Pharmaceutical composition of KRAS G12C inhibitor and mitoxantrone and application thereof

Abstract

The invention relates to the field of antitumor drugs, in particular to a pharmaceutical composition of a KRAS G12C inhibitor and mitoxantrone and application of the pharmaceutical composition. The mitoxantrone hydrochloride and the KRAS G12C inhibitor are combined to play a synergistic effect, a DNA damage repair pathway is inhibited, a better inhibition effect is generated on tumor cells containing KRAS G12C mutation, the dosage required for achieving the same anti-tumor effect is greatly reduced, and the medication safety is improved.

Description

Technical Field

[0001] The present disclosure relates to the field of anti-tumor drugs, and in particular to a drug combination of a KRAS G12C inhibitor and mitoxantrone and use thereof in treating cancer. Background Art

[0002] RAS family GTPases (KRAS, NRAS, and HRAS) cycle between inactivation (GDP-bound) and activation (GTP-bound). This inactivation and activation is regulated by two types of factors: 1) guanine nucleotide exchange factors (GEFs, such as SOS proteins), which catalyze the binding of KRAS to GTP, thereby promoting KRAS activation; and 2) GTPase-activating proteins (GAPs), which promote the hydrolysis of KRAS-bound GTP to GDP, thereby inhibiting KRAS activation. Somatic KRAS mutations weaken the enzymatic activity of the protein, leading to the accumulation of GTP-bound, active KRAS, excessive activation of downstream signaling, and uncontrolled cell proliferation.

[0003] RAS mutation or amplification is one of the most common abnormalities in human cancer. KRAS mutations are most common, especially in solid tumors. Overall, about 17% of solid tumors have KRAS mutations, including about 90% of pancreatic cancer, about 50% of colon cancer, and about 25% of lung adenocarcinoma. G12C is one of the most common mutation types in the KRAS gene, accounting for about 14% of patients with non-small cell lung cancer, 4% of patients with colorectal cancer, and 3% of patients with pancreatic cancer. More than 100,000 people are diagnosed with this mutation each year worldwide. KRAS inhibitors inhibit the proliferation of tumor cells by blocking the activation of KRAS, thereby cutting off its downstream signaling pathways.

[0004] For many years, drug development targeting KRAS has been a hot topic of research. Existing attempts include targeting the KRAS protein itself and its post-translational modifications, membrane localization, protein-protein interactions, and downstream signaling pathways. However, most of these approaches have not been successful in clinical trials, and KRAS mutations have been considered "undruggable". In recent years, some new KRAS G12C inhibitors have been developed and entered clinical trials. Among them, Sotorasib (AMG510) and Adagrasib (MRTX849) are the most influential drugs. It has been reported in the literature that the response rate of Sotorasib for non-small cell lung cancer with KRAS G12C mutations is 32%, while the response rate of Adagrasib is 45%. Although both sotolacizumab and adagracib have been shown to be significantly effective against KRAS G12C tumors in clinical trials, their responsiveness in NSCLC is still insufficient compared with EGFR-TKI (about 70%) and ALK-TKI (about 80%). Therefore, how to sensitize this type of new KRAS G12C inhibitor in order to achieve better results has become a hot topic of research at home and abroad. Summary of the invention

[0005] The purpose of the present disclosure is to provide a technical solution to improve the anti-tumor efficacy of KRAS G12C inhibitors in view of the insufficient reactivity of KRAS G12C inhibitors.

[0006] To achieve the above technical objectives, in one aspect, the present disclosure provides a pharmaceutical composition or a drug kit, characterized in that it comprises a therapeutically effective amount of a KRAS G12C inhibitor and a therapeutically effective amount of mitoxantrone.

[0007] In some embodiments, the KRAS G12C inhibitor includes sotolacizumab and / or adagracib.

[0008] In some embodiments, the mitoxantrone is mitoxantrone hydrochloride.

[0009] In some embodiments, the pharmaceutical composition or kit comprises 3.125-100 nM of the KRAS G12C inhibitor and 3.125-100 nM of the mitoxantrone; in some preferred embodiments, the pharmaceutical composition or kit comprises 6.25-100 nM of the KRAS G12C inhibitor and 6.25-100 nM of the mitoxantrone; in some more preferred embodiments, the pharmaceutical composition or kit comprises 12.5-50 nM of the KRAS G12C inhibitor and 12.5-50 nM of the mitoxantrone.

[0010] In some embodiments, the molar ratio of the KRAS G12C inhibitor to the mitoxantrone is 0.03-32, for example, it can be 0.03, 0.06, 0.12, 0.25, 0.5, 1, 2, 4, 8, 16 or 32, preferably 0.06-16, more preferably 0.4-4, further preferably 0.5-4, and further preferably 1.

[0011] In some embodiments, the pharmaceutical composition or kit comprises 3.125 nM sotolacib and 100 nM mitoxantrone hydrochloride.

[0012] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM sotolacib and 100 nM mitoxantrone hydrochloride.

[0013] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM sotolacib and 50 nM mitoxantrone hydrochloride.

[0014] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM sotolacib and 50 nM mitoxantrone hydrochloride.

[0015] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM sotolacib and 25 nM mitoxantrone hydrochloride.

[0016] In some embodiments, the pharmaceutical composition or kit comprises 25 nM mitoxantrone hydrochloride and 25 nM sotolacib.

[0017] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM mitoxantrone hydrochloride and 25 nM sotolacib.

[0018] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM mitoxantrone hydrochloride and 50 nM sotolacib.

[0019] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM mitoxantrone hydrochloride and 50 nM sotolacib.

[0020] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM mitoxantrone hydrochloride and 100 nM sotolacib.

