Pharmaceutical composition

Abstract

Providing cancer treatment or preventive agents. [Solution] A therapeutic or prophylactic agent for cancer comprising, for example, a compound represented by the following formula or a pharmaceutically acceptable salt thereof. TIFF2026116034000016.tif64170

Description

[Technical Field]

[0001] This invention relates to a novel therapeutic or prophylactic agent for cancer that targets the riboflavin metabolic pathway. [Background technology]

[0002] Among cancers, there are refractory cancers for which there are few effective treatment options. Pancreatic cancer, in particular, is a representative example of a refractory cancer, and the number of patients and deaths is certain to increase in the future, making the development of new therapeutic drugs for refractory cancers an urgent welfare issue. The inventors recently created and used flies (4-hit flies) that mimic the genotype (all mutations in KRAS, TP53, CDKN2A, and SMAD4; hereinafter referred to as 4-hit) of pancreatic cancer patients with the worst prognosis. They identified the riboflavin metabolic pathway and MEK kinase as novel therapeutic targets for pancreatic cancer, and identified a combination of the riboflavin metabolic pathway inhibitor roseoflavin (RoF) and the MEK inhibitor trametinib (Tr) (hereinafter referred to as RT) as a novel candidate for pancreatic cancer (US Patent 11,925,646). However, the inventors also found that this combination causes serious side effects, including death in the administered mice. [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] U.S. Patent No. 11,925,646 [Overview of the project] [Problems that the invention aims to solve]

[0004] In light of the above circumstances, there was a need to develop an anticancer drug that would not cause side effects and would be effective against cancer. [Means for solving the problem]

[0005] Therefore, the present inventor has conducted intensive studies to solve the above problems and has completed the present invention. That is, the present invention is as follows. [1] The following formula I: [Chemical formula] [In the formula, R1 represents [Chemical formula] and R2 represents -OH, -OR , , or -O-CO-R 21 (where R 21 represents an alkyl group), a group represented by the following formula 11: [Chemical formula] (In the formula, R 31 and R 32 each independently represents a hydrogen atom, an alkyl group, or -CO-R 21 ), or -O-CON(R 41 )(R 42 )(where R 41 and R 42 each independently represents a hydrogen atom or an alkyl group).) represents a group. ] A therapeutic or prophylactic agent for cancer, comprising a compound represented by . [2] The agent according to [1], wherein R2 is a hydroxy group or an amino group. [3] The compound represented by formula I is the following formula: [Chemical formula] The agent according to [1], which is represented by . [4] The agent according to [1], wherein the cancer is a digestive tract cancer, a respiratory cancer, a urinary cancer, a head and neck cancer, a breast cancer, or a blood cancer. [5] The agent described in [1], wherein the gastrointestinal cancer is pancreatic cancer, colorectal cancer, salivary gland cancer, or liver cancer. [6] A drug described in [1] for gastrointestinal cancer being pancreatic cancer. [7] A combination cancer therapy agent comprising one of the agents described in any one of items [1] to [3] and a MEK inhibitor. [8] The agent described in [7], wherein the MEK inhibitor is at least one selected from the group consisting of trametinib, binimetinib, selumetinib, mildametinib, cobimetinib, PD-334581, TAK-733, AZD8330, PD-198306, PD-98059, PD-0325901, U0126, Myrivetin, BI-847325, rifametinib, BI847325, GDC-0623, RGB-286638, Ro5126766, Isorhamnetin, CI-1040, APS-2-79, BIX02189, Hi-TOPK-032, pimasertib, PD318088, and Honokiol. [9] The agent described in [7] is a MEK inhibitor trametinib. [Effects of the Invention]

[0006] The present invention provides a preventive or therapeutic agent for cancer. The preventive or therapeutic agent of the present invention successfully created RoF derivatives, based on an idea that generates seeds with higher clinical utility. [Brief explanation of the drawing]

