Use of a compound in the preparation of a medicament for the prevention or treatment of a tumor

Abstract

The present application relates to a kind of compound in the application of tumor prevention or the preparation of tumor drug.The chemical formula of the compound is as follows: The compound of the present application can significantly inhibit the growth of malignant tumor, metastasis, prolong survival period;The metastasis ability of high metastatic tumor is particularly remarkable;Drug resistance large cell induced by chemotherapy drug has significant killing effect.

Description

Technical Field

[0001] This invention belongs to the field of tumor treatment technology, and specifically relates to the application of a compound in the preparation of tumor prevention or treatment drugs. Background Technology

[0002] TP53 mutations typically refer to alterations in the human TP53 gene sequence. This gene encodes a tumor suppressor protein that plays a crucial role in cell cycle control and DNA damage repair. When the TP53 gene mutates, it can lead to loss of function or abnormal function of the encoded protein, thereby affecting normal cell growth and division and increasing the risk of cancer.

[0003] TP53 gene mutations have been found in various types of cancer, including breast cancer, lung cancer, and colorectal cancer. The type and location of the mutation affect the disease to varying degrees. Some mutations can lead to complete protein inactivation, while others may only reduce protein function. Furthermore, TP53 mutations may affect an individual's response to certain cancer treatments, such as chemotherapy and radiation therapy.

[0004] Currently, there is still a need to continuously develop small molecule tumor drugs that are more active, readily available, and more affordable, especially those that can kill high-grade malignant tumors, such as those involving TP53 mutations, high metastasis, and high drug resistance. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide an application of a compound in the preparation of tumor prevention or treatment drugs. This compound can significantly inhibit the growth and metastasis of malignant tumors and prolong survival; it has a particularly significant inhibitory effect on the metastatic ability of highly metastatic tumors; and it has a significant killing effect on drug-resistant large cells induced by chemotherapy drugs.

[0006] This invention provides the application of a compound in the preparation of drugs for the prevention or treatment of tumors, the chemical formula of which is shown below: Or a pharmaceutically acceptable salt, stereoisomer, solvate, crystal form, isotope label, or prodrug thereof. The compound is a commercially novel compound, hereinafter designated as 215-3B6.

[0007] The present invention also provides the application of a pharmaceutical composition containing the aforementioned compound as an active ingredient in the preparation of drugs for the prevention or treatment of tumors.

[0008] Preferably, the pharmaceutical composition comprises an effective amount of at least one of the following: a pharmaceutically acceptable salt, solvate, crystal form, isotope-labeled derivative, or prodrug, and a pharmaceutically acceptable diluent, carrier, or excipient.

[0009] Preferably, the dosage form of the pharmaceutical composition is a tablet, capsule, injection, inhaler, or spray.

[0010] Preferably, the drug composition is administered orally, intravenously, intratumorally, or subcutaneously, and, depending on the circumstances, locally delivered to a specific tissue type (such as organs, tissues, etc.) using an appropriate method.

[0011] When intended for oral administration, the pharmaceutical composition can be formulated into oral dosage forms, such as oral solid dosage forms, like tablets, capsules, pills, granules, etc.; or oral liquid dosage forms, such as oral solutions, oral suspensions, syrups, etc. When formulated into oral dosage forms, the pharmaceutical composition may also contain suitable fillers, binders, disintegrants, lubricants, etc. When intended for parenteral administration, the pharmaceutical composition can be formulated into injectable preparations, including injection solutions, sterile powders for injection, and concentrated solutions for injection. When formulated into injectable preparations, the pharmaceutical composition can be manufactured using conventional methods in the existing pharmaceutical field, and may or may not contain excipients, or may contain suitable excipients depending on the properties of the drug.

[0012] For example, when applied topically, such as to the site of disease or to a specific organ or tissue system, such as the skin, intracranial cavity, colon, vagina, pelvis, or abdominal cavity, the formulation can be adjusted according to the specific circumstances. For instance, when used for skin cancer, the pharmaceutical composition can be formulated as a plaster, including ointments, medicated plasters, etc., with excipients added. When used for rectal or vaginal administration, the pharmaceutical composition can be formulated as suppositories, ointments, or solutions, with excipients added as needed. When used in post-operative surgical areas, the mouth, lungs, or nasopharynx, the drug can be formulated as various forms of sprays, with appropriate excipients added according to the properties of the drug. When used for pulmonary administration, the pharmaceutical composition can be formulated as an inhaler, with appropriate excipients added according to the properties of the drug.

[0013] Furthermore, the pharmaceutical composition also includes additional antitumor agents.

