PIM kinase inhibitors in combination with KRAS inhibitors

Abstract

The present invention relates to a combination of a PIM kinase inhibitor and a KRAS inhibitor for use in treating cancer in a subject in need of such treatment. Compositions or kits containing the same are also provided. The present invention also relates to a PIM kinase inhibitor for use in treating cancer with a KRAS mutation in a human subject in need of such treatment.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a combination therapy comprising a PIM kinase inhibitor and a KRAS inhibitor for the treatment of cancer. The present invention also relates to a pharmaceutical composition or kit comprising a PIM kinase inhibitor and a KRAS inhibitor for the treatment of cancer. The present invention also relates to a PIM kinase inhibitor for use in the treatment of cancers with KRAS mutations. [Background technology]

[0002] background KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer (CRC), gallbladder cancer, thyroid cancer, and bile duct cancer. Among KRAS mutations, the G12 codon (81%) is the most frequently mutated, followed by G13 (14%) and Q61 (2%). KRAS mutations are the most common RAS mutation in pancreatic cancer (88%), followed by colon adenocarcinoma (50%), rectal adenocarcinoma (50%), lung adenocarcinoma (32%), small intestine adenocarcinoma (26%), cholangiocarcinoma (23%), plasma cell myeloma (18%), gallbladder cancer (16%), and anaplastic thyroid cancer (8.6%) (Kwan et al. J Exp Clin Cancer Res (2022) 41:27). Therefore, there is strong interest in drugs that block proliferative signaling induced by oncogenic KRAS variants.

[0003] KRAS was considered an undruggable target for decades, but in 2013, researchers discovered a KRAS molecule that was only exposed in its GDP-bound form, ultimately resulting in a direct drug-binding site. G12CA hidden pocket adjacent to the mutant cysteine ​​in the protein has been identified. Various attempts have been made to develop KRAS inhibitors. In May 2021, AMG510 (sotorasib) became the first FDA-approved therapeutic agent directly targeting KRAS-mutated tumors. In June 2021, MRTX849 (adagrasib) received Breakthrough Therapy Designation by the FDA. Additional KRAS inhibitors such as ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), MRTX1257, LY3499446, LY3537982, BI1823911, GDC-6036, RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MRTX1133, BI1701963, and BAY-293 are currently in development.

[0004] KRAS G12C Despite the clinical benefit observed in many patients treated with the inhibitors adagrasib or sotorasib, acquired resistance to monotherapy eventually develops in the majority of patients. Patients with KRAS mutant tumors have a significantly worse outcome and poor prognosis. G12C WO2020106647A2 discloses a KRAS inhibitor in combination with carboplatin, an anti-PD-1 inhibitor, a MEK inhibitor, an EGFR inhibitor, a TOR inhibitor, an SHP2 inhibitor, a PI3K inhibitor or an AKT inhibitor. G12C Jane de Lartigue also discloses KRAS inhibitors in combination with pan-ERBB inhibitors, CDK4 / 6 inhibitors, and SOS1 / pan-KRAS inhibitors. G12C Inhibitors are disclosed (Jane de Lartigue. OncologyLive, Vol. 23 / No. 1, Volume 23, Issue 01. table).

[0005] Furthermore, several MAPK pathway-targeting therapeutic agents are contraindicated for the treatment of KRAS mutant tumors due to a lack of clinical efficacy. Furthermore, non-tumor or non-mutant selective therapeutic agents may introduce on-target toxicity due to MAPK signaling inhibition in normal cells. This limits the use of such agents in combination with standard therapy or immunotherapy. Therefore, there is a significant unmet need for the development of tumor-selective therapeutic agents that do not introduce damage to normal cells. To date, there have been few combination therapies using PIM inhibitors and KRAS inhibitors, particularly PIM inhibitors and KRAS inhibitors. G12C inhibitor, KRAS G12D inhibitors or KRAS G12V There have been no studies of inhibitor combinations. Summary of the Invention

[0006] Summary of the Invention To overcome resistance to long-term KRAS inhibitor treatment, the present invention provides a combination therapy comprising a PIM kinase inhibitor (PIMi) and a KRAS inhibitor. It has been found that resistance to KRAS inhibitors, particularly acquired resistance, can be reversed by co-treatment with a PIM kinase inhibitor. Furthermore, the combination of a PIM kinase inhibitor and a KRAS inhibitor demonstrated superior efficacy compared to either single-agent treatment and demonstrated synergy against KRAS mutant cancers. Furthermore, it has been found that the PIM inhibitor GDC-0570 can be used as a single active agent in various KRAS mutant cancers.

