Dual-target drug or pharmaceutical composition for preventing, alleviating, or treating cancer, treatment method, and use

Abstract

The present application relates to a drug or pharmaceutical composition for preventing, alleviating, or treating cancer and a use thereof, as well as a method for preventing, alleviating, or treating cancer. The drug or pharmaceutical composition of the present application is capable of interfering with ataxia-telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR) activity, causing inactivation or reduced activity of ATR and mTOR. The present application provides a novel dual-target strategy for the treatment of cancer.

Description

Dual-target drugs or drug combinations for the prevention, relief or treatment of cancer, treatment methods and uses. Technical Field

[0001] This application belongs to the field of biomedical technology, specifically relating to a drug or pharmaceutical composition for the prevention, relief or treatment of cancer, a method for the prevention, relief or treatment of cancer, and a pharmaceutical use. More specifically, this application relates to a drug or pharmaceutical composition capable of intervening in the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR). Background Technology

[0002] As one of the leading causes of death worldwide, cancer seriously threatens human health. In 2022, the number of newly diagnosed cancer cases globally reached 19.96 million, with China accounting for 24.1%; in terms of cancer deaths, there were approximately 9.7 million cases globally, with China accounting for 26.5%. According to WHO statistics, an average of 7 million people worldwide die from malignant tumors each year; the number of cancer deaths worldwide will continue to rise, and is projected to exceed 13.1 million by 2030. In terms of tumor types, lung cancer, breast cancer (in women), and colorectal cancer are the three most frequently diagnosed cases globally; while in terms of the number of deaths, lung cancer, colon cancer, and liver cancer rank among the top three. In China, in addition to the aforementioned tumor types, esophageal cancer ranks among the top ten cancers in terms of both incidence and mortality. Therefore, cancer has become the most concentrated area for new drug development globally.

[0003] Surgery, radiotherapy, and chemotherapy have a long history of use as primary treatments for cancer. Both surgery and radiotherapy are localized treatments. Surgery is most effective for benign or early-stage malignant solid tumors that have not yet spread or metastasized, but its effectiveness is very limited for non-solid tumors such as those in hematologic malignancies, as well as for tumors that have spread or metastasized. It is difficult to completely eliminate lesions, and there is a risk of recurrence and organ dysfunction. Radiotherapy can be used as a primary treatment for tumors or as adjuvant therapy before or after surgery. Its limitations lie mainly in the varying sensitivity of different tumors to radiotherapy and significant toxic side effects. Chemotherapy, as a systemic treatment, can kill both solid and non-solid tumors, as well as metastases. However, chemotherapy tolerance is the most challenging issue, often manifesting as systemic side effects; moreover, different cancer types have varying sensitivities to chemotherapy. Against this backdrop, the emergence of targeted cancer drugs aims to target tumors with precision medicine, developing systemic therapies that are selective in their targets, specific in their mechanisms, and well-tolerated. Since the 1960s, with the development of scientific research and technology, a continuous stream of targeted cancer therapies based on various technologies, including small molecule chemical drugs, antibodies and antibody-drug conjugates, and cell and gene therapy, has emerged globally. Among these, small molecule chemical drugs remain the primary form of targeted cancer drug development. Their development strategy involves designing small molecules to intervene in the activity of protein molecules that exhibit mutations and functional abnormalities in tumor cells, driving their unlimited growth and metastasis, thereby inhibiting or even killing tumor cells. However, in clinical application, existing drugs developed targeting single genes or single targets suffer from insufficient efficacy, drug resistance, relapse, and toxicity. Furthermore, existing cancer treatments targeting single genes or single targets lack a sufficient safety window. Therefore, finding more rational drug development strategies and cancer treatment strategies with a larger safety window is urgently needed. Summary of the Invention

[0004] To address at least one of the aforementioned problems, this application provides a dual-target strategy for the prevention, mitigation, or treatment of cancer, offering a new therapeutic approach for treating cancer, particularly for cancers that have long plagued humanity, such as melanoma, esophageal cancer, or colon cancer.

[0005] The first aspect of this application provides a medicament or pharmaceutical composition for the prevention, relief or treatment of cancer, characterized in that the medicament or pharmaceutical composition is capable of interfering with the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR), thereby inactivating or reducing the activity of ATR and mTOR.

[0006] A second aspect of this application provides a method for preventing, alleviating, or treating cancer, characterized in that the method comprises administering to a subject in need a therapeutically effective dose of a drug or pharmaceutical composition capable of interfering with the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR), said drug or pharmaceutical composition inactivating or reducing the activity of ATR and mTOR.

[0007] A third aspect of this application provides the use of a medicament or pharmaceutical composition capable of intervening in the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR) in the preparation of a medicament or pharmaceutical composition for the prevention, relief or treatment of cancer, wherein the medicament or pharmaceutical composition inactivates or reduces the activity of ATR and mTOR.

[0008] The dual-target strategy provided in this application demonstrates the synergistic effect of targeting both ATR and mTOR, exhibiting a stronger tumor-suppressing effect compared to single-target ATR or mTOR strategies. Since single-target drugs targeting ATR or mTOR, especially chemotherapy drugs, may have certain toxic side effects, the dual-target strategy of ATR and mTOR may increase these side effects and reduce safety. Therefore, currently, there are no single or combined drugs or drug compositions using the dual-target strategy of ATR and mTOR for tumor treatment or prevention in clinical trials or clinical applications. This application finds that the dual-target strategy of ATR and mTOR achieves synergistic effects while also having good safety, providing a new approach to tumor treatment.

[0009] It should be understood that the description in this section is not intended to identify key or essential features of the embodiments of this application, nor is it intended to limit the scope of this application. Other features of this application will become readily apparent from the following description. Attached Figure Description

[0010] The accompanying drawings exemplify embodiments and form part of the specification, serving together with the textual description to explain exemplary implementations of the embodiments. The illustrated embodiments are for illustrative purposes only and do not limit the scope of the claims. Throughout the drawings, the same reference numerals refer to similar but not necessarily identical elements.

[0011] Figure 1 shows the growth curves of tumor volume in each group of mice in the human esophageal cancer KYSE410 subcutaneous xenograft model in Example 3.

[0012] Figure 2 shows the weight change curves of mice in each group of the human esophageal cancer KYSE410 subcutaneous xenograft model as of treatment time in Example 3.

[0013] Figure 3 shows the growth curves of tumor volume in each group of mice in the NCI-H460 human large cell lung cancer subcutaneous xenograft model in Example 4.

[0014] Figure 4 shows the weight change curves of mice in each group of the NCI-H460 human large cell lung cancer subcutaneous xenograft model as of treatment time in Example 4.

[0015] Figure 5. Dose-response matrix and synergistic score of ATR inhibitor M4344 combined with mTOR inhibitor AZD2014 in NCI-H460 cells.

[0016] Figure 6. Dose-response matrix and synergistic score of ATR inhibitor M4344 combined with mTOR inhibitor CC223 in NCI-H460 cells.

[0017] Figure 7. Dose-response matrix and synergistic score of ATR inhibitor AZD6738 combined with mTOR inhibitor AZD2014 in NCI-H460 cells.

[0018] Figure 8. Dose-response matrix and synergistic score of ATR inhibitor AZD6738 combined with mTOR inhibitor CC223 in NCI-H460 cells.

[0019] Figure 9. Dose-response matrix and synergistic score of ATR inhibitor M4344 combined with mTOR inhibitor AZD2014 in B16-F10 cells.

[0020] Figure 10. Dose-response matrix and synergistic score of ATR inhibitor M4344 combined with mTOR inhibitor CC223 in B16-F10 cells.

[0021] Figure 11. Dose-response matrix and synergistic score of ATR inhibitor AZD6738 combined with mTOR inhibitor AZD2014 in B16-F10 cells.

[0022] Figure 12. Dose-response matrix and synergistic score of ATR inhibitor AZD6738 combined with mTOR inhibitor CC223 in B16-F10 cells. Detailed Implementation

[0023] To make the above-mentioned objectives, features and advantages of this application more apparent and understandable, the specific embodiments of this application are described in detail below.

[0024] Unless otherwise stated, all figures used in this specification and claims to represent content, concentration, proportion, weight, particle size, percentage, technical effect, etc., should in any instance be understood to be modified by the terms “about” or “approximately”. Therefore, unless indicated to the contrary, the numerical parameters listed in the following specification and appended claims are approximate values. Unless otherwise stated, the terminology used herein has its common meaning as understood by one of ordinary skill in the art. It may vary for those skilled in the art depending on the desired properties and effects sought through this application, and each numerical parameter should be interpreted according to the number of significant figures and conventional rounding methods or as understood by one of ordinary skill in the art.

[0025] While the numerical ranges and parameters described in this application are approximate, the values ​​presented in the specific embodiments are provided as precisely as possible. However, any numerical value will inherently contain some errors, which are necessarily caused by the standard deviation found in its corresponding test measurements. Each numerical range given in this specification will include every narrower numerical range falling within that wider range, as if these narrower numerical ranges were explicitly stated herein.

[0026] Unless otherwise stated or contradicted by the context, the terms or expressions used herein should be read in the context of the document and as understood by one of ordinary skill in the art. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0027] When used in this document, the expression “A and / or B” includes three cases: (1) A; (2) B; and (3) A and B. The expression “A, B and / or C” includes seven cases: (1) A; (2) B; (3) C; (4) A and B; (5) A and C; (6) B and C; and (7) A, B and C. The meanings of similar expressions can be deduced by analogy.

[0028] Generally, the nomenclature used herein and the experimental procedures in organic chemistry, medicinal chemistry, and biology described herein are well-known and commonly used in the field. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. Where multiple definitions exist for terms used herein, the definitions in this section shall prevail unless otherwise stated.

[0029] Drugs or drug compositions

[0030] In a first aspect, this application provides a drug or pharmaceutical composition for the prevention, relief or treatment of cancer, characterized in that the drug or pharmaceutical composition is capable of interfering with the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR), thereby inactivating or reducing the activity of ATR and mTOR.

[0031] When used in this document, the term “prevention” and its synonyms refer to delaying the onset of a particular disease, condition and / or condition or symptoms associated with such disease, condition and / or condition, or reducing the risk of acquiring such disease, condition and / or condition.

[0032] As used herein, the terms “relief” and “treatment” and their synonyms refer to improvement of a disease, symptom, and / or condition. “Relief” and “treatment” can be an improvement in at least one measurable physical parameter, which is not necessarily identifiable by the subject. “Relief” and “treatment” can also be physical (e.g., stabilizing identifiable symptoms), physiological (e.g., stabilizing physical parameters), or both, suppression of the progression of a disease, symptom, and / or condition. “Relief” and “treatment” can also be the slowing of or reversal of the progression of a disease, symptom, and / or condition.

[0033] As used herein, the term "drug" refers to at least one active ingredient that can affect the physiological functions of organs and / or cellular metabolic activities. When a drug comprises two or more active ingredients, these two or more active ingredients may be included in the same formulation or in different formulations. When two or more active ingredients are included in different formulations, the different formulations may be administered simultaneously or separately at appropriate intervals, for example, administering the first formulation first, followed by the second formulation after 5 minutes, 10 minutes, 6 hours, or 1 day, etc.