[0014] Furthermore, the additional antitumor agents include, but are not limited to, targeted therapies, immunotherapy drugs, chemotherapy drugs, small molecule compounds, and tumor vaccines. The combination of these additional antitumor agents includes, but is not limited to: methotrexate, capecitabine, deoxyfluorouridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, Herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, PD-L1 inhibitors, doxorubicin, PD-1 inhibitors, hydroxycamptothecin, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, traprorelin, and fluoroquinolones. A variety of antitumor agents, including tahistine, GDF15 inhibitors, leuprorelin, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, nitrogen mustard, melphalan, leukoplamide, carboplatin, cisplatin, oxaliplatin, cyclophosphamide, topotecan, camptothecin, topotecan, everolimus, sirolimus, tebuconazole, 6-mercaptopurine, 6-thioguanine, azathioprine, styraxin D, daunorubicin, doxorubicin, mitoxantrone, bleomycin, procainox, aminoglutethimide, CAR-T cells, and tumor vaccines.

[0015] Preferably, the tumor includes one or more of the following: lung cancer, liver cancer, stomach cancer, pancreatic cancer, skin cancer, head and neck cancer, myeloma, intestinal cancer, lymphoma, prostate cancer, pancreatic cancer, ovarian cancer, adrenal cancer, thyroid cancer, germ cell tumor, uterine cancer, retinoblastoma, cervical cancer, bone cancer, laryngeal cancer, urinary system tumors, oral cancer, rhabdomyosarcoma, tongue cancer, nasopharyngeal carcinoma, brain cancer, leukemia, synovial tumor, melanoma, and breast cancer.

[0016] More preferably, the tumor includes one or more of ovarian cancer, breast cancer, lung cancer, liver cancer, leukemia, colorectal cancer, melanoma, and skin cancer.

[0017] Most preferably, the tumor includes a highly metastatic tumor caused by a TP53 mutation.

[0018] The compounds described in this invention can be used in human clinical medicine and veterinary applications. These compounds can be applied to host animals, including but not limited to humans and laboratory animals such as rodents, equines, suidae, canines, felines, rabbits, bovids, and primates.

[0019] The "therapeutic effective amount" as described in this invention refers to an amount sufficient to prevent, stop, or delay disease, and to achieve, or at least partially achieve, the desired effect. Clinicians, researchers, veterinarians, and others will determine the effective amount for therapeutic use based on a variety of factors, including but not limited to the mammal species (including humans), its age, size, weight, sex, and overall health status, the severity of the cancer involved, the method of administration, timing, route of administration, and excretion rate of the drug, the bioavailability characteristics of the administered formulation, the selected dosage regimen, concurrent medication use, and whether other treatments are administered concurrently.

[0020] In some embodiments, the compound is used in combination with one or more of radiotherapy, chemotherapy, targeted therapy, immunotherapy, etc., for the prevention or treatment of tumors.

[0021] In each embodiment, unless otherwise stated, it should be understood that the formula includes and represents any and all crystalline, partially crystalline, and amorphous and / or non-crystalline forms of the compound, including partially ordered, disordered, liquid crystal, and meso-phase forms of any of the foregoing forms.

[0022] In each implementation, unless otherwise stated, it should be understood that the formula includes and represents every possible isomer, such as stereoisomers and geometric isomers, both individually and in any and all possible mixtures.

[0023] Terminology definition:

[0024] As used herein, the term "pharmaceutically acceptable salt" means a salt formed by the acidic functional group of the compound shown in this invention with a suitable inorganic or organic cation (base), and includes all forms of salt.

[0025] As used herein, the term "solvent" refers to a substance formed by the association of the compound of the present invention with a solvent molecule. The solvent may be water or an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, DMSO, etc.), including but not limited to substances that form hydrates or ethanolates.

[0026] As used herein, the term "crystal form" refers to the crystal structure of a substance. During crystallization, various factors alter the intramolecular or intermolecular bonding patterns, resulting in different arrangements of molecules or atoms in the crystal lattice, thus forming different crystal structures. The compounds of this invention can exist in one crystal structure or multiple crystal structures, i.e., they possess "polymorphism." The compounds of this invention can exist in different crystal forms.

[0027] As used herein, the term "stereoisomer" includes conformational isomers and configurational isomers, wherein the configurational isomers primarily include cis-trans isomers and optical isomers. The compounds of this invention can exist in stereoisomeric forms and therefore encompass all possible stereoisomeric forms, any combination thereof, or any mixture thereof. Examples include a single enantiomer, a single diastereomer, or a mixture of more than one. When a compound contains an olefinic double bond, unless otherwise specified, it includes cis and trans isomers, and any combination thereof.