[0021] In some embodiments, the pharmaceutical composition or kit comprises 3.125 nM mitoxantrone hydrochloride and 100 nM sotolacib.

[0022] In some embodiments, the pharmaceutical composition or kit comprises 100 nM mitoxantrone hydrochloride and 100 nM sotolacib.

[0023] In some embodiments, the pharmaceutical composition or kit comprises 50 nM mitoxantrone hydrochloride and 50 nM sotolacib.

[0024] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM mitoxantrone hydrochloride and 12.5 nM sotolacib.

[0025] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM mitoxantrone hydrochloride and 6.25 nM sotolacib.

[0026] In some embodiments, the pharmaceutical composition or kit comprises 3.125 nM mitoxantrone hydrochloride and 3.125 nM sotolacib.

[0027] In some embodiments, the pharmaceutical composition or kit comprises 3.125 nM adagracib and 100 nM mitoxantrone hydrochloride.

[0028] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM adagracib and 100 nM mitoxantrone hydrochloride.

[0029] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM adagracib and 50 nM mitoxantrone hydrochloride.

[0030] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM adagracib and 50 nM mitoxantrone hydrochloride.

[0031] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM adagracib and 25 nM mitoxantrone hydrochloride.

[0032] In some embodiments, the pharmaceutical composition or kit comprises 25 nM mitoxantrone hydrochloride and 25 nM adagracib.

[0033] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM mitoxantrone hydrochloride and 25 nM adagracib.

[0034] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM mitoxantrone hydrochloride and 50 nM adagracib.

[0035] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM mitoxantrone hydrochloride and 50 nM adagracib.

[0036] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM mitoxantrone hydrochloride and 100 nM adagracib.

[0037] In some embodiments, the pharmaceutical composition or kit comprises 3.125 nM mitoxantrone hydrochloride and 100 nM adagracib.

[0038] In some embodiments, the pharmaceutical composition or kit comprises 100 nM mitoxantrone hydrochloride and 100 nM adagracib.

[0039] In some embodiments, the pharmaceutical composition or kit comprises 50 nM mitoxantrone hydrochloride and 50 nM adagracib.

[0040] In some embodiments, the pharmaceutical composition or kit comprises 12.5 nM mitoxantrone hydrochloride and 12.5 nM adagracib.

[0041] In some embodiments, the pharmaceutical composition or kit comprises 6.25 nM mitoxantrone hydrochloride and 6.25 nM adagracib.

[0042] In some embodiments, the pharmaceutical composition or kit comprises 3.125 nM mitoxantrone hydrochloride and 3.125 nM adagracib.

[0043] In another aspect, the present disclosure provides use of the above pharmaceutical composition in the preparation of a medicament or a drug kit for treating a disease or disease state containing a KRAS G12C mutation.

[0044] In yet another aspect, the present disclosure provides use of the above pharmaceutical composition in treating a disease or disease state containing a KRAS G12C mutation.

[0045] In another aspect, the present disclosure provides the use of a KRAS G12C inhibitor and mitoxantrone in the preparation of a medicament or kit for treating a disease or condition containing a KRAS G12C mutation.

[0046] In yet another aspect, the present disclosure provides for use of a KRAS G12C inhibitor and mitoxantrone in treating a disease or condition containing a KRAS G12C mutation.

[0047] According to any of the foregoing aspects, the disease or disease state comprises cancer; preferably, the cancer comprises solid tumors and / or hematological tumors; preferably, the cancer comprises non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, small intestine cancer, appendix cancer, colorectal cancer, cancer of unknown primary site, endometrial cancer, mixed cancer types, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplasia / myeloproliferative neoplasms, head and neck cancer, esophageal gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia or melanoma; more preferably, the cancer comprises non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colorectal cancer, bile duct cancer or pancreatic cancer.

[0048] In another aspect, the present disclosure provides use of mitoxantrone in the preparation of a medicament or a kit for improving the efficacy of a KRAS G12C inhibitor.

[0049] In yet another aspect, the present disclosure provides use of mitoxantrone in improving the efficacy of KRAS G12C inhibitors.

[0050] In another aspect, the present disclosure provides a drug or a kit for improving the efficacy of a KRAS G12C inhibitor, characterized in that it comprises mitoxantrone, wherein the mitoxantrone is administered at any time before, during or after administration of the KRAS G12C inhibitor.

[0051] In another aspect, the present disclosure provides a method of treating a disease or condition comprising a KRAS G12C mutation, comprising administering to an individual comprising a KRAS G12C mutation a therapeutically effective amount of a KRAS G12C inhibitor and a therapeutically effective amount of mitoxantrone.

[0052] In another aspect, the present disclosure provides a method for improving the efficacy of a KRAS G12C inhibitor, comprising administering mitoxantrone at any time before, during, or after administration of the KRAS G12C inhibitor.

[0053] According to any of the preceding aspects, the KRAS G12C inhibitor comprises sotolacizumab and / or adagracib.

[0054] According to any of the preceding aspects, the mitoxantrone is mitoxantrone hydrochloride.

[0055] According to any of the preceding aspects, the medicament or kit comprises 3.125-100 nM of the KRAS G12C inhibitor and 3.125-100 nM of the mitoxantrone; in some preferred embodiments, the medicament or kit comprises 6.25-100 nM of the KRAS G12C inhibitor and 6.25-100 nM of the mitoxantrone; in some more preferred embodiments, the medicament or kit comprises 12.5-50 nM of the KRAS G12C inhibitor and 12.5-50 nM of the mitoxantrone.