[0034] ATR is a major regulator of DNA damage or replication stress, playing a protective role in the cell nucleus and safeguarding genome stability. mTOR is a key signaling molecule for cellular functions such as synthesis and catabolism. The mTOR signaling pathway is closely related to nutrition, energy status, and growth factors. However, the effects of simultaneously reducing or inactivating the activities of ATR and mTOR on the occurrence and development of esophageal cancer, colon cancer, and other tumors are currently unclear and have not been reported in the literature. This application finds that simultaneously reducing the activities of ATR and mTOR has a synergistic effect on the treatment of cancers such as esophageal and colon cancer, providing a new approach to cancer treatment.

[0035] In some embodiments, the cancer includes at least one of esophageal cancer, gastroesophageal junction cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, melanoma, lung cancer, thyroid cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, brain cancer, neuroendocrine tumor, kidney cancer, urothelial carcinoma, bladder cancer, prostate cancer, liver cancer, leukemia, lymphoma, multiple myeloma, sarcoma, or glioma.

[0036] In some embodiments, the esophageal cancer includes at least one of esophageal squamous cell carcinoma or esophageal adenocarcinoma.

[0037] In some embodiments, and / or, the melanoma includes at least one of acral melanoma, mucosal melanoma, or cutaneous melanoma.

[0038] In some embodiments, the lung cancer includes at least one of non-small cell lung cancer, small cell lung cancer, or large cell lung cancer.

[0039] In some embodiments, the breast cancer includes at least one of Her2-positive breast cancer or Her2-negative breast cancer.

[0040] In some embodiments, the brain tumor includes at least one of glioma, glioblastoma, or astrocytoma.

[0041] In some embodiments, the neuroendocrine tumor includes at least one of a pulmonary neuroendocrine tumor, a gastric neuroendocrine tumor, an enteric neuroendocrine tumor, or a pancreatic neuroendocrine tumor.

[0042] In some embodiments, the cancer is selected from esophageal cancer. In some embodiments, the cancer is selected from esophageal squamous cell carcinoma.

[0043] In some implementations, the cancer is selected from colon cancer.

[0044] In some embodiments, the activity of the intervention ATR includes at least one of the following:

[0045] Inhibit the interaction between ATR and ATRIP;

[0046] This causes the complex of ATR and ATRIP to dissociate;

[0047] Inhibit the interaction between ATR and TopBP1;

[0048] This causes the complex of ATR and TopBP1 to dissociate;

[0049] Inhibit the interaction between ATR and ETAA1;

[0050] This causes the complex of ATR and ETAA1 to dissociate;

[0051] Degrade at least one of ATR, ATRIP, TopBP1, ETAA1, ATR and ATRIP complex, ATR and TopBP1 complex, or ATR and ETAA1 complex.

[0052] Inhibit the activity of ATR protein;

[0053] Inhibit ATR autophosphorylation;

[0054] Inhibit ATR phosphorylation of at least one of CHK1, p53, SMARCAL1, WRN, or FANCI;

[0055] Repressing the transcription and / or translation of atr genes; and / or

[0056] This can cause deletion and / or mutation of the atr gene.

[0057] ATR is a serine-threonine kinase in the phosphatidylinositol 3-kinase (PIKK) family, and its activation and regulation are a complex, multi-step process. ATR binds to its regulatory partner protein, ATR-ATRIP, forming an ATR-ATRIP complex. This complex is enriched at DNA damage sites by DNA damage-induced replication extension protein A (RPA)-coated single-stranded DNA (RPA-ssDNA), thus exerting its bioregulatory role. Further, after being enriched at DNA damage sites, ATR undergoes autophosphorylation at its T1989 residue. Topoisomerase 2-binding protein 1 (TopBP1) binds to these autophosphorylated residues, further activating ATR. Ewing tumor-associated antigen 1 (ETAA1), also a regulatory partner protein of ATR, can bind to ATR and promote its activation. Once activated, ATR can exert its biological activity by phosphorylating various downstream substrates, such as checkpoint kinase 1 (CHK1), p53, SMARCAL1, WRN, or FANCI. When at least one of the above-mentioned biological processes is inhibited, or at least one related complex is dissociated or degraded, the activity of ATR may be interfered with, which may lead to the inactivation or reduction of ATR activity.

[0058] In some embodiments, the intervention on mTOR activity includes at least one of the following:

[0059] Inhibit mTOR activity;

[0060] Inhibits the activity of at least one of mTORC1 or mTORC2;

[0061] Inhibit the generation of at least one of mTORC1 or mTORC2;

[0062] To cause at least one of mTORC1 or mTORC2 to dissociate;

[0063] Degrade at least one of mTOR, mTORC1, or mTORC2;

[0064] Repressing the transcription and / or translation of the mtor gene; and / or

[0065] This can cause deletion and / or mutation of the mtor gene.

[0066] mTOR can recruit related proteins and active factors to assemble into two types of mTOR complexes, exerting biological regulatory functions. For example, mTOR binds to RAPTOR, mLST8, and two negative regulators, PRAS40 and Deptor, to form mTOR complex 1 (mTORC1), while mTOR binds to Protor, Rictor, Deptor, mSlN1, and mLST8 to form mTOR complex 2 (mTORC2). mTORC1 or mTORC2 can phosphorylate S6, AKT, etc., activating the biological activity of substrates and playing a regulatory role in biological processes. mTOR mainly participates in the regulation of four signaling pathways, including the PI3K / AKT / mTORC2 pathway, the Ras / MAPK / mTORC1 pathway, the Wnt / GSK3β / mTORC1 pathway, and the ULK-ATG13-FIP200 pathway.

[0067] In some embodiments, the drug or drug composition includes at least one of protein, polypeptide, nucleic acid, polynucleotide, polynucleotide, antibody or derivative thereof, organic synthetic compound, inorganic compound and / or natural product.

[0068] As used herein, the term "peptide" refers to a polymer of amino acids of any length.

[0069] When used in this document, the terms “nucleic acid” and “polynucleotide” are used interchangeably and refer to a polymer of nucleotides of any length, including deoxyribonucleotides, ribonucleotides, combinations thereof, and analogues.

[0070] As used herein, the term "antibody" refers to a specific immunoglobulin or antigen-binding fragment thereof targeting an antigen site. In this application, "antibody" refers to an antibody that specifically binds to the ATR and / or mTOR of this application and can be manufactured according to methods known in the art. Antibodies can take the form of polyclonal or monoclonal antibodies, antibody fragments (such as Fab, Fab', F(ab')2, and Fv fragments), single-chain Fv (scFv) antibodies, multispecific antibodies (such as bispecific antibodies), monospecific antibodies, monovalent antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins containing an antigen-binding site, and any other modified immunoglobulin molecule containing an antigen-binding site, provided that the antibody exhibits the desired biological binding activity.

[0071] Those skilled in the art can select appropriate types of drugs or reagents as needed to intervene in the activity of ATR and mTOR, thereby inactivating or reducing their activity. For example, when intervention in ATR and mTOR is required, common methods for inhibiting protein expression levels or activity can be used, including but not limited to antibodies, small molecule inhibitors, and chimeras targeting protein degradation. If intervention in the encoding nucleic acids of ATR and mTOR is required, common methods for inhibiting nucleic acid expression levels or gene regulation can be used, including but not limited to gene knockout, antisense nucleic acids, ribozymes, or interfering RNA technology. Those skilled in the art can prepare the above-mentioned types of drugs or reagents using methods known in the art to intervene in the activity of ATR and mTOR.

[0072] In some embodiments, the drug or drug composition includes compounds that inhibit ATR activity and mTOR activity.

[0073] In some embodiments, the drug or drug composition includes an ATR inhibitor and an mTOR inhibitor.

[0074] In some embodiments, the ATR inhibitor comprises at least one of AZD6738, M-4344, M6620, BAY-1895344, RP-3500, M1774, ART0380, SKLB-197, AD1058, or a tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt, or a solvate of a pharmaceutically acceptable salt thereof; and / or

[0075] The mTOR inhibitors include sirolimus, zotalimus, umirolimus, tesimolimus, gidalixetine, Paxalisib, SR-0379, Dactolisib, Oleanin, Olcorolimus, Everolimus, RM-006, desfolimus, SF-1126, KB02-SLF, LY-3023414, TAK-228, AZD2014, CC223, CC-115, RTB-101, Voxtalisib, ME-344, SCC-31, CC-214-2, DHW-208, FD-274, GNE-477, GNE-555, Panulisib, PF-04691503, Piplartine, PP30, PQR514, Torin2, Torkinib, X-387, PQR- 530, Torin1, AZD8055, KU-0063794, BGT-226, Bimiralisib, GDC-0349, Omilise, OSI-027, PF-0469 1502, PKI-179, XL-765, Apitolisib, DS-7423, GNE-493, MKC-1, P-2281, Palomids, PI-103, PWT- 33597, SN-32976, WYE-132, XL-388, PP242, PP30, XL388, WYE-354, WAY-600, WYE-687, AP23573, MLN0128 or at least one of the following: tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt or solvate of a pharmaceutically acceptable salt thereof.

[0076] As used herein, the term "tautomer" refers to two isomers containing heteroatoms (such as nitrogen, oxygen, or sulfur atoms) whose structural differences lie only in the migration of protons and the corresponding double bonds, and which coexist in an equilibrium system, transforming into each other at a fairly high rate. In some embodiments, keto-enol tautomerism is a more common "tautomerism" phenomenon.

[0077] When used in this article, the term "stereoisomer" refers to an isomer resulting from the different spatial arrangements of atoms in a molecule.

[0078] When used in this document, the term "enantiomer" refers to a stereoisomer that is a real object and its mirror image but cannot be superimposed.

[0079] When used in this document, the term “diastereomer” refers to a stereoisomer that has two or more chiral centers and is not a mirror image of the other molecules.

[0080] When used in this document, the term "racemate" refers to an equimolar mixture of an optically active chiral molecule and its enantiomer.

[0081] As used herein, the term "solvent" refers to the association of one or more solvent molecules with an organic synthetic compound, inorganic compound, and / or natural product or its salt thereof. Solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, ethyl acetate, or acetic acid.

[0082] In some embodiments, the solvate or a pharmaceutically acceptable salt thereof comprises at least one of a monohydrate, a dihydrate, a trihydrate, a monomethanol, a dimethanol, a monoacetonitrile, a diacetonitrile, a monoacetone, a diacetone, a hemifumarate monohydrate, a fumarate dihydrate, or a fumarate-ethanol.

[0083] As used herein, the term "isotope derivative" refers to a structure having the chemical formula described by the organic synthetic compound, inorganic compound, and / or natural product given in this application, wherein one or more atoms are replaced by isotopic atoms having a selected atomic mass or mass number. Isotopes that can be incorporated into the organic synthetic compound, inorganic compound, and / or natural product of this application include, for example, isotopes of hydrogen. In some specific embodiments, incorporation of certain isotopes, particularly deuterium (i.e., 2H or D), can provide certain therapeutic advantages by providing greater metabolic stability, such as increasing the in vivo half-life or reducing dose requirements or improving the therapeutic index or tolerability.