[0028] As used herein, the term "prodrug" refers to a compound that can be converted into the present invention in a subject through reactions such as oxidation, reduction, or hydrolysis. A prodrug may or may not possess the biological activity of a compound of formula (I). Further information on the use of prodrugs can be found in Pro-drugs as Novel Delivery Systems, Vol. 14, ACSSymposium Series (T. Higuchi and W. Stella) and Bioreversible Carriers in Drug Design, Pergamon Press, 1987 (ed. E. B. Roche, American Pharmaceutical Association).

[0029] As used herein, the term “pharmaceutically acceptable carrier or excipient” means a carrier and / or excipient known in the art to be pharmacologically and / or physiologically compatible with the subject and the active ingredient.

[0030] Beneficial effects

[0031] (1) The compound (215-3B6) of the present invention can significantly inhibit the growth and metastasis of malignant tumors and prolong the survival period; it has a particularly significant inhibitory effect on the metastatic ability of highly metastatic tumors; and it has a significant killing effect on drug-resistant large cells induced by chemotherapy drugs.

[0032] (2) The compounds of this invention have a broad spectrum of inhibitory and killing effects on highly malignant tumors and are applicable to all types of tumors. Their efficacy does not depend on specific gene mutations; generally, the more metastatic the tumor, the more significant the effect. They are even more effective against highly metastatic tumors caused by TP53 mutations that do not respond well to radiotherapy and chemotherapy.

[0033] (3) The compound of the present invention has good safety in mouse experiments. After intraperitoneal inoculation (20 mg / kg) for 30 days, the weight of mice did not change significantly compared with the control group.

[0034] (4) The compounds of the present invention are commercially available, readily available, and easy to mass-produce. Attached Figure Description

[0035] Figure 1 The in vivo and in vitro antitumor effects of the compound in mouse tumor cells are shown. (A) Viability of mouse breast cancer 4T1 cells after treatment with compound 215-3B6 (10 μM) and Vehicle for 4 days (Scale bar = 50 μm). (B) Difference in growth curves between 4T1 and B16BL6 cells after treatment with compound 215-3B6 (10 μM) and Vehicle for 4 days (P < 0.0001). (C) After 4T1 cells were inoculated into mice, they were treated with Vehicle (control group) and compound 215-3B6 (20 mg / Kg); difference in survival curves between the control group and the treatment group (P < 0.0001). (D) Change in body weight of BALB / c mice after treatment with compound 215-3B6 (20 mg / Kg) for 30 days (P > 0.05). (E)BALB / c mice were treated with compound 215-3B6 (20 mg / Kg) and Vehicle for 30 minutes, respectively. The difference in body weight between the control group and the treatment group was P>0.05.

[0036] Figure 2 The in vivo and in vitro antitumor effects of the compound in human tumor cells were demonstrated. (A) Survival of human ovarian cancer OVCAR3 cells after 4 days of treatment with compound 215-3B6 (10 μM) and Vehicle, respectively (Scale bar = 50 μm). (B) Difference in tumor cell growth curves after 4 days of treatment with compound 215-3B6 (10 μM) and Vehicle, respectively (P < 0.0001). (C) Difference in tumor growth curves after intraperitoneal injection of compound 215-3B6 (20 mg / Kg) and Vehicle, respectively, following inoculation of human ovarian cancer cells OVCAR3 into nude mice to form tumors (P < 0.0001). (D) After human ovarian cancer cells OVCAR3 were inoculated into nude mice and tumors formed, the mice were treated with intraperitoneal injection of the compound 215-3B6 (20 mg / Kg) and Vehicle, respectively. After 50 days of treatment, the survival curves of the treatment group and the tumor-bearing mice were different (P<0.0001).

[0037] Figure 3 The in vitro antitumor efficacy of the compound was demonstrated in various human tumor cells. The growth curves of the tumor cells were shown to differ after 4 days of treatment with the compound 215-3B6 (10 μM) and Vehicle, respectively (P < 0.0001). Figure 4The study demonstrated the cytotoxic effect of the compound on drug-resistant large cells. (A) Drug-resistant large cells were obtained by treating 4T1 cells with the chemotherapeutic drug paclitaxel (PTX) for 5 days; the difference in the state of 4T1 tumor cells after treatment with PTX and compound 215-3B6 (10 μM) for 6 days (Scale bar = 25 μm). (B) Drug-resistant large cells were obtained by treating 4T1 cells with paclitaxel (PTX) for 5 days; the difference in the number of large cells between the PTX group and compound 215-3B6 (10 μM) after treatment with PTX and compound 215-3B6 (10 μM) for 6 days (P < 0.0001). Detailed Implementation

[0038] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0039] Example 1

[0040] Evaluation of the antitumor efficacy of the compound in mouse tumor cell lines, both in vivo and in vitro:

[0041] Cell culture medium: DMEM high glucose medium was purchased from ThermoFisher, containing 10% fetal bovine serum (Sigma); 10% penicillin-streptomycin mixed solution (ThermoFisher).