[0056] According to any of the preceding aspects, the molar ratio of the KRAS G12C inhibitor to the mitoxantrone is 0.03-32, for example, 0.03, 0.06, 0.12, 0.25, 0.5, 1, 2, 4, 8, 16 or 32, preferably 0.06-16, more preferably 0.4-4, more preferably 0.5-4, and more preferably 1.

[0057] In some embodiments, the medicament or kit comprises 3.125 nM sotolacib and 100 nM mitoxantrone hydrochloride.

[0058] In some embodiments, the medicament or kit comprises 6.25 nM sotolacib and 100 nM mitoxantrone hydrochloride.

[0059] In some embodiments, the medicament or kit comprises 6.25 nM sotolacib and 50 nM mitoxantrone hydrochloride.

[0060] In some embodiments, the medicament or kit comprises 12.5 nM sotolacib and 50 nM mitoxantrone hydrochloride.

[0061] In some embodiments, the medicament or kit comprises 12.5 nM sotolacib and 25 nM mitoxantrone hydrochloride.

[0062] In some embodiments, the medicament or kit comprises 25 nM mitoxantrone hydrochloride and 25 nM sotolacib.

[0063] In some embodiments, the medicament or kit comprises 12.5 nM mitoxantrone hydrochloride and 25 nM sotolacib.

[0064] In some embodiments, the medicament or kit comprises 12.5 nM mitoxantrone hydrochloride and 50 nM sotolacib.

[0065] In some embodiments, the medicament or kit comprises 6.25 nM mitoxantrone hydrochloride and 50 nM sotolacib.

[0066] In some embodiments, the medicament or kit comprises 6.25 nM mitoxantrone hydrochloride and 100 nM sotolacib.

[0067] In some embodiments, the medicament or kit comprises 3.125 nM mitoxantrone hydrochloride and 100 nM sotolacib.

[0068] In some embodiments, the medicament or kit comprises 100 nM mitoxantrone hydrochloride and 100 nM sotolacib.

[0069] In some embodiments, the medicament or kit comprises 50 nM mitoxantrone hydrochloride and 50 nM sotolacib.

[0070] In some embodiments, the medicament or kit comprises 12.5 nM mitoxantrone hydrochloride and 12.5 nM sotolacib.

[0071] In some embodiments, the medicament or kit comprises 6.25 nM mitoxantrone hydrochloride and 6.25 nM sotolacib.

[0072] In some embodiments, the medicament or kit comprises 3.125 nM mitoxantrone hydrochloride and 3.125 nM sotolacib.

[0073] In some embodiments, the medicament or kit comprises 3.125 nM adagracib and 100 nM mitoxantrone hydrochloride.

[0074] In some embodiments, the medicament or kit comprises 6.25 nM adagracib and 100 nM mitoxantrone hydrochloride.

[0075] In some embodiments, the medicament or kit comprises 6.25 nM adagracib and 50 nM mitoxantrone hydrochloride.

[0076] In some embodiments, the medicament or kit comprises 12.5 nM adagracib and 50 nM mitoxantrone hydrochloride.

[0077] In some embodiments, the medicament or kit comprises 12.5 nM adagracib and 25 nM mitoxantrone hydrochloride.

[0078] In some embodiments, the medicament or kit comprises 25 nM mitoxantrone hydrochloride and 25 nM adagracib.

[0079] In some embodiments, the medicament or kit comprises 12.5 nM mitoxantrone hydrochloride and 25 nM adagracib.

[0080] In some embodiments, the medicament or kit comprises 12.5 nM mitoxantrone hydrochloride and 50 nM adagracib.

[0081] In some embodiments, the medicament or kit comprises 6.25 nM mitoxantrone hydrochloride and 50 nM adagracib.

[0082] In some embodiments, the medicament or kit comprises 6.25 nM mitoxantrone hydrochloride and 100 nM adagracib.

[0083] In some embodiments, the medicament or kit comprises 3.125 nM mitoxantrone hydrochloride and 100 nM adagracib.

[0084] In some embodiments, the medicament or kit comprises 100 nM mitoxantrone hydrochloride and 100 nM adagracib.

[0085] In some embodiments, the medicament or kit comprises 50 nM mitoxantrone hydrochloride and 50 nM adagracib.

[0086] In some embodiments, the medicament or kit comprises 12.5 nM mitoxantrone hydrochloride and 12.5 nM adagracib.

[0087] In some embodiments, the medicament or kit comprises 6.25 nM mitoxantrone hydrochloride and 6.25 nM adagracib.

[0088] In some embodiments, the medicament or kit comprises 3.125 nM mitoxantrone hydrochloride and 3.125 nM adagracib.

[0089] The present invention combines the DNA topoisomerase II inhibitor mitoxantrone hydrochloride with the KRAS G12C inhibitor to exert a synergistic effect, inhibit the DNA damage repair pathway, produce a better inhibitory effect on tumor cells, significantly reduce the dose required to achieve the same anti-tumor effect, and improve the safety of medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0090] In order to more clearly illustrate the specific embodiments of the present disclosure or the technical solutions in the prior art, the drawings required for use in the specific embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings in the following description are some embodiments of the present disclosure. For ordinary technicians in this field, other drawings can also be obtained based on these drawings without creative work. The drawings herein are incorporated into the specification and constitute a part of the specification, showing embodiments that conform to the specification, and are used together with the specification to explain the principles of the specification.

[0091] Figure 1 The cell survival rate of the non-small cell lung cancer cell line NCI-H358 containing the KRAS G12C mutation treated with sotolacib, mitoxantrone hydrochloride and the combination of the two, as well as the effect value and combination index of the combination group are shown.

[0092] Figure 2 The cell survival rate of the lung cancer cell line NCI H2030 containing KRAS G12C mutation treated with sotolacib, mitoxantrone hydrochloride and their combination, as well as the effect value and combination index of the combination group are shown.