[0084] In some embodiments, the isotopes that may be incorporated into the organic synthetic compounds, inorganic compounds, and / or natural products of this application include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, chlorine, fluorine, or iodine, such as 3H, 11C, 13C, 14C, 15N, 18F, 35S, 36Cl, 123I, 124I, or 125I, respectively. In some embodiments, this application includes compounds incorporated with one or more of the above isotopes, for example, including portions incorporated with the radioactive isotopes 3H and / or 14C, or portions incorporated with the non-radioactive isotopes 2H and / or 13C.

[0085] In some embodiments, the organic synthetic compounds, inorganic compounds and / or natural products of this application may exist in free form or in the form of their salts.

[0086] As used herein, the term "pharmaceutically acceptable salt" means a salt that retains the desired biological activity of the organic synthetic compounds, inorganic compounds, and / or natural products of this application and exhibits lower undesirable toxicological effects. These pharmaceutically acceptable salts may be prepared in situ during the final separation and purification of the compound, or by reacting the purified compound, in its free acid or free base form, separately with a suitable base or acid.

[0087] In some embodiments, an inorganic acid and an organic acid can be used to form a pharmaceutically acceptable acid addition salt. The inorganic acid includes hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, or phosphoric acid, etc. The organic acid includes acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, or sulfosalicylic acid, etc.

[0088] In some embodiments, a pharmaceutically acceptable base addition salt can be formed using an inorganic base and an organic base. The inorganic base includes, for example, ammonium salts and metals from columns I to XII of the periodic table. The organic base includes, for example, primary, secondary, and tertiary amines; substituted amines (including naturally occurring substituted amines); cyclic amines; basic ion exchange resins, etc. In some specific embodiments, the organic base includes isopropylamine, benzylamine, choline salts, diethanolamine, diethylamine, lysine, meglumine, piperazine, or tromethamine, etc.

[0089] In some embodiments, the pharmaceutically acceptable salt includes at least one of hydrochloride, sulfate, phosphate, oxalate, maleate, methanesulfonate, succinate, citrate, fumarate, glucuronide, formate, acetate, or succinate.

[0090] When used in this article, AZD6738 has the following chemical structural formula:

[0091] When used in this document, M-4344 has the following chemical structural formula:

[0092] When used in this document, M6620 has the following chemical structural formula:

[0093] When used in this article, BAY-1895344 has the following chemical structural formula:

[0094] When used in this article, RP-3500 has the following chemical structural formula:

[0095] When used in this article, M1774 has the following chemical structural formula:

[0096] When used in this article, ART0380 has the following chemical structural formula:

[0097] When used in this article, SKLB-197 has the following chemical structural formula:

[0098] When used in this article, AD1058 has the following chemical structural formula:

[0099] When used in this article, sirolimus has the following chemical structural formula:

[0100] When used in this article, zotamoxetine has the following chemical structural formula:

[0101] In this paper, Umirolimus is referred to by the formula, which has the following chemical structural formula.

[0102] When used in this article, tesimolimus has the following chemical structural formula:

[0103] When used in this article, Gidarice has the following chemical structural formula:

[0104] When used in this article, Paxalisib has the following chemical structural formula:

[0105] When used in this article, Dactolisib has the following chemical structural formula:

[0106] When used in this article, Oleandrin has the following chemical structural formula:

[0107] When used in this article, Everolimus has the following chemical structural formula:

[0108] When used in this article, RM-006 has the following chemical structural formula:

[0109] When used in this article, desphosmoxine has the following chemical structural formula:

[0110] When used in this article, SF-1126 has the following chemical structural formula:

[0111] When used in this article, KB02-SLF has the following chemical structural formula:

[0112] When used in this article, LY-3023414 has the following chemical structural formula:

[0113] When used in this article, TAK-228 has the following chemical structural formula:

[0114] When used in this article, AZD2014 has the following chemical structural formula:

[0115] When used in this article, CC223 has the following chemical structural formula:

[0116] When used in this article, CC-115 has the following chemical structural formula:

[0117] When used in this article, RTB-101 has the following chemical structural formula:

[0118] When used in this article, Voxtalisib has the following chemical structural formula:

[0119] When used in this article, ME-344 has the following chemical structural formula:

[0120] When used in this article, SCC-31 has the following chemical structural formula:

[0121] When used in this article, CC-214-2 has the following chemical structural formula:

[0122] When used in this article, DHW-208 has the following chemical structural formula:

[0123] When used in this article, FD-274 has the following chemical structural formula:

[0124] When used in this article, GNE-477 has the following chemical structural formula:

[0125] When used in this article, GNE-555 has the following chemical structural formula:

[0126] When used in this article, Panulisib has the following chemical structural formula:

[0127] When used in this article, PF-04691503 has the following chemical structural formula:

[0128] When used in this article, Piplartine has the following chemical structural formula:

[0129] When used in this article, PP30 has the following chemical structural formula:

[0130] When used in this article, PQR514 has the following chemical structural formula:

[0131] When used in this article, Torin2 has the following chemical structural formula:

[0132] When used in this article, Torkinib has the following chemical structural formula:

[0133] When used in this article, X-387 has the following chemical structural formula:

[0134] When used in this article, PQR-530 has the following chemical structural formula:

[0135] When used in this article, Torin1 has the following chemical structural formula:

[0136] When used in this document, AZD8055 has the following chemical structural formula:

[0137] When used in this article, KU-0063794 has the following chemical structural formula:

[0138] When used in this article, BGT-226 has the following chemical structural formula:

[0139] When used in this article, Bimiralisib has the following chemical structural formula:

[0140] When used in this article, GDC-0349 has the following chemical structural formula:

[0141] When used in this article, Omilise has the following chemical structural formula:

[0142] When used in this article, OSI-027 has the following chemical structural formula:

[0143] When used in this document, PF-04691502 has the following chemical structural formula:

[0144] When used in this article, PKI-179 has the following chemical structural formula:

[0145] When used in this article, XL-765 has the following chemical structural formula:

[0146] When used in this article, Apitolisib has the following chemical structural formula:

[0147] When used in this article, DS-7423 has the following chemical structural formula:

[0148] When used in this article, GNE-493 has the following chemical structural formula:

[0149] When used in this article, MKC-1 has the following chemical structural formula:

[0150] When used in this article, P-2281 has the following chemical structural formula:

[0151] When used in this article, Palomidas have the following chemical structural formula:

[0152] When used in this article, PI-103 has the following chemical structural formula:

[0153] When used in this article, PWT-33597 has the following chemical structural formula:

[0154] When used in this article, SN-32976 has the following chemical structural formula:

[0155] When used in this article, WYE-132 has the following chemical structural formula:

[0156] When used in this article, XL-388 has the following chemical structural formula:

[0157] When used in this article, PP242 has the following chemical structural formula:

[0158] When used in this article, PP30 has the following chemical structural formula:

[0159] When used in this article, XL388 has the following chemical structural formula:

[0160] When used in this article, WYE-354 has the following chemical structural formula:

[0161] When used in this article, WAY-600 has the following chemical structural formula:

[0162] When used in this article, WYE-687 has the following chemical structural formula:

[0163] When used in this article, AP23573 has the following chemical structural formula:

[0164] When used in this article, MLN0128 has the following chemical structural formula:

[0165] In some embodiments, the ATR inhibitor is selected from at least one of AZD6738, M-4344, or their tautomers, stereoisomers, enantiomers, diastereomers, racemic mixtures, solvates, isotope derivatives, pharmaceutically acceptable salts, or solvates of pharmaceutically acceptable salts thereof; and / or

[0166] The mTOR inhibitors include at least one of AZD2014, CC223 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

[0167] In some embodiments, the ATR inhibitor is selected from M-4344 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof; and the mTOR inhibitor comprises AZD2014 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

[0168] In some embodiments, the dosage form of the drug or drug composition includes at least one of the following: solution, powder, granule, tablet, sugar-coated, capsule, granule, suspension, syrup, drops, and sublingual tablet.

[0169] Those skilled in the art can select appropriate dosage forms of drugs or drug compositions based on actual usage needs.

[0170] In some embodiments, the pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0171] As used herein, the term "pharmaceuticalally acceptable excipient" refers to a pharmaceutically acceptable raw material, component, or carrier that imparts a form or consistency to the pharmaceutical composition. When mixed, each excipient is compatible with the other components of the pharmaceutical composition, thereby avoiding a significant reduction in the potency of the drug when administered to a subject and avoiding interactions between pharmaceutically unacceptable drug components.

[0172] In some embodiments, the excipients include, but are not limited to, diluents, fillers, binders, disintegrants, lubricants, flow aids, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, taste masking agents, colorants, anti-caking agents, humectants, chelating agents, plasticizers, thickeners, antioxidants, preservatives, stabilizers, surfactants, or buffers. Those skilled in the art may selectively use the above-mentioned pharmaceutically acceptable excipients depending on the dosage form of the pharmaceutical composition.

[0173] In some embodiments, the excipients include at least one of solubilizers, disintegrants, wetting agents, stabilizers, thickeners, diluents, buffers, and flavoring agents.

[0174] Methods to prevent, alleviate or treat cancer

[0175] Secondly, this application provides a method for preventing, alleviating, or treating cancer, the method comprising administering to a subject in need a therapeutically effective dose of a drug or pharmaceutical composition capable of intervening in the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR), said drug or pharmaceutical composition inactivating or reducing the activity of ATR and mTOR.

[0176] As used herein, the term "subject" refers to a primate (e.g., a human), a dog, a rabbit, a guinea pig, a pig, a rat, and a mouse. In some embodiments, the subject is a primate. In some specific embodiments, the subject is a human.

[0177] When used in this article, a subject is considered "needing" the treatment if the subject benefits from it biologically, medically, or in terms of quality of life.

[0178] As used herein, the term "therapeutic effective dose" refers to an amount that, compared to a corresponding subject who did not receive that amount, results in a benefit or treatment of a disease, but is sufficiently low within the range of reasonable medical judgment to avoid serious side effects. The therapeutic effective dose of a drug or pharmaceutical composition will vary depending on factors such as the specific drug chosen (e.g., taking into account the drug's potency, efficacy, and half-life); the chosen route of administration; the disease being treated; the severity of the disease being treated; the age, body type, weight, and physical condition of the patient being treated; the patient's medical history; the duration of treatment; the nature of concurrent treatments; and the desired therapeutic effect, but can still be determined by those skilled in the art in a conventional manner.

[0179] In some embodiments, the cancer includes at least one of the following: esophageal cancer, gastroesophageal junction cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, melanoma, lung cancer, thyroid cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, brain cancer (including glioma, glioblastoma, astrocytoma, etc.), neuroendocrine tumor, kidney cancer, urothelial carcinoma, bladder cancer, prostate cancer, liver cancer, leukemia, lymphoma, multiple myeloma, sarcoma, or glioma.