[0007] [Figure 1] This figure shows the results of suppressing the proliferation of pancreatic cancer cells with the combined use of roseoflavin and trametinib. [Figure 2] This figure shows the results of tumor growth suppression in a pancreatic cancer model mouse when roseoflavin and trametinib are used in combination. [Figure 3] This figure shows the Kd values ​​of the riboflavin derivatives RoF22 and RoF24 relative to RFK. [Figure 4]This figure shows the results of tumor growth suppression in a pancreatic cancer model mouse. [Figure 5] This figure shows the changes in body weight in a pancreatic cancer model mouse. [Figure 6] This figure shows the results of tumor growth suppression in a pancreatic cancer model mouse. [Figure 7] This figure shows the changes in body weight in a pancreatic cancer model mouse. [Modes for carrying out the invention]

[0008] The present invention relates to the following formula I: [ka] [In the formula, R1 is [ka] R2 represents -OH, -OR 21 , or -O-CO-R 21 (Here, R 21 represents an alkyl group. ) The group shown in formula 11: [ka] (In the formula, R 31 and R 32 Each of these independently comprises a hydrogen atom, an alkyl group, or -CO-R 21 This represents the group indicated by ), or -O-CON(R 41 )(R 42 )(Here, R 41 and R 42 Each of these independently represents either a hydrogen atom or an alkyl group. ) It represents a group indicated by ). This relates to a cancer treatment or preventive agent containing the compound indicated by [the specified compound].

[0009] 1. Overview The inventors first aimed to obtain derivatives with higher antitumor efficacy than roseoflavin (RoF), and created a group of RoF derivatives, then conducted compound screening by administering them to pancreatic cancer genotype model flies (4-hit flies). In this study, 4-hit flies were administered each RoF derivative in combination with trametinib (Tr), and it was found that the combination of RoF22 or RoF24 with Tr was more effective in increasing survival rates than RT. Furthermore, intermolecular interaction analysis was performed to investigate the interaction between these derivatives and riboflavin kinase (RFK), and it was revealed that both RoF22 and RoF24 bind to RFK more strongly than riboflavin.

[0010] Next, the inventors conducted a chemogenetic screening targeting all fly kinases to identify the more detailed mechanisms of action of RoF, RoF22, and RoF24. Specifically, RT was administered to 4-hit flies into which each of the fly kinase gene mutations had been introduced, and their survival rates were evaluated. As a result, it was found that inhibition of 10 kinases, including SRPK, significantly increased the survival rate of 4-hit flies that ingested RT. Furthermore, docking analyses were performed on the protein structures of these 10 kinases and their respective interactions with RoF, RoF22, and RoF24, yielding results suggesting that RoF22 and RoF24 bind to SRPK more strongly than RoF. From these results, the inventors concluded that RoF22 and RoF24 exert a higher effect than RoF by targeting SRPK in addition to RFK.

[0011] Furthermore, to investigate the usefulness of these derivatives in mammals, efficacy and toxicity tests were conducted using a mammalian pancreatic cancer model. First, the inventors created syngeneic pancreatic cancer model mice by subcutaneously transplanting KPC cell lines containing Kras activation and Trp53 inactivation into C57BL / 6J mice. Then, RoF22 and RoF24 were administered to these mice alone or in combination with Tr, and their tumor growth inhibitory effects were compared with those of RoF and RT. As a result, it was found that monotherapy with RoF22 and RoF24 inhibited tumor growth, and that the combination of each with Tr was more effective in inhibiting tumor growth than RT. Furthermore, the inventors conducted toxicity evaluations using mouse body weight as an indicator and found that the combination of RoF24 and Tr did not cause weight loss. These results indicate that RoF22 monotherapy, RoF24 monotherapy, and the combination of RoF24 and Tr are potential novel pancreatic cancer therapeutic agents with high clinical utility.

[0012] 2. Riboflavin derivatives The compound used in the present invention is a riboflavin derivative, represented by the following formula I. [ka]

[0013] In the formula, R1 is [ka] R2 represents -OH, -OR 21 , or -O-CO-R 21 (Here, R 21 represents an alkyl group. ) The group shown in formula 11: [ka] (In the formula, R 31 and R 32 Each of these independently comprises a hydrogen atom, an alkyl group, or -CO-R 21 This represents the group indicated by ), or -O-CON(R 41 )(R 42 )(Here, R 41 and R42 Each of these independently represents either a hydrogen atom or an alkyl group. ) represents the group indicated by ).