[0093] Figure 3 The cell survival rate of the lung cancer cell line Calu-1 containing KRAS G12C mutation treated with sotolacib, mitoxantrone hydrochloride and their combination, as well as the effect value and combination index of the combination group are shown.

[0094] Figure 4 The cell survival rate of the non-small cell lung cancer cell line NCI-H358 containing the KRAS G12C mutation treated with adagracib, mitoxantrone hydrochloride and the combination of the two, as well as the effect value and combination index of the combination group are shown. DETAILED DESCRIPTION

[0095] (I) Definition and Explanation

[0096] In order to make it easier to understand the present disclosure, some technical and scientific terms are specifically defined below. In the present disclosure, unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. At the same time, in order to better understand the present disclosure, the definition and explanation of related terms are provided below. It should be understood that the present disclosure is not limited to specific methods, reagents, compounds, compositions or biological systems, and of course the above can be changed. It should also be understood that the terms used in the present application are only for describing specific embodiments and are not intended to be limited.

[0097] As used in this specification and the appended claims, the singular forms "a," "an," "the," "said," and similar referents include plural referents unless the content clearly dictates otherwise.

[0098] As used herein, the conjunction term "and / or" between various elements is intended to include both the meanings of "and" and "or", for example, the phrase "A, B and / or C" is intended to cover each of the following aspects: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0099] As used herein, the terms "comprises," "comprising," "having," and "containing," and any variations thereof, are intended to cover a non-exclusive inclusion. The terms are intended to be open ended, to specify the presence of any stated features, elements, integers, steps, or components, but not to preclude the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" encompasses the more restrictive terms "consisting of" and "consisting essentially of."

[0100] Numerical ranges used herein should be understood to include all numbers within the range. For example, a range of 1 to 20 should be understood to include any number, combination of numbers, or subrange from the following group: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

[0101] As used herein, the term "about" means a range of ±20% of the value that follows. In some embodiments, the term "about" means a range of ±10% of the value that follows. In some embodiments, the term "about" means a range of ±5% of the value that follows.

[0102] In the description of this document, reference is made to “some embodiments”, “some implementation schemes” or “some implementation plans”, which describe a subset of all possible embodiments, but it can be understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0103] As used herein, the term "KRAS" refers to a Kirsten rat sarcoma viral oncogene homolog, a member of the small GTPase and Ras oncogene family. KRAS acts as a molecular switch cycle between an inactive (GDP-bound) state and an active (GTP-bound) state to convert upstream cellular signals received from multiple tyrosine kinases into downstream effectors that regulate a wide variety of processes including cell proliferation (see, e.g., Alamgeer et al., Current opinion in pharmacology, 2013, 13(3): 394-401, the contents of which are incorporated herein by reference). Single nucleotide substitutions leading to missense mutations at codons 12 and 13 of the KRAS primary amino acid sequence account for approximately 40% of these KRAS driver mutations in lung adenocarcinoma, with G12C transversion being the most common activating mutation (e.g., see Dogan et al., Clinical Cancer Research, 2012, 18(22):6169-6177, the contents of which are incorporated herein by reference), and KRAS gain-of-function mutations are found in approximately 30% of all human cancers. The KRAS G12C mutation is a specific submutation, prevalent in approximately 13% of lung adenocarcinomas, 3%-5% of colon adenocarcinomas, and a smaller fraction of other cancer types. The somatic KRAS G12C mutation reduces the enzymatic activity of the protein, leading to the accumulation of GTP-bound, active KRAS, excessive activation of downstream signaling, and thus uncontrolled cell proliferation.

[0104] As used herein, the term "KRAS G12C inhibitor" refers to compounds and / or compositions that can inhibit the proliferation of tumor cells by blocking the activation of KRAS G12C mutant protein and cutting off its downstream signaling pathway, such as sotolacib and adagraciib that are already on the market. Other KRAS G12C inhibitors in different preclinical or clinical trial stages should also be included in the scope of this disclosure, as long as they can inhibit the proliferation of tumor cells by blocking the activation of KRAS G12C mutant protein.

[0105] As used herein, the term "Sotorasib" (AMG510) includes Sotorasib and its pharmaceutically acceptable salts, which is a small molecule that specifically and irreversibly inhibits KRAS G12C mutant protein. The sotolacib is also known as AMG 510, with the chemical name 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(prop-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one, CAS number 2296729-00-3, which binds to the P2 pocket and nucleotide binding pocket adjacent to the mutant cysteine ​​at position 12 of KRAS. The inhibitor contains a thiol-reactive portion that covalently modifies the cysteine ​​residue and locks KRASG12C in an inactive guanosine diphosphate (GDP)-bound conformation. This blocks the interaction of KRAS with effectors, thereby preventing downstream signaling, including phosphorylation of extracellular signal-regulated kinase (ERK) (Cully and Downward, 2008; Ostrem et al., 2013; Simanshu et al., 2017, the contents of which are incorporated herein by reference).

[0106] As used herein, the term "Adagrasib" (MRTX849) includes Adagrasib and pharmaceutically acceptable salts thereof. The chemical name of Adagrasib is 2-[(2S)-4-[7-(8-chloro-1-naphthyl)-2-[[(2R)-1-methylpyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-1-(2-fluoroprop-2-enoyl)piperazin-2-yl]acetonitrile, with CAS number 2326521-71-3. It is the second KRAS G12C inhibitor launched in the world, used to treat locally advanced or metastatic non-small cell lung cancer and colorectal cancer carrying KRAS G12C mutations.