[0180] In some embodiments, the esophageal cancer includes at least one of esophageal squamous cell carcinoma or esophageal adenocarcinoma; and / or, the melanoma includes at least one of acral melanoma, mucosal melanoma, or cutaneous melanoma; and / or, the lung cancer includes at least one of non-small cell lung cancer, small cell lung cancer, or large cell lung cancer; and / or, the breast cancer includes at least one of Her2-positive breast cancer or Her2-negative breast cancer; and / or, the brain tumor includes at least one of glioma, glioblastoma, or astrocytoma; and / or, the neuroendocrine tumor includes at least one of pulmonary neuroendocrine tumor, gastric neuroendocrine tumor, enteroendocrine tumor, or pancreatic neuroendocrine tumor.

[0181] In some embodiments, the activity of the intervention ATR includes at least one of the following:

[0182] Inhibit the interaction between ATR and ATRIP;

[0183] This causes the complex of ATR and ATRIP to dissociate;

[0184] Inhibit the interaction between ATR and TopBP1;

[0185] This causes the complex of ATR and TopBP1 to dissociate;

[0186] Inhibit the interaction between ATR and ETAA1;

[0187] This causes the complex of ATR and ETAA1 to dissociate;

[0188] Degrade at least one of ATR, ATRIP, TopBP1, ETAA1, ATR and ATRIP complex, ATR and TopBP1 complex, or ATR and ETAA1 complex.

[0189] Inhibit the activity of ATR protein;

[0190] Inhibit ATR autophosphorylation;

[0191] Inhibit ATR phosphorylation of at least one of CHK1, p53, SMARCAL1, WRN, or FANCI;

[0192] Repressing the transcription and / or translation of atr genes; and / or

[0193] This can cause deletion and / or mutation of the atr gene.

[0194] In some embodiments, the intervention on mTOR activity includes at least one of the following:

[0195] Inhibit mTOR activity;

[0196] Inhibit the activity of at least one of mTORC1 or mTORC2;

[0197] Inhibit the generation of at least one of mTORC1 or mTORC2;

[0198] To cause at least one of mTORC1 or mTORC2 to dissociate;

[0199] Degrade at least one of mTOR, mTORC1, or mTORC2;

[0200] Repressing the transcription and / or translation of the mtor gene; and / or

[0201] This can cause deletion and / or mutation of the mtor gene.

[0202] In some embodiments, the drug or drug composition includes at least one of protein, polypeptide, nucleic acid, polynucleotide, polynucleotide, antibody or derivative thereof, organic synthetic compound, inorganic compound and / or natural product.

[0203] In some embodiments, the drug or drug composition includes compounds that inhibit ATR activity and mTOR activity.

[0204] In some embodiments, the drug or drug composition includes an ATR inhibitor and an mTOR inhibitor.

[0205] In some implementations, the ATR inhibitor and the mTOR inhibitor may be administered simultaneously, or the ATR inhibitor may be administered first, followed by the mTOR inhibitor at a reasonable interval, such as 5 minutes, 10 minutes, 60 minutes, or 24 hours, or vice versa.

[0206] In some embodiments, the ATR inhibitor comprises at least one of AZD6738, M-4344, M6620, BAY-1895344, RP-3500, M1774, ART0380, SKLB-197, AD1058, or a tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt, or a solvate of a pharmaceutically acceptable salt thereof; and / or

[0207] The mTOR inhibitors include sirolimus, zotalimus, umirolimus, tesimolimus, gidalixetine, Paxalisib, SR-0379, Dactolisib, Oleanin, Olcorolimus, Everolimus, RM-006, desfolimus, SF-1126, KB02-SLF, LY-3023414, TAK-228, AZD2014, CC223, CC-115, RTB-101, Voxtalisib, ME-344, SCC-31, CC-214-2, DHW-208, FD-274, GNE-477, GNE-555, Panulisib, PF-04691503, Piplartine, PP30, PQR514, Torin2, Torkinib, X-387, PQR- 530, Torin1, AZD8055, KU-0063794, BGT-226, Bimiralisib, GDC-0349, Omilise, OSI-027, PF-0469 1502, PKI-179, XL-765, Apitolisib, DS-7423, GNE-493, MKC-1, P-2281, Palomids, PI-103, PWT- 33597, SN-32976, WYE-132, XL-388, PP242, PP30, XL388, WYE-354, WAY-600, WYE-687, AP23573, MLN0128 or at least one of the following: tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt or solvate of a pharmaceutically acceptable salt thereof.

[0208] In some embodiments, the ATR inhibitor is selected from at least one of AZD6738, M-4344, or their tautomers, stereoisomers, enantiomers, diastereomers, racemic mixtures, solvates, isotope derivatives, pharmaceutically acceptable salts, or solvates of pharmaceutically acceptable salts thereof; and / or

[0209] The mTOR inhibitors include at least one of AZD2014, CC223 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

[0210] In some embodiments, the dosage form of the drug or drug composition includes at least one of the following: solution, powder, granule, tablet, sugar-coated, capsule, granule, suspension, syrup, drops, and sublingual tablet.

[0211] In some embodiments, the pharmaceutical composition further includes pharmaceutically acceptable excipients, optionally including at least one of solubilizers, disintegrants, wetting agents, stabilizers, thickeners, diluents, buffers, and flavoring agents.

[0212] use

[0213] Thirdly, this application also provides the use of a drug or pharmaceutical composition capable of intervening in the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR) in the preparation of a drug or pharmaceutical composition for the prevention, relief or treatment of cancer, wherein the drug or pharmaceutical composition inactivates or reduces the activity of ATR and mTOR.

[0214] In some embodiments, the cancer includes at least one of the following: esophageal cancer, gastroesophageal junction cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, melanoma, lung cancer, thyroid cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, brain cancer (including glioma, glioblastoma, astrocytoma, etc.), neuroendocrine tumor, kidney cancer, urothelial carcinoma, bladder cancer, prostate cancer, liver cancer, leukemia, lymphoma, multiple myeloma, sarcoma, or glioma.

[0215] In some embodiments, the esophageal cancer includes at least one of esophageal squamous cell carcinoma or esophageal adenocarcinoma; and / or, the melanoma includes at least one of acral melanoma, mucosal melanoma, or cutaneous melanoma; and / or, the lung cancer includes at least one of non-small cell lung cancer, small cell lung cancer, or large cell lung cancer; and / or, the breast cancer includes at least one of Her2-positive breast cancer or Her2-negative breast cancer; and / or, the brain tumor includes at least one of glioma, glioblastoma, or astrocytoma; and / or, the neuroendocrine tumor includes at least one of pulmonary neuroendocrine tumor, gastric neuroendocrine tumor, enteroendocrine tumor, or pancreatic neuroendocrine tumor.

[0216] In some embodiments, the activity of the intervention ATR includes at least one of the following:

[0217] Inhibit the interaction between ATR and ATRIP;

[0218] This causes the complex of ATR and ATRIP to dissociate;

[0219] Inhibit the interaction between ATR and TopBP1;

[0220] This causes the complex of ATR and TopBP1 to dissociate;

[0221] Inhibit the interaction between ATR and ETAA1;

[0222] This causes the complex of ATR and ETAA1 to dissociate;

[0223] Degrade at least one of ATR, ATRIP, TopBP1, ETAA1, ATR and ATRIP complex, ATR and TopBP1 complex, or ATR and ETAA1 complex.

[0224] Inhibit the activity of ATR protein;

[0225] Inhibit ATR autophosphorylation;

[0226] Inhibit ATR phosphorylation of at least one of CHK1, p53, SMARCAL1, WRN, or FANCI;

[0227] Repressing the transcription and / or translation of atr genes; and / or

[0228] This can cause deletion and / or mutation of the atr gene.

[0229] In some embodiments, the intervention on mTOR activity includes at least one of the following:

[0230] Inhibit mTOR activity;

[0231] Inhibit the activity of at least one of mTORC1 or mTORC2;

[0232] Inhibit the generation of at least one of mTORC1 or mTORC2;

[0233] To cause at least one of mTORC1 or mTORC2 to dissociate;

[0234] Degrade at least one of mTOR, mTORC1, or mTORC2;

[0235] Repressing the transcription and / or translation of the mtor gene; and / or

[0236] This can cause deletion and / or mutation of the mtor gene.

[0237] In some embodiments, the drug or drug composition includes at least one of protein, polypeptide, nucleic acid, polynucleotide, polynucleotide, antibody or derivative thereof, organic synthetic compound, inorganic compound and / or natural product.

[0238] In some embodiments, the drug or drug composition includes compounds that inhibit ATR activity and mTOR activity.

[0239] In some embodiments, the drug or drug composition includes an ATR inhibitor and an mTOR inhibitor.

[0240] In some embodiments, the ATR inhibitor comprises at least one of AZD6738, M-4344, M6620, BAY-1895344, RP-3500, M1774, ART0380, SKLB-197, AD1058, or a tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt, or a solvate of a pharmaceutically acceptable salt thereof; and / or

[0241] The mTOR inhibitors include sirolimus, zotamolimus, umirolimus, tesimolimus, gidalixetine, Paxalisib, SR-0379, Dactolisib, Oleanin Olcorolimus, Everolimus, RM-006, desfolimus, SF-1126, KB02-SLF, LY-3023414, TAK-228, AZD2014, CC223, CC-115, RTB-101, Voxtalisib, ME-344, SCC-31, CC-214-2, DHW-208, FD-274, GNE-477, GNE-555, Panulisib, PF-04691503, Piplartine, PP30, PQR514, Torin2, Torkinib, X-387, and PQR-5. 30. Torin1, AZD8055, KU-0063794, BGT-226, Bimiralisib, GDC-0349, Omilise, OSI-027, PF-04691 502, PKI-179, XL-765, Apitolisib, DS-7423, GNE-493, MKC-1, P-2281, Palomids, PI-103, PWT-3 3597, SN-32976, WYE-132, XL-388, PP242, PP30, XL388, WYE-354, WAY-600, WYE-687, AP23573, MLN0128 or at least one of the following: tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt or solvate of a pharmaceutically acceptable salt thereof.

[0242] In some embodiments, the ATR inhibitor is selected from at least one of AZD6738, M-4344, or their tautomers, stereoisomers, enantiomers, diastereomers, racemic mixtures, solvates, isotope derivatives, pharmaceutically acceptable salts, or solvates of pharmaceutically acceptable salts thereof; and / or

[0243] The mTOR inhibitors include at least one of AZD2014, CC223 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

[0244] In some embodiments, the dosage form of the drug or drug composition includes at least one of the following: solution, powder, granule, tablet, sugar-coated, capsule, granule, suspension, syrup, drops, and sublingual tablet.

[0245] In some embodiments, the pharmaceutical composition further includes pharmaceutically acceptable excipients, optionally including at least one of solubilizers, disintegrants, wetting agents, stabilizers, thickeners, diluents, buffers, and flavoring agents.