[0014] The group shown in formula 11 is an N-alkylamino group (-NH-R 21 )(R 21 ) represents an alkyl group. ), N-acylamino group (-NH-CO-R 21 )(R 21 ) represents an alkyl group. The alkyl group represents a linear or branched hydrocarbon group having 1 to 10 carbon atoms, and examples include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, octyl, nonyl, and decyl groups. In one embodiment of the present invention, R2 is a hydroxyl group or an amino group.

[0015] In a preferred embodiment of the present invention, the compound represented by formula I is, for example, the following: [ka] This is shown. The compound represented by formula I can be produced by known methods based on the description in Japanese Patent Publication No. 2011-530595, or by reacting 8-Chloro-8-demathylriboflavin (ACS Chem. Biol. 2018, 13, 2908-2919. Structure-Activity Relationship of Flavin Analogues That Target the Flavin Mononucleotide Riboswitch) with the target amine.

[0016] 3. Cancer The cancers targeted for treatment or prevention in this invention are not particularly limited, but examples include gastrointestinal cancers, respiratory cancers, urinary tract cancers, head and neck cancers, breast cancers, or hematological cancers. The specific cancer types (including classifications) within these cancers are as follows:

[0017] Gastrointestinal cancers: pancreatic cancer, colorectal cancer, salivary gland cancer, or liver cancer Urological cancers: Prostate cancer, bladder cancer, renal cell carcinoma, renal pelvis cancer, ureteral cancer, testicular cancer, penile cancer, etc. Head and neck cancers: nasal or paranasal sinus cancer, oral cancer, pharyngeal or laryngeal cancer, salivary gland cancer, thyroid cancer, etc.

[0018] Breast cancer: Non-invasive cancer, invasive cancer, Paget's disease, etc. Blood cancers: Leukemia (acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), malignant lymphoma (Hodgkin lymphoma, non-Hodgkin lymphoma), multiple myeloma (monoclonal gammaglobulinemia, asymptomatic myeloma, symptomatic myeloma)

[0019] In this invention, it is preferable to target gastrointestinal cancers, particularly pancreatic cancer.

[0020] 4. Cancer prevention or treatment agents and combination therapy agents, pharmaceutical compositions In the present invention, the compound represented by formula I may be used alone as a cancer preventive or therapeutic agent, or a pharmaceutically acceptable salt or hydrate thereof may be used. Furthermore, it can be formulated as a pharmaceutical composition by incorporating a known pharmaceutically acceptable carrier or the like.

[0021] Here, “treatment” means achieving some desired therapeutic effect, such as inhibiting the progression of a disease, whether in humans or animals (for example, in veterinary use, including pets such as dogs and cats, and livestock such as cattle, horses, and pigs), and includes, for example, slowing or stopping the rate of cancer progression, improving or curing the disease, or reducing or alleviating at least one symptom associated with or caused by cancer. Furthermore, "prevention" refers to achieving some desired preventive effect, such as preventing the development of cancer or providing protection against cancerous diseases, whether in humans or animals (for example, for the veterinary uses mentioned above). This includes, for example, preventing the worsening of all cancer symptoms or reducing the likelihood of cancer developing.

[0022] In the present invention, the pharmaceutically acceptable salts of the compound represented by formula I are not particularly limited and include, for example, salts with inorganic bases such as sodium salts, magnesium salts, potassium salts, calcium salts, and aluminum salts; salts with organic bases (e.g., methylamine, ethylamine, ethanolamine, etc.); salts with basic amino acids (lysine, ornithine, arginine, etc.); and ammonium salts. These salts may also be acid addition salts, and examples include acid addition salts with inorganic acids (hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, etc.), organic acids (formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, malic acid, tartaric acid, fumaric acid, succinic acid, lactic acid, maleic acid, citric acid, methanesulfonic acid, ethanesulfonic acid, etc.), and acidic amino acids (aspartic acid, glutamic acid, etc.).