[0107] As used herein, the term "mitoxantrone" includes mitoxantrone and its pharmaceutically acceptable salts, for example, it can be one or more of mitoxantrone hydrochloride, mitoxantrone oxalate, mitoxantrone sulfate, mitoxantrone phosphate, mitoxantrone acetate, and mitoxantrone citrate, and the pharmaceutically acceptable salt of mitoxantrone is preferably mitoxantrone hydrochloride. The chemical name of mitoxantrone is 1,4-dihydroxy-5,8-di[[2-[(2-hydroxyethyl)amino]ethyl]amino]-9,10-anthraquinone, CAS No. 65271-80-9, which is an anthraquinone antibiotic antitumor drug, and its main mechanism of action is to insert into deoxyribonucleic acid through hydrogen bonding, causing cross-linking and breaking of DNA structure; it can interfere with RNA; mitoxantrone is also an effective inhibitor of topoisomerase II, which has a killing effect on both proliferative and non-proliferative cancer cells in the human body, and is a cell cycle non-specific drug. The main clinical application areas are malignant lymphoma, breast cancer and acute myeloid leukemia, lung cancer, melanoma, soft tissue sarcoma, multiple myeloma, liver cancer, colorectal cancer, kidney cancer, prostate cancer, endometrial cancer, testicular tumors, ovarian cancer and head and neck cancer, etc. At present, the common dosage forms of mitoxantrone on the market are mainly injections, including ordinary mitoxantrone hydrochloride injection and improved new dosage form of mitoxantrone hydrochloride liposome injection.

[0108] As used herein, the term "cancer" refers to a disease characterized by the uncontrolled growth of abnormal cells. Cancer cells can spread to other parts of the body locally or through the bloodstream and lymphatic system. Examples of various cancers are described herein, including but not limited to non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, prostate cancer, bladder cancer, liver cancer, skin cancer, glioma, breast cancer, melanoma, malignant glioma, rhabdomyosarcoma, ovarian cancer, astroglioma, Ewing's sarcoma, retinoblastoma, epithelial cell cancer, colon cancer, renal cancer, gastrointestinal stromal tumor, leukemia, lymphoma and nasopharyngeal carcinoma, etc. The terms "tumor" and "cancer" are used interchangeably in this article, for example, these two terms include solid and liquid, such as diffuse or circulating tumors. As used herein, the terms "cancer" or "tumor" include precancerous lesions and malignant cancers and tumors.

[0109] As used herein, the term "disease state" is or includes the likelihood of developing a clinically significant disease, such as the likelihood of developing a clinically significant disease within a particular time frame or natural life span, including the presence or likelihood of relapse in a subject, and / or whether a subject will benefit from treatment.

[0110] As used herein, the term "treatment" is intended to include prevention and treatment, for example, treatment of a disease comprising a KRAS G12C mutation includes prevention and / or treatment of a disease comprising a KRAS G12C mutation. Beneficial or desired results, including methods of clinical results, can be obtained by treatment. For the purposes of this application, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms caused by the disease, alleviating the extent of the disease, stabilizing the disease (e.g., preventing or delaying the deterioration of the disease), preventing or delaying the spread of the disease (e.g., metastasis), preventing or delaying the recurrence of the disease, delaying or slowing the progression of the disease, improving the disease state, providing relief (partial or complete) of the disease, reducing the dose of one or more other drugs required to treat the disease, delaying the progression of the disease, improving or improving the quality of life, increasing weight gain and / or prolonging survival. "Treatment" also includes a reduction in the pathological results of cancer (e.g., tumor volume). In the context of cancer, "treatment" includes any or all of the following: inhibiting the growth of cancer cells, inhibiting the replication of cancer cells, reducing overall tumor burden, and improving one or more symptoms associated with the disease.

[0111] As used herein, the term "effective amount" or "therapeutically effective amount" encompasses an amount sufficient to improve or prevent the symptoms or symptoms of a medical condition. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's overall health, the method, route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.

[0112] As used herein, the terms "subject", "individual" or "patient" are used interchangeably herein and refer to vertebrates, preferably mammals, more preferably humans. Mammals include, but are not limited to, mice, apes, humans, farm animals, competitive animals, and pets.

[0113] The amount of the drug administered may depend on the subject being treated, the subject's age, health, sex, and weight, the type of concurrent treatment (if any), the severity of the condition, the nature of the desired effect, the manner and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on arterial oxygen tension. However, the most preferred dosage may be adjusted for individual subjects, as will be understood by those skilled in the art and can be determined without undue experimentation. This generally includes adjusting the standard dosage (e.g., if the patient is of low weight, then reducing the dosage).

[0114] As used herein, the term "pharmaceutical composition" refers to a mixture of one or more components. The purpose of a pharmaceutical composition is to facilitate administration of a compound of the present disclosure to an organism. The pharmaceutical composition of the present disclosure may include one or more pharmaceutically acceptable salts, pharmaceutically acceptable carriers and / or adjuvants, such as antioxidants, preservatives, wetting agents, emulsifiers and dispersants, etc.

[0115] As used herein, the term "pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material, formulation adjuvant or carrier conventional in the art for use with a therapeutic agent, which together constitute a "pharmaceutical composition" for administration to an individual. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dosage and concentration employed and is compatible with the other ingredients of the formulation. A pharmaceutically acceptable carrier is suitable for the formulation employed. Suitable carriers are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, liposomes, polymer micelles or inorganic nanocarriers, etc.

[0116] The "pharmaceutical composition" of the present disclosure can also be administered to patients or subjects in need of such treatment in any suitable manner, such as oral, intravenous, parenteral, rectal, pulmonary or topical administration. When used for oral administration, the pharmaceutical composition can be prepared into oral preparations, such as oral solid preparations, such as tablets, capsules, pills, granules, etc.; or, oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When prepared into oral preparations, the pharmaceutical preparations may also contain suitable fillers, binders, disintegrants, lubricants, etc.