[0246] The various embodiments and preferences disclosed above can be combined with each other (as long as they are not inherently contradictory), and all embodiments formed by such combinations are considered as part of the disclosure of this application.

[0247] The exemplary embodiments of this application will now be described with reference to the accompanying drawings, including various details of the embodiments to aid understanding. It should be understood that these are merely exemplary and are in no way intended to limit the scope of protection of this application. The scope of protection of this application is defined only by the claims. Therefore, those skilled in the art will recognize that various changes and modifications can be made to the embodiments described herein without departing from the scope of this application. Similarly, for clarity and brevity, descriptions of well-known functions and structures are omitted in the following description.

[0248] Example

[0249] Unless otherwise specified in this embodiment, the techniques or conditions described in the literature in this field or in accordance with the product instructions shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0250] Example 1: Compound mTOR Inhibition Activity Test

[0251] Using an in-cell Western blotting (ICW) method, the inhibitory effects of AZD6738 (Bidetech Pharmaceuticals, EKU575), M-4344 (Bidetech Pharmaceuticals, CRU153; ETX005), AZD2014 (Bidetech Pharmaceuticals, AMU821), and CC223 (Bidetech Pharmaceuticals, BDC519) on mTOR activity in human esophageal squamous cell carcinoma cells KYSE410 were tested by detecting the phosphorylation levels of substrate S6 (S6(S235 / 236)) of the mTORC1 complex or substrate AKT (S473) of the mTORC2 complex.

[0252] KYSE410 cells (human esophageal cancer cells, Zhejiang Meisen Cell Technology Co., Ltd.) (CTCC-003-0062) cultured in RPMI1640 medium (Gibco, #11875093) (containing 10% FBS and 1% P / S) were cultured at a density of 3 × 10⁶ cells per well. 3 Cells were seeded at a density of 1000 cells / well in 96-well plates. The plates were incubated at 37°C with 5% CO2 for approximately 40 hours. On the third day of the experiment, different concentrations of the test compound (DMSO final concentration 0.1%) were added to each well. After further incubation at 37°C with 5% CO2 for 0.5–2 hours, the following steps were performed: cell fixation with 3.7% formaldehyde, cell permeabilization with Triton X-100, cell blocking with blocking solution, primary antibody incubation, secondary antibody development, plate reading using an Odyssey SA instrument, and statistical analysis of the data using GraphPad Prism software to calculate the IC50. 50 Numerical value.

[0253] The specific experimental steps are as follows:

[0254] 1.1 Compound preparation and cell treatment

[0255] The compound was prepared into a 10 mM stock solution using DMSO; in a 96-well plate, the compound was serially diluted with DMSO at 5-fold concentrations to obtain compounds with concentrations of 10 mM, 2 mM, 0.4 mM, 80 μM, 16 μM, 3.2 μM, 0.64 μM, 128 nM, and 25.6 nM.

[0256] Preparation of working solution: Add 98 μL of culture medium to a 96-well plate, then add 2 μL of serially diluted compound or DMSO, seal the plate, vortex to mix, centrifuge for 20 seconds to drop the solution to the bottom of the tube, and set aside for use.

[0257] Take KYSE410 cells, add 5.3 μL of working solution to each well, and repeat the compound / DMSO solution twice. Return the cells to the incubator for incubation.

[0258] 1.2 Cell fixation and antibody incubation

[0259] Aspirate the culture medium from the 96-well plate; rinse each well once with 150 μL of 1X PBS; add 150 μL of fixative (37% formaldehyde diluted in 1X PBS to obtain 3.7% formaldehyde), fix at room temperature for 20 min, and aspirate; add 100 μL of permeabilization buffer (1X PBS, 0.1% Triton™ X-100 (SIGMA-ALDRICH, T8787)), incubate at room temperature for 10 min, and aspirate; rinse with 150 μL of 1X PBS, incubate on a microplate shaker at room temperature for 5 min, repeat 4 times; add 100 μL of... (TBS) Blocking Buffer (LI-COR, 926-68070), incubate on a shaker at room temperature for 1.5 hours; after drying, add 50 μL of primary antibody (set up two control wells: DMSO treatment without primary antibody, and 50 μL of blocking buffer), and incubate overnight at 4°C. The primary antibody formulation is shown in the table below:

[0260] Remove the microplate from the 4°C freezer and allow it to return to room temperature. After drying, rinse with 150 μL of 1X TBS / T (diluted with ddH2O to 10X TBS / T and supplemented with 0.05% Tween-20). Incubate on a microplate shaker at room temperature for 5 min. Repeat this step 4 times. Add 50 μL of secondary antibody and incubate on a shaker at room temperature in the dark for 1 hour. The formulation of the secondary antibody is shown in the table below.

[0261] Spike dry, rinse with 150 μL 1X TBST, incubate on a microplate shaker at room temperature for 5 min, repeat 4 times, spin dry; read the plate with LI-COR Odyssey SA and export the data.

[0262] 1.3 Statistical Analysis

[0263] For p-S6 and p-AKT, the secondary antibody background mean was first calculated (for the control wells with DMSO but no primary antibody). Corrected intensity = original data minus secondary antibody background mean. Normalization of corrected intensity = 800nm ​​value / 700nm value in the same well. Inhibition rate = 100% - Signal(cmpd) / Signal(DMSO) * 100%. Finally, the compound concentration and inhibition rate data were imported into GraphPad Prism for non-linear regression analysis and curve fitting to obtain the IC50. 50Value(Y=Bottom+(Top-Bottom) / (1+((X^HillSlope) / (IC 50 ^HillSlope)))).

[0264] Experimental results

[0265] Table 1. mTOR inhibitory activity of the compounds Note: " / " indicates that the experimental data could not be curve-fitted to calculate IC. 50 The numerical value can be understood as the compound having no inhibitory activity against the target.

[0266] Table 1 shows that in the human esophageal cancer cell line KYSE410 experimental system, AZD2014 and CC223 exhibited inhibitory activity against both mTORC1 and mTORC2; AZD6738 showed no inhibitory activity against either mTORC1 or mTORC2; and M-4344 showed an IC50 of [missing information - likely referring to an IC50 value] against mTORC1. 50 The inhibitory activity is close to the μM level, indicating weak inhibitory capacity, especially for mTORC2. Therefore, AZD2014 and CC223 can be considered inhibitors of mTORC1 / 2, while AZD6738 and M-4344 have no mTORC1 / 2 inhibitory activity.

[0267] Example 2: Compound ATR Inhibition Activity Test

[0268] Using an in-cell Western blotting (ICW) method, the inhibitory effects of AZD6738, M-4344, AZD2014, and CC223 on ATR kinase activity in human colon cancer cells HT29 were tested by detecting the phosphorylation level of the ATR substrate CHK1 (Ser345).

[0269] The specific steps of Example 2 are similar to those of Example 1, except that:

[0270] HT29 cells were cultured in McCoy's 5A medium (Meilunbio, PWL006) at a concentration of 1 × 10⁶ cells per well. 4 Cells were seeded at a density of 100 mM in 96-well plates. 4-NQO (4-nitroquinoline-N-oxide) was dissolved in DMSO and thoroughly mixed to obtain a 6 mM concentration. This was then diluted 100-fold with complete culture medium to obtain a 60 μM 4-NQO working solution. On the third day of the experiment, different concentrations of the test compound (DMSO final concentration 0.1%, same as in Example 1) were added to each well. After incubation at 37°C and 5% CO2 for 1 h, DNA damage was stimulated in the cells by adding a 3 μM 4-NQO working solution. Subsequent steps were then performed as described in Example 1.

[0271] The formulation of the primary antibody is shown in the table below:

[0272] In the data statistical analysis section, the mean value of wells without 4-NQO treatment (DMSO without 4-NQO) was first calculated. Corrected intensity = original data minus the mean value of wells without 4-NQO treatment. Normalization of corrected intensity = 800nm ​​value / 700nm value of the same well. Inhibition rate = 100% - Signal(cmpd) / Signal(DMSO) * 100%. Finally, the compound concentration and inhibition rate data were imported into GraphPad Prism, and non-linear regression analysis and curve fitting were performed to obtain IC50. 50 Value(Y=Bottom+(Top-Bottom) / (1+((X^HillSlope) / (IC 50 ^HillSlope)))).

[0273] Experimental results

[0274] Table 2. ATR inhibitory activity of the compounds Note: " / " indicates that the experimental data could not be curve-fitted to calculate IC. 50 A value or IC50 > 100000 nM can be interpreted as the compound having no inhibitory activity against the target.

[0275] As shown in Table 2, in the experimental system for inducing DNA damage in human colon cancer cells HT29 by 4-NQO, AZD6738 and M-4344 both exhibited strong ATR inhibitory activity; while AZD2014 and CC223 showed no ATR inhibitory activity. Therefore, AZD6738 and M-4344 can be considered ATR inhibitors, while AZD2014 and CC223 are not.

[0276] As can be seen from Examples 1 and 2, for the dual-target strategy of ATR and mTOR involved in this application, AZD2014 and CC223 are single-target inhibitors of mTORC1 / 2, while AZD6738 and M-4344 are single-target inhibitors of ATR.

[0277] Example 3: Antitumor efficacy test of the combination of M-4344 and AZD2014 in a KYSE410 mouse subcutaneous xenograft model

[0278] 3.1 Laboratory animals:

[0279] NOD SCID mice (Shanghai Lingchang Biotechnology Co., Ltd., female, 6 weeks old) were housed in the experimental environment for 3-7 days after arrival before the experiment began. The animals were housed in an SPF-grade cleanroom environment.

[0280] 3.2 Modeling and Grouping

[0281] KYSE410 cells were cultured in RPMI-1640 medium (BasalMedia, L210KJ) containing 10% FBS. KYSE410 cells in the exponential growth phase were collected, the required total cell count was calculated, and then the cells were resuspended in an appropriate volume of PBS and matrix gel (1:1) to a cell suspension concentration of 1×10⁻⁶ cells / cells. 7 0.2 mL of cells per mouse was administered via subcutaneous injection on the right side of NOD SCID mice, with 0.2 mL injected per mouse.

[0282] On the day of vaccination (Day 0), 0.2 mL (1×10⁻⁶) was subcutaneously injected into the right back of each animal. 7 A subcutaneous xenograft tumor model was established using a suspension of KYSE410 cells. The health status and tumor growth of the mice were observed daily after cell inoculation. On day 5 post-inoculation, the tumors had grown to an average volume of 159.2 mm. 3 The animals were randomly divided into groups of 8 according to tumor volume and started treatment. The dosing frequency is shown in the table below: Note: "N" represents the number of animals; "QD" means once a day; Group G4 is the combined drug group, and the drug administration order is to administer M-4344 first, followed by AZD2014 0.5h to 1h later.

[0283] The preparation process of the compounds in each group is shown below:

[0284] M-4344: Weigh 4.05 mg of M-4344 powder, add 3 mL of 15% Captisol solution, adjust the pH to 3.5 ± 0.10 with 0.1 M HCl, vortex and sonicate to mix, and finally add the remaining 15% Captisol to obtain 4 mL of 1 mg / mL solution.