[0042] Cell lines: mouse breast cancer cell line 4T1 (TP53 mutation, extremely highly metastatic cells), mouse melanoma cell line B16BL6.

[0043] Cellular experiments: The compound was dissolved in DMSO, and 4T1 and B16BL6 cells were treated with 10 μM of the compound, with DMSO at the same final concentration as a control. After treatment with compound 215-3B6 (10 μM) for 72 hours, both 4T1 and B16BL6 cells showed significant cell death compared to the control group (P<0.0001). Figure 1 A, B).

[0044] Animal efficacy experiments: 4-6 week old BALB / c mice, weighing 19-22g, were housed in cages of 5 mice each, for a total of 4 cages. 4T1 (Luciferase-labeled) cells were seeded intramammary fat pads at a density of 5 × 10⁶ cells / mL. 5 One mouse per mouse, 20 mice inoculated.

[0045] Ten days after injection, all mice developed tumors, with a tumor diameter of approximately 0.5 cm. They were randomly divided into two groups of 10 mice each. One group received an intraperitoneal injection of the compound (20 mg / kg), while the other group received DMSO. Tumor size and mouse weight were measured every 3 days after administration. Twenty-one days after administration, two mice from each group were harvested for in vivo imaging to detect Luciferase-labeled tumor cells. The remaining eight mice, along with those receiving in vivo imaging, were observed for survival, and survival curves were plotted. The treatment lasted for 50 days. In the control group, all mice receiving DMSO died within 26 days due to tumor metastasis. Figure 1 C). After 50 days of treatment with the compound group, 7 animals (70%) still survived (P<0.0001). Figure 1 D). The results showed that the compound could inhibit tumor metastasis and significantly prolong the survival of tumor-bearing mice.

[0046] Animal safety experiments: 4-6 week old BALB / c mice, weighing 19-22g, were housed in 4 cages (5 mice / cage). They were randomly divided into two groups of 10 mice each. One group received an intraperitoneal injection of compound 215-3B6 (20 mg / kg), while the other group received an injection of DMSO. In the compound 215-3B6 treatment group, there was no significant change in mouse weight throughout the experiment (P>0.05). Figure 1 E) The mice were in good condition. There was no significant difference in body weight compared to the control group. The experimental results indicate that compound 215-3B6 has good overall safety.

[0047] Example 2

[0048] Evaluation of the antitumor efficacy of the compound in mouse tumor cell lines, both in vivo and in vitro:

[0049] Cell culture medium: DMEM high glucose medium was purchased from ThermoFisher, containing 10% fetal bovine serum (Sigma); 10% penicillin-streptomycin mixed solution (ThermoFisher).

[0050] Cell line: Human ovarian cancer cell line OVCAR3.

[0051] Cellular experiments: The compound was dissolved in DMSO, and the cells were treated with 10 μM of the compound, with DMSO at the same final concentration as a control. After treatment with compound 215-3B6 for 4 days, the growth of the cells was significantly inhibited compared with the control group (P<0.0001). Figure 2 A, B).

[0052] Animal experiments: Nude mice aged 4-6 weeks, weighing 18g-22g, were housed in separate cages (5 mice / cage), for a total of 4 cages. OCVAR1 cells were subcutaneously inoculated into the inner hind leg at a density of 5 × 10⁶ cells / mL. 5 Each mouse.

[0053] Twenty-five days after injection, all mice developed tumors, with a diameter of approximately 0.5 cm. They were randomly divided into two groups of 10 mice each. One group received an injection of compound 215-3B6 (20 mg / kg) via the pelvic cavity near the tumor inoculation site, while the other group received DMSO. Tumor size was measured every 5 days after administration. Survival time was observed, and survival curves were plotted. The treatment lasted for a total of 50 days.

[0054] Compared with the control group, the tumor growth of the compound-treated group was inhibited (P<0.0001). Figure 2 C). Compared with the control group, all animals in the compound-treated group died within 40 days after administration of DMSO. In the compound-treated group, 8 animals (80%) survived for 50 days (P<0.0001). Figure 2 D). The results showed that the compound could inhibit tumor growth and significantly prolong the survival of tumor-bearing mice.