[0023] Examples of pharmaceutically acceptable carriers include excipients, binders, disintegrants, lubricants, colorants, flavoring and odor-correcting agents, stabilizers, emulsifiers, absorption enhancers, surfactants, pH adjusters, preservatives, and antioxidants.

[0024] Furthermore, the administration form of the therapeutic agent, prophylactic agent, and pharmaceutical composition of the present invention is not particularly limited and can be administered orally or parenterally.

[0025] Examples of parenteral administration include injections (intravenous injection, intravenous drip infusion, subcutaneous injection, intradermal injection, intraperitoneal injection, or intramuscular injection). Injectable preparations can be prepared by adding diluents (e.g., glycols such as propylene glycol and polyethylene glycol, alcohols such as ethanol), suspensions, solubilizers, stabilizers, isotonic agents, preservatives, pH adjusters, buffers, etc., as needed. Injectable preparations can also be manufactured in a form that can be prepared immediately before use. That is, they can be prepared as a sterile solid composition by methods such as freeze-drying and dissolved in sterile distilled water for injection or other solvents before use. When administered by transdermal absorption as a patch, it is preferable to select a so-called free form that does not form salts. Injectable preparations can be prepared by conventional methods as intravenous drip infusion preparations, or as intravenous, subcutaneous, or intramuscular injection preparations.

[0026] Dosage forms used for oral administration include tablets, powders, granules, capsules, and syrups. Dosage forms used for parenteral administration include suppositories, injections, ointments, and poultices. When preparing oral formulations for oral administration, the active ingredient is combined with excipients, and optionally with binders, disintegrants, lubricants, colorants, flavoring agents, and odor-masking agents. The formulation can then be prepared by conventional methods as tablets, coated tablets, powders, granules, capsules, syrups, etc.

[0027] In the present invention, the effective dose of the compound represented by formula I, its pharmaceutically acceptable salt, or its hydrate varies depending on the severity of the symptoms, the patient's age, sex, weight, method of administration, timing of administration, interval between administrations, duration of administration, and properties of the formulation, but can be appropriately determined by a person skilled in the art.

[0028] For example, when administered orally, adults can be given doses in the following ranges per day: 0.01 mg / kg to 1000 mg / kg, 0.03 mg / kg to 1000 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 5 mg / kg to 250 mg / kg, 250 to 1000 mg / kg, etc. This can be administered once a day or in multiple doses (for example, three times a day).

[0029] For parenteral administration, the drug can be dissolved or suspended in a pharmaceutically acceptable carrier such as physiological saline or commercially available distilled water for injection to an appropriate concentration and administered as needed. For example, in the case of injectable preparations, adults can be administered in the following ranges per day: 0.01 mg / kg to 1000 mg / kg, 0.03 mg / kg to 1000 mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 5 mg / kg to 250 mg / kg, 250 to 1000 mg / kg, etc. This can be administered once a day or in multiple divided doses (for example, three times a day).

[0030] 5. Combination therapy agents for cancer Furthermore, in one embodiment of the present invention, a combination cancer therapy agent is provided, comprising a cancer preventive or therapeutic agent containing a compound represented by formula I and a MEK inhibitor. MEK inhibitors are drugs that suppress the proliferation of cancer cells by inhibiting the activation of MEK (MEK protein), which causes abnormal cell proliferation. Cancer cells damage normal cells and destroy tissues by repeatedly proliferating uncontrollably and metastasizing. When a mutation occurs in BRAF, a key factor (pathway) in cell proliferation signaling, the MEK and ERK pathways are abnormally activated, promoting the proliferation of cancer cells. Therefore, by using MEK inhibitors in combination, the proliferation of cancer cells can be suppressed.