[0285] AZD2014: Weigh 4.03 mg of AZD2014 powder, add 4 mL of 30% Captisol solution, and vortex sonicate to mix well to obtain 4 mL of 1 mg / mL solution.

[0286] 3.3 Experimental Observation

[0287] Monitor the animals' health and mortality daily, and measure and calculate tumor volume twice a week. Tumor volume calculation formula: Tumor volume (mm²) 3= 0.5 × (tumor long diameter × tumor short diameter) 2 The formula for calculating relative tumor volume is: Relative tumor volume = Average tumor volume after treatment / Average tumor volume at the time of grouping. Throughout the study, the entire drug administration process, as well as tumor and body weight measurements, were performed in a clean bench.

[0288] 3.4 Data Processing

[0289] Tumor volume inhibition rate (TGI) TV (%): TGI% = (1 - ΔT / ΔC) × 100%; where ΔC is the tumor volume C of the control group. t -C0, where C0 is the average tumor volume of the control group at the time of grouping, C t The mean tumor volume in the control group after treatment is ΔT; ΔT represents the tumor volume in the treatment group (T). t -T0, where T0 is the average tumor volume of the treatment group at the time of grouping, T t The mean tumor volume in the treatment group after treatment.

[0290] Relative tumor inhibition rate (TGI) RTV (%): TGI RTV =1-T RTV / C RTV (%). T RTV / C RTV % represents the relative tumor proliferation rate, which is the percentage of tumor volume in the treatment group and the control group at a certain time point. T RTV and C RTV The relative tumor volume (TV) of the treatment group and the control group at a specific time point are respectively.

[0291] The calculation formula is as follows: T RTV : Mean RTV in the treatment group; C RTV : Average RTV of the control group; RTV = V t / V0, where V0 is the tumor volume of the animal at the time of grouping, V t The tumor volume of the animal after treatment.

[0292] Tumor volume and animal weight results are expressed as Mean ± SEM (mean standard error). Statistical comparative analysis of tumor volume was performed between different groups. All statistical analyses were performed online using Meritudio; the statistical methods used are described at https: / / doi.org / 10.1186 / s12885-019-5907-7 and https: / / doi.org / 10.1158 / 2767-9764.CRC-23-0243, respectively. The Wilcox.test nominal or t.test nominal methods were used to compare whether there were significant differences in tumor volume between groups, where p ≥ 0.05 was considered not significant, p < 0.05 was considered significant, and p < 0.01 was considered highly significant. GraphPad Prism software was used to plot the tumor volume and weight change curves.

[0293] Experimental results

[0294] Table 3. Analysis and comparison of mean tumor volume among groups on day 21 after grouping. Note: P value a The tumor volume of groups G2, G3, and G4 was compared with that of group G1; tumor volume of G2 vs G4: p = 0.0285; tumor volume of G3 vs G4: p = 0.0016.

[0295] As shown in Figure 1 and Table 3, on day 21 after grouping, the mean tumor volume of groups G2, G3, and G4 was significantly smaller than that of the control group G1. Specifically, compared with the G2 (M-4344, an ATR inhibitor) monotherapy group or the G3 (AZD2014, an mTOR inhibitor) monotherapy group, the combination therapy group G4 (M-4344+ZAD2014) showed significantly lower mean tumor volume and relative tumor volume, and a higher tumor volume inhibition rate (TGI). TV ) and relative tumor inhibition rate (TGI) RTV Both were significantly improved, with statistically significant differences (p<0.05). This result indicates that the dual-target drug combination of M-4344 and AZD2014 demonstrates a significant efficacy advantage in the human esophageal cancer KYSE410 subcutaneous xenograft model compared to a single-target strategy.

[0296] As shown in Figure 2, compared with the control group, there were no significant changes in body weight in mice in groups G2, G3, and G4 after administration of the compounds and their combinations described in this application; among them, mice in group G4 tolerated the treatment well, and their body weight increased steadily. At the end of the experiment, gross necropsy was performed on mice in all groups, and no abnormalities were found. This indicates that the dual-target drug combination of M-4344 and AZD2014 has good safety in the human esophageal cancer KYSE410 subcutaneous xenograft model.

[0297] Example 4: Antitumor efficacy test of the combination of M-4344 and AZD2014 in a mouse subcutaneous xenograft model of human large cell lung cancer NCI-H460.

[0298] 4.1 Laboratory animals:

[0299] BALB / c Nude mice (female, 5-6 weeks old, Shanghai Jihui Experimental Animal Breeding Co., Ltd.) were housed in the experimental environment for 3 days after arrival before the start of the experiment. The animals were housed in an SPF-grade cleanroom environment.

[0300] 4.2 Modeling and Grouping

[0301] NCI-H460 cells (catalog number: NA20230627) were cultured in RPMI-1640 medium (Gibco, catalog number: C11875500BT) containing 10% FBS. NCI-H460 cells in the exponential growth phase were collected, the required total cell count was calculated, and then the cells were resuspended in appropriate volumes of PBS and Matrigel (PBS to Matrigel volume ratio 1:1) to a cell suspension concentration of 1.69 × 10⁻⁶ cells / year. 6 0.1 mL of cells per mouse was administered via subcutaneous injection into the right back of NOD SCID mice, with each mouse receiving 0.1 mL.

[0302] On the day of vaccination (Day 0), 0.1 mL (1.69 × 10⁻⁶) was subcutaneously injected into the right back of each animal. 6 A subcutaneous xenograft tumor model was established using a suspension of NCI-H460 cells. The health status and tumor growth of the mice were observed daily after cell inoculation. The model was established when the tumor reached an average volume of 150 mm². 3 The mice were randomly divided into groups of eight based on tumor volume and body weight, and treatment was initiated at the appropriate times. The dosing frequencies are shown in Table 4 below. Note: "N" represents the number of animals; "QD" means once a day; Group G8 is the combined drug group, and the drug administration order is to administer AZD6738 first, followed by CC223 0.5h to 1h later.

[0303] The methods for preparing the compound solutions in each group are as follows:

[0304] 4.3 Experimental Observation

[0305] After tumor cell inoculation, routine monitoring included tumor growth and the effect of drug administration on the animal's normal behavior, with tumor measurements and volume calculations performed twice a week.

[0306] Formula for calculating tumor volume: Tumor volume (mm) 3 = 0.5 × (tumor long diameter × tumor short diameter) 2 ).

[0307] The formula for calculating relative tumor volume is: Relative tumor volume = Average tumor volume after treatment / Average tumor volume at the time of grouping.

[0308] Weight change rate (%) = (Weight on the day of administration - Weight at the time of first administration) / Weight at the time of first administration * 100%.

[0309] Throughout the study, the entire drug administration process, as well as tumor and weight measurements, were performed in a clean bench.

[0310] 4.4 Data Processing

[0311] Use Excel to calculate tumor volume and weight change rate.

[0312] Formula for calculating tumor volume: Tumor volume (mm) 3 )=1 / 2×(D×d 2 (where D represents the major axis and d represents the minor axis).

[0313] Weight change rate (%) = (Weight on the day of administration - Weight at the time of first administration) / Weight at the time of first administration * 100%.

[0314] The relative tumor inhibition rate, TGI (%), is calculated as follows: TGI% = (1-T / C) × 100%. (T and C are the relative tumor volumes (RTV) of the treatment group and the control group at a specific time point, respectively.)

[0315] Relative tumor proliferation rate (T / C%): T / C% = TRTV / CRTV × 100% (TRTV: mean RTV in the treatment group; CRTV: mean RTV in the solvent control group; RTV = Vt / V0, where V0 is the tumor volume of the animal at the time of grouping, and Vt / V0 is the tumor volume of the animal after treatment).

[0316] The data were analyzed using GraphPad Prism 8.0 software, and a two-way ANOVA was used to determine whether the differences in the data were significant.

[0317] 4.5 Experimental Results

[0318] The analysis of the antitumor effects of the test drugs is shown in Table 5, and the tumor growth of mice in each group is shown in Figure 3.

[0319] Table 5. Efficacy analysis of each group in the NCI-H460 human large cell lung cancer model. Note: 1. G2 vs G4: p = 0.0168 # G3 vs G4: p = 0.0085 ## 2. ns: P>0.05, no significant difference; 3. "* / # "**" indicates a significant difference; 4. "** / ## "Indicates a highly significant difference."

[0320] As shown in Figure 3 and Table 5, on day 20 after grouping, the mean tumor volume of groups G6, G7, and G8 was significantly smaller than that of the control group G5. Specifically, compared with the G6 (AZD6738, an ATR inhibitor) monotherapy group or the G7 (CC223, an mTOR inhibitor) monotherapy group, the combination therapy group G8 (AZD67384+CC223) showed significantly lower mean tumor volume and relative tumor volume, and a higher tumor volume inhibition rate (TGI). TV ) and relative tumor inhibition rate (TGI) RTV All parameters showed significant improvement, with statistically significant differences (p<0.05). This result indicates that the dual-target drug combination of AZD6738 and CC223 demonstrates a significant efficacy advantage in the NCI-H460 human large cell lung cancer subcutaneous xenograft model compared to a single-target strategy.

[0321] As shown in Figure 4, compared with the control group, there were no significant changes in body weight in mice in groups G6, G7, and G8 after administration of the compounds and combinations thereof; no mice died. This indicates that the dual-target drug combination of M-4344 and AZD2014 has good safety in the human esophageal cancer KYSE410 subcutaneous xenograft model.

[0322] Example 5: Anti-proliferation effect of ATR inhibitors (AZD6738, M4344) in combination with two mTOR inhibitors (AZD2014, CC223) in mouse skin melanoma cells B16-F10.

[0323] Cell proliferation was assessed using the CellTiter-Glo luminescent viable cell assay. This kit utilizes luciferase for detection; the luminescence process of luciferase requires ATP, and the amount of ATP is directly proportional to the number of viable cells. Therefore, the luminescence value measured after adding CellTiter-Glo reagent to the cell culture system reflects the number of viable cells. After calculating the cell proliferation inhibition rate using EXCEL, the data were analyzed using Synergyfinder software to obtain the dose-response matrix of the compound combinations. The synergistic effect score of the compound combinations was calculated using CompSyn within a certain concentration range.

[0324] 5.1 Test Materials

[0325] Test compound:

[0326] Preparation of compound stock solution: The test compound powder was stored in a refrigerator at 4°C. Before testing, a 10 mM stock solution was prepared with 100% DMSO and stored in a refrigerator at -80°C.

[0327] Cell lines:

[0328] Main reagents:

[0329] 5.2 Test Methods

[0330] B16-F10 cells (mouse skin melanoma cell line) or NCI-H460 cells (human large cell lung cancer cell line) in the logarithmic growth phase were harvested and counted using a platelet counting chamber; cell viability was simultaneously assessed using the trypan blue rejection assay to ensure a viability of over 90%. The cell suspension concentration was adjusted to 2 × 10⁻⁶ cells / mL. 4 / mL. Add 100 μL of cell suspension to each well of a 96-well plate to obtain 2000 cells per well. Incubate the cells in the 96-well plate overnight at 37°C, 5% CO2, and 95% humidity.