[0117] The pharmaceutical composition of the present invention can be prepared into any pharmaceutically acceptable dosage form for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and / or parenteral administration, for example, it can be formulated into tablets, lozenges, capsules, pills, solutions, suspensions, syrups, injections, suppositories, inhalants or sprays.

[0118] As used herein, the term "kit" refers to one or more components of a package, which can be packaged separately or placed in a container, such as a tube, bottle, vial, bag, blister, syringe or other suitable container device. The kit described herein may also include instructions for implementing the subject method using the components of the kit (e.g., instructions for preparing a pharmaceutical composition described herein and / or instructions for using a pharmaceutical composition). The instructions for implementing the subject method are usually recorded on a suitable recording medium. For example, the instructions can be printed on a substrate such as paper or plastic. Therefore, the instructions can be present in the kit in the form of a package insert, in the label of a container of the kit or its components (i.e., components associated with the package or small package). In some embodiments, the instructions exist as an electronic storage data file present in a suitable computer-readable storage medium (e.g., CD-ROM, disk, etc.). In other embodiments, the actual instructions are not present in the kit, but are provided for obtaining instructions from a remote source, such as through the Internet. An example of this embodiment is a kit comprising a website where the instructions can be viewed and / or from which the instructions can be downloaded.

[0119] As used herein, the term "synergistic effect" refers to a phenomenon in which the effects of two drugs used in combination are more effective than their individual effects, as opposed to antagonistic effects.

[0120] The "combination" or "combination use" described in the present disclosure is a mode of administration, which includes various situations in which two drugs are administered successively or simultaneously. The so-called "simultaneously" here means that the KRAS G12C inhibitor and mitoxantrone are administered in the same administration cycle, for example, the two drugs are administered within 2 days, or within 1 day. The so-called "sequential or sequential" administration includes the situation in which the KRAS G12C inhibitor and mitoxantrone are administered separately in different administration cycles. The "combined" administration method includes simultaneous administration, independent preparation and co-administration, or independent preparation and sequential administration. These administration methods all belong to the combined administration described in the present invention.

[0121] (II) Detailed technical solution

[0122] KRAS G12C inhibitors target KRAS G12C through a similar covalent binding mechanism, locking KRAS in an inactive state, thereby inhibiting the proliferation of tumor cells. However, due to the complexity of the KRAS downstream signaling pathway and the existence of multiple compensatory mechanisms, the response rate of KRAS G12C inhibitors in KRAS mutant tumors is insufficient. The inventors have conducted a large number of studies on the KRAS G12C inhibitors sotolacib and adaglaciib that have been on the market, and found that the DNA topoisomerase II inhibitor mitoxantrone can bind to the KRAS G12C inhibitors sotolacib or adaglaciib, inhibiting the DNA damage repair pathway, and exerting a synergistic effect with KRASG12C inhibitors, producing a better inhibitory effect on tumor cells.

[0123] In one aspect, the present disclosure provides a pharmaceutical composition or a drug kit, characterized in that it comprises a therapeutically effective amount of a KRASG12C inhibitor and a therapeutically effective amount of mitoxantrone. The mitoxantrone comprises mitoxantrone hydrochloride. The KRAS G12C inhibitor comprises sotolacib and / or adagracib.

[0124] In some embodiments, a pharmaceutical composition or kit is provided, which includes a therapeutically effective amount of sotolacib and a therapeutically effective amount of mitoxantrone. In some embodiments, sotolacib and mitoxantrone may be present in the same preparation, or may be separately prepared in the form of a combined package. When the above two components are prepared separately, the dosage forms may be the same or different. The dosage form may be any clinically acceptable dosage form, such as an injection dosage form, an oral dosage form, etc. The injection dosage form includes a liquid injection, an injection powder, an injection tablet, etc.; the oral dosage form includes a tablet, a capsule, an oral liquid preparation, etc.

[0125] In some embodiments, a pharmaceutical composition or kit is provided, which includes a therapeutically effective amount of adagrasib and a therapeutically effective amount of mitoxantrone. In some embodiments, adagrasib and mitoxantrone may be present in the same preparation, or they may be separately prepared in the form of a combined package. When the above two components are prepared separately, the dosage forms may be the same or different. The dosage form may be any dosage form that is clinically acceptable, such as an injection dosage form, an oral dosage form, and the like. The injection dosage form includes a liquid injection, an injection powder, an injection tablet, and the like; the oral dosage form includes a tablet, a capsule, an oral liquid preparation, and the like.

[0126] In another aspect, the present disclosure provides use of the above pharmaceutical composition in the preparation of a medicament or a drug kit for treating a disease or disease state containing a KRAS G12C mutation.

[0127] In yet another aspect, the present disclosure provides use of the above pharmaceutical composition in treating a disease or disease state containing a KRAS G12C mutation.

[0128] In another aspect, the present disclosure provides the use of a KRAS G12C inhibitor and mitoxantrone in the preparation of a medicament or kit for treating a disease or condition containing a KRAS G12C mutation.

[0129] In yet another aspect, the present disclosure provides for use of a KRAS G12C inhibitor and mitoxantrone in treating a disease or condition containing a KRAS G12C mutation.

[0130] In the above application, the disease or disease state includes cancer; preferably, the cancer includes solid tumors and / or hematological tumors; preferably, the cancer includes non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, small intestine cancer, appendix cancer, colorectal cancer, cancer of unknown primary site, endometrial cancer, mixed cancer types, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplasia / myeloproliferative neoplasms, head and neck cancer, esophageal gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia or melanoma; more preferably, the cancer includes non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colorectal cancer, bile duct cancer or pancreatic cancer.

