possibility of a synergistic effect compared to monotherapy.

[0113] (Evaluation Example 3 - PD-1 Antibody / ETN101 Combination Liver Cancer Anticancer Experiment)

[0114] After CT26 cell transplantation, the tumor volume averaged 75-80 mm 3 When the tumor volume was reached, the experimental groups were separated using a random distribution method. At that point, the groups were randomly distributed based on the tumor volume, and the experimental groups were composed according to Table 3. During the experimental period, individual identification was performed using the ear-punch method, and identification cards for each group were attached to the breeding box for management.

[0115] Group Sex Number of individuals (N) Cell line (number of cells / animal) Inoculation route Treatment Dose (mg / kg) Administration route G1 Male 10 CT26 (5X10 5 ) Subcutaneous injection (SC) Vehicle-Oral administration (PO) G210 ETN10 182 Oral administration (PO) G310 Anti-mouse PD-110 Intraperitoneal injection (IP) G410 ETN101 + Anti-mouse PD-182 / 10 Oral administration / Intraperitoneal injection (PO / IP)

[0116] The compound (ETN101) according to the present invention was dissolved in sterile distilled water and administered orally daily for 18 days at the dosages specified in Table 3. The anti-mouse PD-1 antibody used as a mouse immunotherapy agent was diluted with PBS and administered intraperitoneally twice a week at the dosages specified in Table 3, for a total of 5 administrations. The total administered volume was set to 10 mL / kg. Statistical comparisons of experimental data for colorectal cancer treatment were analyzed using the GraphPad Prism 9 program. First, a normality test between groups was performed, and then the significance between the experimental groups was examined using the ANOVA test or the Kruskal-Wallis test, and the significance test criterion was set at p < 0.05.

[0117] Body weights of the experimental groups for colon cancer treatment were measured once on the first day of administration and then twice a week thereafter, and were also recorded on the day of autopsy. Figure 8 graphically depicts the weight changes in each group during the experimental period. Throughout the experiment, mice in all experimental groups maintained normal body weights, and no moribundity or behavioral abnormalities were observed due to drug administration.

[0118] To evaluate the anticancer effect of each experimental group on colon cancer, tumor size was measured. Tumor size was measured once the day before group separation and then twice a week thereafter to calculate tumor volume. Tumor size (mm) 3 ) was measured using calipers for the short axis (A) and long axis (B) of the tumor, and then calculated using the formula below.

[0119] Tumor size (mm) 3 ) = [A(mm)] 2 XB(mm) X 0.5

[0120] Figure 9a is a graph showing the change in tumor volume by experimental group. From day 7 after administration of the experimental substance, compared to the vehicle control group (G1), the ETN101 administration group (G2), the anti-mouse PD-1 administration group (G3), and the ETN101 + anti-mouse PD-1 combination administration group (G4) all showed a tendency for the tumor volume increase rate to decrease. In particular, a statistically significant difference was confirmed from day 14, and the ETN101 + anti-mouse PD-1 combination administration group (G4) showed the best anticancer effect.

[0121] Comparison of tumor volumes at the end of the experiment showed that the ETN101 administration group (G2) and the ETN101+Anti-mouse PD-1 combination administration group (G4) showed a statistically significant decrease in tumor volume increase rate compared to the vehicle control group (G1).

[0122] After the experiment, autopsies were performed, and organ toxicity was visually assessed. Tumors were removed and weighed. Half were used for efficacy evaluation using flow cytometry, and the other half for efficacy analysis using Western blot. Additionally, spleens were removed and used for efficacy evaluation using flow cytometry.

[0123] Figures 9b and 9c show tumor size and weight after autopsy for each experimental group. Tumor weight measurements showed a tendency for tumor weight to decrease in all experimental groups compared to the vehicle control group (G1), and statistically significant decreases were observed, particularly in the anti-mouse PD-1 administration group (G3) and the ETN101+anti-mouse PD-1 combination administration group (G4).

[0124] The results of this evaluation example 3 suggest that when the compound (ETN101) according to the present invention is administered in combination with a PD-1 antibody, it can effectively suppress the growth of colon cancer and enhance the immune response to maximize the anticancer effect.