[0031] Examples of MEK inhibitors, though not limited to them, include trametinib, binimetinib, selumetinib, mildametinib, cobimetinib, PD-334581, TAK-733, AZD8330, PD-198306, PD-98059, PD-0325901, U0126, Myrivetin, BI-847325, rifametinib, BI847325, GDC-0623, RGB-286638, Ro5126766, Isorhamnetin, CI-1040, APS-2-79, BIX02189, Hi-TOPK-032, pimasertib, PD318088, and Honokiol, which can be used alone or in combination of two or more. These MEK inhibitors can be obtained from companies such as MedChemExpress.

[0032] In this specification, “combination therapy” means therapy involving the use of a combination of the compound shown in Formula I and a MEK inhibitor. Therefore, “combination therapy” refers to a configuration in which the compound shown in Formula I and the MEK inhibitor are administered simultaneously or separately as part of the same overall treatment regimen. The respective dosages may differ, and they may be administered at the same time or at different times. Therefore, the combination of the compound shown in Formula I and the MEK inhibitor may be administered together as the same pharmaceutical formulation, or separately as different pharmaceutical formulations, consecutively or simultaneously. When administered simultaneously in the same formulation, it constitutes a single formulation, and when administered simultaneously in different pharmaceutical formulations, it may constitute multiple formulations. In combination therapy, the dosage and route of administration of the compound shown in Formula I and the MEK inhibitor can be determined separately.

[0033] The usage, dosage, and formulation of MEK inhibitors may be in accordance with the description in section 4, "Agents for the Prevention or Treatment of Cancer and Combination Therapies, Pharmaceutical Compositions."

[0034] Examples The present invention will be described in more detail below with reference to examples. However, the scope of the present invention is not limited to these examples. [Examples]

[0035] Orthotopic xenogram model Luciferase-expressing human pancreatic cancer cell line AsPC-1-Luc (4-hit) was orthotopically transplanted into the pancreases of seven female nude mice (BALB / cAJcl-nu / nu, n = 7 / group). Tumor luminescence intensity {photons / sec / cm2 / steradian(p / s / cm2 / sr)} was measured weekly for up to 5 weeks after administration of either a solvent alone (vehicle) or roseoflavin (RoF) and trametinib (Tr) as monotherapy or in combination (RoF+Tr).

[0036] The results are shown in Figure 1. In Figure 1, (A) RoF + Tr significantly suppressed tumor growth compared to administration of each agent alone. (B) The percentage increase in tumor luminescence intensity at 5 weeks, with the tumor luminescence intensity at the start of compound administration as the baseline, is shown in a waterfall plot. Each vertical bar represents the percentage increase in tumor luminescence intensity for one mouse. [Examples]

[0037] Allograft model (2-hit pancreatic cancer model) Eight female C57BL / 6J mice (n = 8 / group) were subcutaneously administered KPC(LSL- K RAS G12D / + ; LSL-Tr p 53 R172H / + ; Pdx1- C The re) cell line was transplanted, and the cells were administered with the solvent alone (vehicle), or with roseoflavin (RoF) and trametinib (Tr) as monotherapy or in combination (RoF + Tr).

[0038] The results are shown in Figure 2. (A) The longest diameter (L) and shortest diameter (W) of the tumor were measured twice a week until day 28, and the tumor volume (V) was calculated according to the following formula: V = (L × W^2) / 2. RoF + Tr significantly suppressed tumor growth compared to administration of each agent alone. (B) Body weight was measured five times a week until day 28. [Examples]

[0039] 0.5 μM trametinib (Tr) and RoF analogues were administered orally to 4-hit flies, respectively. The percentage of flies that emerged from all pupae in each administration group was calculated and used as the survival rate. The results are shown in Figure 3.