[0131] In another aspect, the present disclosure provides use of mitoxantrone in the preparation of a medicament or a kit for improving the efficacy of a KRAS G12C inhibitor.

[0132] In yet another aspect, the present disclosure provides use of mitoxantrone in improving the efficacy of KRAS G12C inhibitors.

[0133] In yet another aspect, the present disclosure provides a method for improving the efficacy of a KRAS G12C inhibitor, comprising administering mitoxantrone at any time before, during, or after administration of the KRAS G12C inhibitor.

[0134] In another aspect, the present disclosure provides a drug or a kit for improving the efficacy of a KRAS G12C inhibitor, characterized in that it comprises mitoxantrone, wherein the mitoxantrone is administered at any time before, during or after administration of the KRAS G12C inhibitor.

[0135] According to any of the foregoing aspects, in some embodiments, the mitoxantrone is mitoxantrone hydrochloride. In some embodiments, the KRAS G12C inhibitor includes sotolanib and / or adagracib. In some embodiments, mitoxantrone is administered simultaneously with the KRAS G12C inhibitor. In some embodiments, mitoxantrone and the KRAS G12C inhibitor are administered successively in any order. In some embodiments, mitoxantrone and the KRAS G12C inhibitor have the same dosing cycle. In some embodiments, mitoxantrone and the KRAS G12C inhibitor have different dosing cycles.

[0136] On the other hand, the present disclosure provides a method for treating a disease or disease state containing a KRAS G12C mutation, comprising administering a therapeutically effective amount of a KRAS G12C inhibitor and a therapeutically effective amount of mitoxantrone to an individual containing a KRAS G12C mutation. The mitoxantrone is mitoxantrone hydrochloride. The KRAS G12C inhibitor includes sotolanib and / or adagracib. In some embodiments, mitoxantrone and a KRASG12C inhibitor are administered simultaneously to an individual containing a KRAS G12C mutation. In some embodiments, mitoxantrone and a KRAS G12C inhibitor are administered sequentially in any order to an individual containing a KRAS G12C mutation. In some embodiments, mitoxantrone and a KRAS G12C inhibitor are administered to an individual containing a KRAS G12C mutation in any suitable manner, such as oral, intravenous, parenteral, rectal, pulmonary or topical administration. The administration of mitoxantrone and a KRAS G12C inhibitor may be the same or different. The administration cycles of mitoxantrone and the KRAS G12C inhibitor may be the same or different.

[0137] Example

[0138] In order to make the purpose, technical scheme and advantages of the embodiments of the present disclosure clearer, the technical scheme in the embodiments of the present disclosure will be clearly and completely described below in conjunction with the drawings in the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all of the embodiments. In the absence of conflict, the following embodiments and the features in the embodiments can be combined with each other. The following examples are used to further illustrate the present disclosure, but should not be construed as limitations on the present disclosure. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present disclosure should all be equivalent replacement methods and are included in the protection scope of the present disclosure.

[0139] Example 1. Sotolacib and mitoxantrone hydrochloride have a synergistic effect in inhibiting cancer cells

[0140] The human non-small cell lung cancer cell line NCI-H358 with KRAS-G12C mutation was selected (purchased from Wuhan Punosai, catalog number CL-0400), and the cells were inoculated in RPMI1640 cell culture medium containing 10% fetal bovine serum and 1% penicillin / streptomycin double antibody, and placed in a cell culture incubator at 37°C with 5% CO2. The medium was changed every 2-3 days, and the cells were digested with 0.25% trypsin, passaged and collected. The cells in the logarithmic growth phase were prepared with RPMI1640 cell culture medium containing 10% fetal bovine serum and 1% penicillin / streptomycin double antibody to make 5×10 4 / mL concentration of cell suspension, at a concentration of 5000 cells / 100μL per well, added to a 96-well cell culture plate. After 24 hours of culture, the original culture medium was aspirated, and 100μL of culture medium containing different concentrations of the test substance was added to each well. Six parallel wells were set for each concentration. After 72 hours of culture, the original culture medium was aspirated. 100μL of freshly prepared RPMI1640 cell culture medium containing 10% CCK-8 solution was added to each well, and the supernatant was discarded after culturing at 37℃ for 1 hour. After wiping the bottom of the 96-well plate, the absorbance (OD value) was detected with an enzyme marker, and the detection wavelength was 450nm. The concentrations of the test samples were set as follows: 100nM, 50nM, 25nM, 12.5nM, 6.25nM, and 3.125nM for sotolacib, and 100nM, 50nM, 25nM, 12.5nM, 6.25nM, and 3.125nM for mitoxantrone hydrochloride, each with 6 concentrations. The concentrations of sotolacib and mitoxantrone hydrochloride in the combination group were the same, 100nM, 50nM, 25nM, 12.5nM, 6.25nM, and 3.125nM, respectively, and a blank control was set. Before use, the drug was dissolved in cell culture medium.

[0141] Data processing: Cell survival rate = 1-(control group OD value - drug group OD value) / control group OD value × 100%. GraphPad Prism 9.5.1 software was used to draw the graph, and the tumor cell growth curve was fitted with the [Inhibitor] vs. normalized response--Variable slope equation in the software. CompuSyn1.0 was used to calculate the combination index (CI) for evaluating the effect of the combination of two drugs. The CI value is an important indicator for evaluating the synergistic effect of the combination of two drugs. This value represents the ratio of the concentration required to achieve a certain inhibition rate X% after the combination of the two drugs to the concentration required to achieve the same inhibition rate X% when the two drugs are used alone. Therefore, 0<CI<1 indicates that the two drugs have a synergistic effect, CI value = 1 indicates that the two drugs have an additive effect, and CI value>1 indicates that the two drugs have an antagonistic effect. It is generally believed that the smaller the CI value, the stronger the synergistic effect of the two drugs. Fa (Fraction affected) is the impact fraction, that is, the quantitative value of the drug effect, also known as the effect value, which is expressed by the cell inhibition rate in this article.