[0055] Example 3

[0056] Evaluation of the antitumor efficacy of the compound in mouse tumor cell lines, both in vivo and in vitro:

[0057] Cell culture medium: DMEM high glucose medium was purchased from ThermoFisher, containing 10% fetal bovine serum (Sigma); 10% penicillin-streptomycin mixed solution (ThermoFisher).

[0058] Cell lines: human breast cancer cell line 231, human leukemia cell line HL60, human small cell lung cancer cell line SHP77, human glioma cell line U251, human liver cancer cell line MHCC-97H, human colon cancer cell line HCT116p53- / -(p53N), and human prostate cancer cell line PC3.

[0059] Cell experiments: Compound 215-3B6 was dissolved in DMSO, and the cells were treated with 15 μM of the compound, with DMSO at the same final concentration used as a control. After 4 days of treatment with compound 215-3B6, cell growth was significantly inhibited compared to the control group (P < 0.0001). Figure 3 Experimental results show that compound 215-3B6 has a universal inhibitory effect on tumor cells.

[0060] Example 4

[0061] The compound kills paclitaxel-induced drug-resistant tumor giant cells:

[0062] Cell culture medium: DMEM high glucose medium was purchased from ThermoFisher, containing 10% fetal bovine serum (Sigma); 10% penicillin-streptomycin mixed solution (ThermoFisher).

[0063] Cell line: Mouse breast cancer cell line 4T1 (TP53 mutation, extremely highly metastatic cells).

[0064] Cellular experiments: The compound (215-3B6) and the chemotherapeutic drug paclitaxel (PTX) were dissolved in DMSO. 4T1 cells were treated with 10 μM paclitaxel (PTX) for 5 days to obtain 4T1 cells containing a large number of drug-resistant cells. The treated cells were divided into two groups: one group was still treated with 10 μM paclitaxel (PTX), and the other group was treated with 10 μM of the compound 215-3B6.

[0065] After 72 hours of treatment with the compound, compared with the control group (PTX treatment), the 4T1-resistant tumor giant cells in the compound-treated group showed significant death. Figure 4 A), the quantity decreased drastically (P<0.0001) Figure 4 B). Experimental results show that the compound (215-3B6) has a killing effect on drug-resistant tumor giant cells.

Claims

1. The application of a compound in the preparation of drugs for the prevention or treatment of tumors, characterized in that: The chemical formula of the compound is shown below: Or a pharmaceutically acceptable salt, stereoisomer, solvate, crystal form, isotope label, or prodrug.

2. The use of a pharmaceutical composition containing the compound as described in claim 1 as an active ingredient in the preparation of a drug for the prevention or treatment of tumors.

3. The application according to claim 2, characterized in that, The pharmaceutical composition comprises an effective amount of at least one of the following: a compound or a pharmaceutically acceptable salt, solvate, crystal form, isotope-labeled derivative, or prodrug, and a pharmaceutically acceptable diluent, carrier, or excipient.

4. The application according to claim 2, characterized in that, The dosage form of the pharmaceutical composition is tablets, capsules, injections, inhalers, or sprays.

5. The application according to claim 2, characterized in that, The drug composition can be administered orally, intravenously, intratumorally, or subcutaneously.

6. The application according to claim 2, characterized in that, The pharmaceutical composition also includes additional antitumor agents.

7. The application according to claim 6, characterized in that, The additional antitumor agents include, but are not limited to, one or more of targeted drugs, immunotherapy drugs, chemotherapy drugs, small molecule compounds, and tumor vaccines.

8. The application according to claim 2, characterized in that: The tumors include one or more of the following: lung cancer, liver cancer, stomach cancer, pancreatic cancer, skin cancer, head and neck cancer, myeloma, intestinal cancer, lymphoma, prostate cancer, pancreatic cancer, ovarian cancer, adrenal cancer, thyroid cancer, germ cell tumor, uterine cancer, retinoblastoma, cervical cancer, bone cancer, laryngeal cancer, urinary system tumors, oral cancer, rhabdomyosarcoma, tongue cancer, nasopharyngeal carcinoma, brain cancer, leukemia, synovial tumor, melanoma, and breast cancer.

9. The application according to claim 8, characterized in that: The tumors include one or more of the following: ovarian cancer, breast cancer, lung cancer, liver cancer, leukemia, colorectal cancer, melanoma, and skin cancer.

10. The application according to claim 8, characterized in that: The tumors include highly metastatic tumors caused by TP53 mutations.

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