[0040] Administration of each analogue of RoF22, RoF23, RoF24, and RoF29 in combination with Tr significantly improved survival rates compared to the combination of RoF and Tr. Furthermore, the equilibrium dissociation constants (Kd values) of RFK and each analogue were measured using isothermal titration calorimetry. The Kd values ​​of RoF22 and RoF24 relative to RFK were lower than those of Riboflavin. RoF22 and RoF24 were shown to bind to RFK more strongly than riboflavin. [Examples]

[0041] Eight female C57BL / 6J mice in each group had KPC (LSL- K RAS G12D / + ; LSL-Tr p 53 R172H / + ; Pdx1- C The re) cell line was transplanted, and either the solvent alone (Control) or roseoflavin (RoF) or RoF22 was administered as monotherapy.

[0042] The results are shown in Figure 4. The longest (L) and shortest (W) diameters of the tumor were measured twice a week until day 21, and the tumor volume (V) was calculated according to the following formula: V = (L × W^2) / 2. RoF22 significantly suppressed tumor growth compared to RoF monotherapy. RoF22 exhibits a higher allograft growth inhibitory effect than RoF.

[0043] Furthermore, Figure 5 shows the results of weighing the subjects five times a week up to day 21. There were no significant differences in body weight among the treatment groups. RoF22 was shown not to induce weight loss. [Examples]

[0044] KPC(LSL- K RAS G12D / + ; LSL-Tr p 53 R172H / + ; Pdx1- CCell lines were transplanted, and either the solvent alone (Control) or roseoflavin (RoF) or RoF24 and trametinib (Tr) were administered as monotherapy or in combination. The longest (L) and shortest (W) diameters of the tumors were measured twice a week until day 21, and the tumor volume (V) was calculated according to the following formula: V = (L × W^2) / 2.

[0045] The results are shown in Figure 6. The combination of Tr and RoF24 significantly inhibited tumor growth compared to the combination of Tr and RoF. RoF24+Tr exhibits a higher allograft growth inhibitory effect than RoF+Tr.

[0046] Furthermore, Figure 7 shows the results of weighing the subjects five times a week up to day 21. The combination of Tr and RoF resulted in a significant weight loss compared to the control group at day 21. The combination of Tr and RoF24 did not show a significant weight difference compared to the control group at day 21. Tr+RoF24 was shown not to induce weight loss.

Claims

1. Formula I: 【Chemistry 12】 [In the formula, R １ teeth 【Chemistry 13】 Represents R ２ is -OH, -OR ２１ , or -O-CO-R ２１ (Here, R ２１ represents an alkyl group. ) The group shown in formula 11: 【Chemistry 14】 (wherein, R ３１ and R ３２ each independently represents a hydrogen atom, an alkyl group, or -CO-R ２１ .), or a group represented by -O-CON(R ４１ )(R ４２ ) (where R ４１ and R ４２ each independently represents a hydrogen atom or an alkyl group).] A therapeutic or prophylactic agent for cancer comprising the compound indicated by or a pharmaceutically acceptable salt thereof.

2. R ２ The agent according to claim 1, wherein is a hydroxyl group or an amino group.

3. The compound represented by formula I is given by the following formula: 【Chemistry 15】 The agent according to claim 1, which is represented by [the specified symbol].

4. The agent according to claim 1, wherein the cancer is a gastrointestinal cancer, respiratory cancer, urinary tract cancer, head and neck cancer, breast cancer, or hematological cancer.

5. The agent according to claim 1, wherein the cancer is pancreatic cancer, colorectal cancer, salivary gland cancer, or liver cancer.

6. The agent according to claim 1, wherein the cancer is pancreatic cancer.

7. A combination therapy agent for cancer comprising the agent according to any one of claims 1 to 3 and a MEK inhibitor.

8. The agent according to claim 7, wherein the MEK inhibitor is at least one selected from the group consisting of trametinib, binimetinib, selumetinib, mildametinib, cobimetinib, PD-334581, TAK-733, AZD8330, PD-198306, PD-98059, PD-0325901, U0126, Myrivetin, BI-847325, rifametinib, BI847325, GDC-0623, RGB-286638, Ro5126766, Isorhamnetin, CI-1040, APS-2-79, BIX02189, Hi-TOPK-032, pimasertib, PD318088, and Honokiol.

9. The agent according to claim 7, wherein the MEK inhibitor is trametinib.

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