[0142] The specific results are shown in Table 1 and Figure 1 At all concentrations, sotolacib and mitoxantrone hydrochloride have strong synergistic effects. After the two drugs are combined, only 30% of the original single drug dose is needed to achieve the same anti-tumor effect, which improves the safety of medication and enhances the anti-tumor effect.

[0143] Table 1. Survival rate of NCI H358 cells treated with different concentrations of sotolacib and mitoxantrone hydrochloride and their combination, as well as the effect value (Fa) and combination index (CI) of the combination group

[0144]

[0145] Example 2. Sotolacib and mitoxantrone hydrochloride have a synergistic effect in inhibiting cancer cells with KRAS G12C mutation

[0146] The experimental method and data processing were the same as in Example 1, except that the NCI H358 cell line was replaced with the KRAS G12C mutant lung cancer cell line NCI H2030 or Calu-1. The specific results are shown in Figure 2 and Figure 3 As shown, sotolacib and mitoxantrone hydrochloride exhibited stronger synergistic inhibitory effects on the cancer cell lines NCI H2030 or Calu-1 with KRAS G12C at low concentrations, which can significantly reduce the dosage of the drugs.

[0147] Example 3. Adaglasib and mitoxantrone hydrochloride have a synergistic effect in inhibiting cancer cells with KRAS G12C mutation

[0148] The experimental method and data processing were the same as in Example 1, except that the KRAS G12C inhibitor sotolacizumab was replaced with adagracib. The specific results are as follows Figure 4 As shown, adagracib and mitoxantrone hydrochloride showed strong synergistic effects at all drug concentrations, which can reduce the drug dosage and enhance the cancer cell inhibition effect.

[0149] Due to the availability of materials, the above examples illustrate the synergistic effects of the available KRAS G12C inhibitors sotolacib and adagracib with mitoxantrone, but those skilled in the art can understand that based on the same principle of action on the same target, other KRAS G12C inhibitors can have similar synergistic effects with mitoxantrone. The purpose of selecting and describing exemplary embodiments in this disclosure is to explain the specific principles of the present invention and its practical application. Those of ordinary skill in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of this disclosure.

Claims

1. A pharmaceutical composition or a medicine kit, characterized in that: The invention comprises a therapeutically effective amount of a KRAS G12C inhibitor and a therapeutically effective amount of mitoxantrone.

2. The pharmaceutical composition or kit according to claim 1, wherein the KRAS G12C inhibitor comprises sotolacizumab and / or adagracib.

3. The pharmaceutical composition or kit according to claim 1 or 2, wherein: The pharmaceutical composition or kit comprises 3.125-100 nM of the KRAS G12C inhibitor and 3.125-100 nM of mitoxantrone; Preferably, the pharmaceutical composition or kit comprises 6.25-100 nM of the KRAS G12C inhibitor and 6.25-100 nM of mitoxantrone; More preferably, the pharmaceutical composition or kit comprises 12.5-50 nM of the KRAS G12C inhibitor and 12.5-50 nM of mitoxantrone.

4. The pharmaceutical composition or kit according to claim 1 or 2, wherein: The molar ratio of the KRAS G12C inhibitor to the mitoxantrone is 0.03-32, preferably 0.06-16, more preferably 0.4-4, and more preferably 1.

5. The pharmaceutical composition or kit according to any one of claims 1 to 4, wherein the mitoxantrone is mitoxantrone hydrochloride.

6. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a medicament or a drug kit for treating a disease or disease state containing a KRAS G12C mutation.

7. Use of a KRAS G12C inhibitor and mitoxantrone in the preparation of a medicament or a kit for treating a disease or disease state containing a KRAS G12C mutation.

8. The use according to claim 6 or 7, wherein the disease or disease state comprises cancer; Preferably, the cancer comprises solid tumors and / or hematological tumors; Preferably, the cancer comprises non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, small intestine cancer, appendix cancer, colorectal cancer, cancer of unknown primary site, endometrial cancer, mixed cancer types, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplasia / myeloproliferative neoplasm, head and neck cancer, esophageal gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia or melanoma; Preferably, the cancer includes non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colorectal cancer, bile duct cancer or pancreatic cancer.

9. Use of mitoxantrone in the preparation of a medicament or a medicine kit for improving the efficacy of a KRAS G12C inhibitor.

10. The use according to any one of claims 7 to 9, wherein the mitoxantrone is mitoxantrone hydrochloride; The KRAS G12C inhibitors include sotolacizumab and / or adagracib.

11. The use according to any one of claims 7, 8 and 10, wherein: The drug or drug kit comprises 3.125-100 nM of the KRAS G12C inhibitor and 3.125-100 nM of mitoxantrone; Preferably, the drug or drug kit comprises 6.25-100 nM of the KRAS G12C inhibitor and 6.25-100 nM of mitoxantrone; More preferably, the medicament or kit comprises 12.5-50 nM of the KRAS G12C inhibitor and 12.5-50 nM of mitoxantrone.

12. The use according to any one of claims 7 to 11, wherein: The molar ratio of the KRAS G12C inhibitor to the mitoxantrone is 0.03-32, preferably 0.06-16, more preferably 0.4-4, and more preferably 1.

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