[0331] Remove the diluted stock solution from the -20°C freezer, thaw at room temperature, and then vortex to mix thoroughly. Dilute the stock solution to the highest concentration required for each cell type, and then perform serial dilutions of the compound in a 96-well qPCR plate. Add 98 μL of complete culture medium to each well of the 96-well qPCR plate, followed by 2 μL of the serially diluted test compound or DMSO. Seal the qPCR plate with sealing film, vortex to mix, and centrifuge for 30 seconds to remove the solution to the bottom of the qPCR plate. Remove the cells that have been cultured overnight, and add 5.6 μL of the compound or DMSO working solution to each well. Gently tap the side of the plate to mix. Return the 96-well cell culture plate to an incubator at 37°C, 5% CO2, and 95% relative humidity for 3 days.

[0332] Melting CellTiter-Glo reagent ( (Luminescent Cell Viability Assay, Promega, G7573) and equilibrate the cell plate to room temperature for 30 minutes. Add an equal volume of CellTiter-Glo solution to each well and shake on a track-mounted shaker for 2 minutes to lyse the cells. Place the cell plate at room temperature for 10 minutes to stabilize the cold light signal, and detect the chemiluminescence signal (Luminescence, RLU) on a microplate reader (ESCO, CCL-170T-8).

[0333] Data analysis used the following formula to calculate the inhibition rate (IR) of the detected compound: IR (%) = (1 - [compound OD450 - blank control OD450] / (solvent control OD450 - blank control OD450)] * 100%. The dose-response matrix of the two compounds was obtained using Synergyfinder analysis. Finally, based on the dose-response matrix, the CI value of the two drugs in combination was calculated using CompuSyn software within a certain concentration range.

[0334] 5.3 Experimental Results

[0335] Figure 5 shows the inhibitory effect of the combination of compounds M4344 and AZD2014 in NCI-H460 cells. The values ​​in the dose-response matrix are as follows:

[0336] Figure 6 shows the inhibitory effect of the combination of compounds AZD6738 and AZD2014 in NCI-H460 cells. The values ​​in the dose-response matrix are as follows:

[0337] Figure 7 shows the inhibitory effect of the combination of compounds AZD6738 and CC223 in NCI-H460 cells. The values ​​in the dose-response matrix are as follows:

[0338] Figure 8 shows the inhibitory effect of the combination of compounds M4344 and CC223 in NCI-H460 cells. The values ​​in the dose-response matrix are as follows:

[0339] Figure 9 shows the inhibitory effect of the combination of compounds M4344 and AZD2014 in B16-F10 cells. The values ​​in the dose-response matrix are shown below:

[0340] Figure 10 shows the inhibitory effect of the combination of compounds CC223 and M4344 in B16-F10 cells. The values ​​in the dose-response matrix are shown below:

[0341] Figure 11 shows the inhibitory effect of the combination of compounds AZD6738 and AZD2014 in B16-F10 cells. The values ​​in the dose-response matrix are as follows:

[0342] Figure 12 shows the inhibitory effect of the combination of compounds AZD6738 and CC223 in B16-F10 cells. The values ​​in the dose-response matrix are as follows:

[0343] Figures 5-8 show that in NCI-H460 cells, the combinations of AZD2014 (62.5–1000 nM) with M4344 (24.69–2000 nM), AZD2014 (125–2000 nM) with AZD6738 (156.25–2500 nM), and CC223 (24.69–2000 nM) with AZD6738 (312.5–5000 nM) all yielded CI values ​​less than 1, indicating a synergistic effect. The combination of CC223 with M4344 only showed a synergistic effect in a smaller concentration range of 24.69–74.07 nM.

[0344] Figures 9-12 show that in B16-F10 cells, the combinations of AZD2014 (31.25–500 nM) with M4344 (24.69–2000 nM), CC223 (625–10000 nM) with M4344 (24.69–666.66 nM), AZD2014 (62.5–1000 nM) with AZD6738 (24.69–2000 nM), and CC223 (312.25–5000 nM) with AZD6738 (24.69–2000 nM) all yielded CI values ​​less than 1, indicating a synergistic effect.

[0345] In this study, the combination of two ATR inhibitors (AZD6738, M4344) and two mTOR inhibitors (AZD2014, CC223) in B16-F10 and NCI-H460 cells showed a trend of synergistic anti-cell proliferation effects.

[0346] In summary, compared to using ATR inhibitors or mTOR inhibitors alone, simultaneously inhibiting the activity of both ATR and mTOR targets has a synergistic effect, resulting in a stronger tumor-suppressing effect, while also exhibiting better safety. This means that the dual-target strategy of ATR and mTOR has a larger therapeutic window in cancer treatment than a single-target strategy; this has significant advantages for clinical translation and application development.

Claims

1. A medicament or pharmaceutical composition for the prevention, relief, or treatment of cancer, characterized in that, The drug or drug composition can interfere with the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR), thereby inactivating or reducing the activity of ATR and mTOR.

2. The medicament or pharmaceutical composition according to claim 1, wherein, The cancers mentioned include at least one of the following: esophageal cancer, esophagogastric junction cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, melanoma, lung cancer, thyroid cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, brain cancer, neuroendocrine tumor, kidney cancer, urothelial carcinoma, bladder cancer, prostate cancer, liver cancer, leukemia, lymphoma, multiple myeloma, sarcoma, or glioma.

3. The medicament or pharmaceutical composition according to claim 2, wherein, The esophageal cancer includes at least one of esophageal squamous cell carcinoma or esophageal adenocarcinoma; and / or, the melanoma includes at least one of acral melanoma, mucosal melanoma, or cutaneous melanoma; and / or, the lung cancer includes at least one of non-small cell lung cancer, small cell lung cancer, or large cell lung cancer; and / or, the breast cancer includes at least one of Her2-positive breast cancer or Her2-negative breast cancer; and / or, the brain tumor includes at least one of glioma, glioblastoma, or astrocytoma; and / or, the neuroendocrine tumor includes at least one of pulmonary neuroendocrine tumor, gastric neuroendocrine tumor, enteroendocrine tumor, or pancreatic neuroendocrine tumor.

4. The medicament or pharmaceutical composition according to any one of claims 1-3, wherein, The activity of the intervention ATR includes at least one of the following groups: Inhibit the interaction between ATR and ATRIP; This causes the complex of ATR and ATRIP to dissociate; Inhibit the interaction between ATR and TopBP1; This causes the complex of ATR and TopBP1 to dissociate; Inhibit the interaction between ATR and ETAA1; This causes the complex of ATR and ETAA1 to dissociate; Degrade at least one of ATR, ATRIP, TopBP1, ETAA1, ATR and ATRIP complex, ATR and TopBP1 complex, or ATR and ETAA1 complex. Inhibit the activity of ATR protein; Inhibit ATR autophosphorylation; Inhibit ATR phosphorylation of at least one of CHK1, p53, SMARCAL1, WRN, or FANCI; Repressing the transcription and / or translation of atr genes; and / or This can cause deletion and / or mutation of the atr gene.

5. The medicament or pharmaceutical composition according to any one of claims 1-4, wherein, The intervention on mTOR activity includes at least one of the following groups: Inhibit mTOR activity; Inhibits the activity of at least one of mTORC1 or mTORC2; Inhibit the generation of at least one of mTORC1 or mTORC2; To cause at least one of mTORC1 or mTORC2 to dissociate; Degrade at least one of mTOR, mTORC1, or mTORC2; Repressing the transcription and / or translation of the mtor gene; and / or This can cause deletion and / or mutation of the mtor gene.

6. The medicament or pharmaceutical composition according to any one of claims 1-5, wherein, The drug or drug composition comprises at least one of the following: protein, polypeptide, nucleic acid, polynucleotide, polynucleotide, antibody or derivative thereof, organic synthetic compound, inorganic compound and / or natural product.

7. The medicament or pharmaceutical composition according to any one of claims 1-6, wherein, The drug or drug composition includes compounds that inhibit ATR activity and mTOR activity.

8. The medicament or pharmaceutical composition according to any one of claims 1-7, wherein, The drug or drug composition includes ATR inhibitors and mTOR inhibitors.

9. The medicament or pharmaceutical composition according to claim 8, wherein, The ATR inhibitors include at least one of AZD6738, M6620, M-4344, BAY-1895344, RP-3500, M1774, ART0380, SKLB-197, AD1058, or their tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotopic derivatives, pharmaceutically acceptable salts, or solvates of pharmaceutically acceptable salts thereof; and / or The mTOR inhibitors include sirolimus, zotalimus, umirolimus, tesimolimus, gidalixetine, Paxalisib, SR-0379, Dactolisib, Oleanin, Olcorolimus, Everolimus, RM-006, desfolimus, SF-1126, KB02-SLF, LY-3023414, TAK-228, AZD2014, CC223, CC-115, RTB-101, Voxtalisib, ME-344, SCC-31, CC-214-2, DHW-208, FD-274, GNE-477, GNE-555, Panulisib, PF-04691503, Piplartine, PP30, PQR514, Torin2, Torkinib, X-387, PQR- 530, Torin1, AZD8055, KU-0063794, BGT-226, Bimiralisib, GDC-0349, Omilise, OSI-027, PF-0469 1502, PKI-179, XL-765, Apitolisib, DS-7423, GNE-493, MKC-1, P-2281, Palomids, PI-103, PWT- 33597, SN-32976, WYE-132, XL-388, PP242, PP30, XL388, WYE-354, WAY-600, WYE-687, AP23573, MLN0128 or at least one of the following: tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt or solvate of a pharmaceutically acceptable salt thereof.

10. The medicament or pharmaceutical composition according to claim 8 or 9, wherein, The ATR inhibitor is selected from at least one of AZD6738, M-4344 or its tautomers, stereoisomers, enantiomers, diastereomers, racemic mixtures, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof; and / or The mTOR inhibitors include at least one of AZD2014, CC223 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

11. The medicament or pharmaceutical composition according to any one of claims 1-10, wherein, The dosage form of the drug or drug composition includes at least one of the following: solution, powder, granule, tablet, sugar-coated, capsule, granule, suspension, syrup, drops, and sublingual tablet.

12. The medicament or pharmaceutical composition according to any one of claims 1-11, wherein, The pharmaceutical composition further includes pharmaceutically acceptable excipients, optionally including at least one of solubilizers, disintegrants, wetting agents, stabilizers, thickeners, diluents, buffers, and flavoring agents.

13. A method of preventing, alleviating or treating cancer, characterized in that, The method comprises administering to a subject in need a therapeutically effective dose of a drug or pharmaceutical composition capable of intervening in the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR), said drug or pharmaceutical composition inactivating or reducing the activity of ATR and mTOR.