[0125]

[0126] The pharmaceutical composition set of the present invention possesses excellent anticancer activity and, based on this, can be utilized as an effective anticancer agent. In particular, the compounds and complexes of the present invention can selectively target specific cancer cells based on their structural characteristics, thereby exhibiting high anticancer activity. Furthermore, this selective targeting effect can suppress the growth of abnormal cancer cells while minimizing the impact on normal cells, thereby reducing side effects compared to existing anticancer agents. Therefore, the pharmaceutical composition set of the present invention is expected to be widely utilized as an anticancer agent based on precision medicine targeting specific cancer cells.

[0127]

[0128] Although the present invention has been described above with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various modifications and changes may be made to the present invention without departing from the spirit and scope of the present invention as set forth in the claims below.

Claims

1. A first composition comprising a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt of the compound, and any one of the hydrates thereof; and Comprising a second composition comprising a first immunotherapy agent, Set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer: [Chemical Formula 1] 2. In paragraph 1, The first immunotherapy agent is one selected from among a PD-L1 inhibitor and a PD-1 inhibitor. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

3. In paragraph 2, The PD-L1 inhibitor comprises at least one selected from the group consisting of atezolizumab, durvalumab, and avelumab. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

4. In paragraph 2, The above PD-1 inhibitor includes pembrolizumab. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

5. In paragraph 1, The first composition is for oral administration, and the second composition is for intravenous administration, characterized in that A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

6. In paragraph 5, The second composition further comprises a second immunotherapy agent having a different mechanism of action from the first immunotherapy agent. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

7. In paragraph 6, If the first immunotherapy agent is a PD-L1 inhibitor, the second immunotherapy agent is a CTLA-4 inhibitor. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

8. In paragraph 7, The CTLA-4 inhibitor includes ipilimumab. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

9. In paragraph 1, Further comprising a third composition comprising a targeted anticancer agent, A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

10. In paragraph 9, The targeted anticancer drugs include Iressa (Gefitinib), Tarceva (Erlotinib), Giotrif (Afatinib), Erbitux (Cetuximab), Herceptin (Trastuzumab), Perjeta (Pertuzumab), Kadcyla (Trastuzumab emtansine), Enhertu (Trastuzumab deruxtecan), Xalico (Crizotinib), Alecenza (Alectinib), Alunbrig (Brigatinib), Avastin (Bevacizumab), Suten (Sunitinib), Votrient (Pazopanib), Nexavar (Sorafenib), Lenvima (Lenvatinib), Inlyta (Axitinib), Cabometyx (Cabozantinib), Sprycel (Dasatinib), Tasigna (Nilotinib), Rydap (Midostaurin), Ibrance (Palbociclib), Containing at least one substance selected from the group consisting of Kisqali (Ribociclib), Verzenio (Abemaciclib) and Lynparza (Olaparib); A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

11. In paragraph 9, The third composition is characterized in that it is for oral administration or intravenous administration. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

12. In paragraph 1, Further comprising a fourth composition comprising a cytotoxic anticancer agent, A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

13. In paragraph 12, The above cytotoxic anticancer drugs include cisplatin, carboplatin, oxaliplatin, irinotecan, mechlorethamine (nitrogen mustard), cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, carmustine (BCNU), lomustine (CCNU), dacarbazine (DTIC), fluorouracil (5-FU), capecitabine, cytarabine, gemcitabine, Containing at least one substance selected from the group consisting of Methotrexate, Mercaptopurine (6-MP), Vinblastine, Vincristine, Vinorelbine, Paclitaxel, Docetaxel, Etoposide, Topotecan, Dactinomycin, Doxorubicin, Daunorubicine, Mitomycine, Bleomycin, and L-asparaginase. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

14. In paragraph 12, The fourth composition is characterized in that it is for intravenous administration. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

15. In paragraph 1, The above pharmaceutical composition set is characterized in that it has anticancer activity against liver cancer cells or colon cancer cells by strengthening the immune response against liver cancer cells or colon cancer cells by involving the immune checkpoint pathway of immune cells in the body. A set of pharmaceutical compositions for combination administration for the prevention or treatment of cancer.

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