14. The method of claim 13, wherein, The cancers mentioned include at least one of the following: esophageal cancer, esophagogastric junction cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, melanoma, lung cancer, thyroid cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, brain cancer, neuroendocrine tumor, kidney cancer, urothelial carcinoma, bladder cancer, prostate cancer, liver cancer, leukemia, lymphoma, multiple myeloma, sarcoma, or glioma.

15. The method of claim 14, wherein, The esophageal cancer includes at least one of esophageal squamous cell carcinoma or esophageal adenocarcinoma; and / or, the melanoma includes at least one of acral melanoma, mucosal melanoma, or cutaneous melanoma; and / or, the lung cancer includes at least one of non-small cell lung cancer, small cell lung cancer, or large cell lung cancer; and / or, the breast cancer includes at least one of Her2-positive breast cancer or Her2-negative breast cancer; and / or, the brain tumor includes at least one of glioma, glioblastoma, or astrocytoma; and / or, the neuroendocrine tumor includes at least one of pulmonary neuroendocrine tumor, gastric neuroendocrine tumor, enteroendocrine tumor, or pancreatic neuroendocrine tumor.

16. The method of any one of claims 13-15, wherein, The activity of the intervention ATR includes at least one of the following groups: Inhibit the interaction between ATR and ATRIP; This causes the complex of ATR and ATRIP to dissociate; Inhibit the interaction between ATR and TopBP1; This causes the complex of ATR and TopBP1 to dissociate; Inhibit the interaction between ATR and ETAA1; This causes the complex of ATR and ETAA1 to dissociate; Degrade at least one of ATR, ATRIP, TopBP1, ETAA1, ATR and ATRIP complex, ATR and TopBP1 complex, or ATR and ETAA1 complex. Inhibit the activity of ATR protein; Inhibit ATR autophosphorylation; Inhibit ATR phosphorylation of at least one of CHK1, p53, SMARCAL1, WRN, or FANCI; Repressing the transcription and / or translation of the atr gene; and / or This can cause deletion and / or mutation of the atr gene.

17. The method of any one of claims 13-16, wherein, The intervention on mTOR activity includes at least one of the following groups: Inhibit mTOR activity; Inhibit the activity of at least one of mTORC1 or mTORC2; Inhibit the generation of at least one of mTORC1 or mTORC2; To cause at least one of mTORC1 or mTORC2 to dissociate; Degrade at least one of mTOR, mTORC1, or mTORC2; Repressing the transcription and / or translation of the mtor gene; and / or This can cause deletion and / or mutation of the mtor gene.

18. The method of any one of claims 13-17, wherein, The drug or drug composition comprises at least one of the following: protein, polypeptide, nucleic acid, polynucleotide, polynucleotide, antibody or derivative thereof, organic synthetic compound, inorganic compound and / or natural product.

19. The medicament or pharmaceutical composition according to any one of claims 13-18, wherein, The drug or drug composition includes compounds that inhibit ATR activity and mTOR activity.

20. The method of any one of claims 13-19, wherein, The drug or drug composition includes ATR inhibitors and mTOR inhibitors.

21. The method according to claim 20, wherein, The ATR inhibitors include at least one of AZD6738, M-4344, M6620, BAY-1895344, RP-3500, M1774, ART0380, SKLB-197, AD1058, or their tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotopic derivatives, pharmaceutically acceptable salts, or solvates of pharmaceutically acceptable salts thereof; and / or The mTOR inhibitors include sirolimus, zotamolimus, umirolimus, tesimolimus, gidalixetine, Paxalisib, SR-0379, Dactolisib, EVEROLIMUS, Oleandrin, Olcorolimus, Everolimus, RM-006, desfolimus, SF-1126, KB02-SLF, LY-3023414, TAK-228, AZD2014, CC223, CC-115, RTB-101, Voxtalisib, ME-344, SCC-31, CC-214-2, DHW-208, FD-274, GNE-477, GNE-555, Panulisib, PF-04691503, Piplartine, PP30, PQR514, Torin2, Torkinib, and X-387. , PQR-530, Torin1, AZD8055, KU-0063794, BGT-226, Bimiralisib, GDC-0349, Omilise, OSI-027, PF-0 4691502, PKI-179, XL-765, Apitolisib, DS-7423, GNE-493, MKC-1, P-2281, Palomids, PI-103, PW T-33597, SN-32976, WYE-132, XL-388, PP242, PP30, XL388, WYE-354, WAY-600, WYE-687, AP23573, MLN0128 or at least one of the following: tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt or solvate of a pharmaceutically acceptable salt thereof.

22. The method according to claim 20 or 21, wherein, The ATR inhibitor is selected from at least one of AZD6738, M-4344 or its tautomers, stereoisomers, enantiomers, diastereomers, racemic mixtures, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof; and / or The mTOR inhibitors include at least one of AZD2014, CC223 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

23. The method of any one of claims 13-22, wherein, The dosage form of the drug or drug composition includes at least one of the following: solution, powder, granule, tablet, sugar-coated, capsule, granule, suspension, syrup, drops, and sublingual tablet.

24. The method of any one of claims 13-23, wherein, The pharmaceutical composition further includes pharmaceutically acceptable excipients, optionally including at least one of solubilizers, disintegrants, wetting agents, stabilizers, thickeners, diluents, buffers, and flavoring agents.

25. Use of a drug or pharmaceutical composition capable of interfering with the activity of ataxia telangiectasia mutated and Rad3-related kinase (ATR) and mammalian target of rapamycin (mTOR) in the preparation of a drug or pharmaceutical composition for the prevention, relief or treatment of cancer, wherein the drug or pharmaceutical composition inactivates or reduces the activity of ATR and mTOR.

26. The use of claim 25, wherein, The cancers mentioned include at least one of the following: esophageal cancer, esophagogastric junction cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, bile duct cancer, melanoma, lung cancer, thyroid cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, brain cancer (including glioma, glioblastoma, astrocytoma, etc.), neuroendocrine tumors, kidney cancer, urothelial carcinoma, bladder cancer, prostate cancer, liver cancer, leukemia, lymphoma, multiple myeloma, sarcoma, or glioma.

27. Use according to claim 25 or 26, wherein, The esophageal cancer includes at least one of esophageal squamous cell carcinoma or esophageal adenocarcinoma; and / or, the melanoma includes at least one of acral melanoma, mucosal melanoma, or cutaneous melanoma; and / or, the lung cancer includes at least one of non-small cell lung cancer, small cell lung cancer, or large cell lung cancer; and / or, the breast cancer includes at least one of Her2-positive breast cancer or Her2-negative breast cancer; and / or, the brain tumor includes at least one of glioma, glioblastoma, or astrocytoma; and / or, the neuroendocrine tumor includes at least one of pulmonary neuroendocrine tumor, gastric neuroendocrine tumor, enteroendocrine tumor, or pancreatic neuroendocrine tumor.

28. The use of any one of claims 25-27, wherein, The activity of the intervention ATR includes at least one of the following groups: Inhibit the interaction between ATR and ATRIP; This causes the complex of ATR and ATRIP to dissociate; Inhibit the interaction between ATR and TopBP1; This causes the complex of ATR and TopBP1 to dissociate; Inhibit the interaction between ATR and ETAA1; This causes the complex of ATR and ETAA1 to dissociate; Degrade at least one of ATR, ATRIP, TopBP1, ETAA1, ATR and ATRIP complex, ATR and TopBP1 complex, or ATR and ETAA1 complex. Inhibit the activity of ATR protein; Inhibit ATR autophosphorylation; Inhibit ATR phosphorylation of at least one of CHK1, p53, SMARCAL1, WRN, or FANCI; Repressing the transcription and / or translation of atr genes; and / or This can cause deletion and / or mutation of the atr gene.

29. The use according to any one of claims 25-28, wherein, The intervention on mTOR activity includes at least one of the following groups: Inhibit mTOR activity; Inhibit the activity of at least one of mTORC1 or mTORC2; Inhibit the generation of at least one of mTORC1 or mTORC2; To cause at least one of mTORC1 or mTORC2 to dissociate; Degrade at least one of mTOR, mTORC1, or mTORC2; Repressing the transcription and / or translation of the mtor gene; and / or This can cause deletion and / or mutation of the mtor gene.

30. The use according to any one of claims 25-29, wherein, The drug or drug composition comprises at least one of the following: protein, polypeptide, nucleic acid, polynucleotide, polynucleotide, antibody or derivative thereof, organic synthetic compound, inorganic compound and / or natural product.

31. The use according to any one of claims 25-30, wherein, The drug or drug composition includes compounds that inhibit ATR activity and mTOR activity.

32. The use of any one of claims 25-31, wherein, The drug or drug composition includes ATR inhibitors and mTOR inhibitors.

33. The use according to claim 32, wherein, The ATR inhibitors include at least one of AZD6738, M-4344, M6620, BAY-1895344, RP-3500, M1774, ART0380, SKLB-197, AD1058, or their tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotopic derivatives, pharmaceutically acceptable salts, or solvates of pharmaceutically acceptable salts thereof; and / or The mTOR inhibitors include sirolimus, zotalimus, umirolimus, tesimolimus, gidalixetine, Paxalisib, SR-0379, Dactolisib, Oleanin, Olcorolimus, Everolimus, RM-006, desfolimus, SF-1126, KB02-SLF, LY-3023414, TAK-228, AZD2014, CC223, CC-115, RTB-101, Voxtalisib, ME-344, SCC-31, CC-214-2, DHW-208, FD-274, GNE-477, GNE-555, Panulisib, PF-04691503, Piplartine, PP30, PQR514, Torin2, Torkinib, X-387, PQR- 530, Torin1, AZD8055, KU-0063794, BGT-226, Bimiralisib, GDC-0349, Omilise, OSI-027, PF-0469 1502, PKI-179, XL-765, Apitolisib, DS-7423, GNE-493, MKC-1, P-2281, Palomids, PI-103, PWT- 33597, SN-32976, WYE-132, XL-388, PP242, PP30, XL388, WYE-354, WAY-600, WYE-687, AP23573, MLN0128 or at least one of the following: tautomer, stereoisomer, enantiomer, diastereomer, racemic mixture, solvate, isotopic derivative, pharmaceutically acceptable salt or solvate of a pharmaceutically acceptable salt thereof.

34. The use according to claim 31 or 32, wherein, The ATR inhibitor is selected from at least one of AZD6738, M-4344 or its tautomers, stereoisomers, enantiomers, diastereomers, racemic mixtures, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof; and / or The mTOR inhibitors include at least one of AZD2014, CC223 or its tautomers, stereoisomers, enantiomers, diastereomers, racemates, solvates, isotope derivatives, pharmaceutically acceptable salts or solvates of pharmaceutically acceptable salts thereof.

35. The use according to any one of claims 25-34, wherein, The dosage form of the drug or drug composition includes at least one of the following: solution, powder, granule, tablet, sugar-coated, capsule, granule, suspension, syrup, drops, and sublingual tablet.

36. The use of any one of claims 25-35, wherein, The pharmaceutical composition further includes pharmaceutically acceptable excipients, optionally including at least one of solubilizers, disintegrants, wetting agents, stabilizers, thickeners, diluents, buffers, and flavoring agents.

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