[0007] Based on these findings, the present invention relates to the following aspects.

[0008] In some embodiments, the present invention provides a method for treating cancer in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a PIM kinase inhibitor and a KRAS inhibitor.In some embodiments, the cancer is a KRAS mutant cancer or a KRAS inhibitor-resistant cancer.

[0009] In other embodiments, the present invention provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the treatment of cancer, such as a KRAS mutant cancer or a KRAS inhibitor resistant cancer.

[0010] In another aspect, the present invention provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the manufacture of a medicament for use in the treatment of cancer, such as a KRAS mutant cancer or a KRAS inhibitor resistant cancer.

[0011] In another embodiment, the present invention provides a pharmaceutical composition comprising a PIM kinase inhibitor and a KRAS inhibitor. Preferably, the pharmaceutical composition is for use in treating cancer, such as a KRAS mutant cancer or a KRAS inhibitor-resistant cancer.

[0012] In another aspect, the present invention provides a kit comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable excipient.

[0013] In yet another aspect, the present invention provides a method of treating a cancer harboring a KRAS mutation in a human subject in need thereof, comprising administering a therapeutically effective amount of N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide (GDC-0570) or a pharmaceutically acceptable salt thereof. GDC-0570 may be used as a single active agent in a variety of KRAS mutant cancers. [Brief explanation of the drawings]

[0014] [Figure 1] Shows the effect of GDC-0570 and sotorasib on tumor volume in the KRASG12C mutant CRC PDX model CRC024.

[0015] [Figure 2]Shows the effect of GDC-0570 and sotorasib on tumor volume in the KRASG12C mutant NSCLC PDX model LUN055.

[0016] [Figure 3] Shows the effect of GDC-0570 and sotorasib on tumor volume in the KRASG12C mutant NSCLC PDX model LUN156.

[0017] [Figure 4] Shows the effect of GDC-0570 and sotorasib on tumor volume in the KRASG12C mutant NSCLC PDX model LUN2156-44.

[0018] [Figure 5] Shows the effect of GDC-0570 and GDC-6036 on tumor volume in the KRASG12C mutant CRC PDX model CRC022.

[0019] [Figure 6] Shows the effect of GDC-0570 on tumor volume in the KRASG12C mutant NSCLC PDX model LUN156.

[0020] [Figure 7] Shows the effect of GDC-0570 on tumor volume in KRASG12D mutant NSCLC PDX model LUN#137.

[0021] [Figure 8] Shows the effect of GDC-0570 and cobimetinib on tumor volume in the KRASG12D mutant pancreatic PDX model PAN092.

[0022] [Figure 9] 1 shows the effect of GDC-0570 and MRTX1133 on tumor volume in the KRASG12D pancreatic cancer model PAN092.

[0023] [Figure 10] Shows the effect of GDC-0570 and cobimetinib on tumor volume in KRASG12V mutant CRC PDX models. DETAILED DESCRIPTION OF THE INVENTION

[0024] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0025] The terms "comprise," "include," "comprises," "including," "comprises," and "comprises" are intended to specify the presence of a stated feature, integer, component, or step, but do not exclude the presence or addition of one or more other features, integers, components, steps, or groups thereof.

[0026] As used herein, the terms "a," "an," and "the" include both singular and plural referents unless the context clearly dictates otherwise.

[0027] The term "optionally" or "optionally" means that the subsequently described event, circumstance, or substituent may or may not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur.

[0028] The term "about" or "approximately" generally means within 5% or, more preferably, within 1% of a given value or range.

[0029] The recitation of numerical ranges by endpoints includes all values ​​and fractions subsumed within the range, as well as the recited endpoints.

[0030] The terms "treatment," "treating," and "treating" refer to therapeutic treatment in a subject with cancer, where the goal is to prevent or slow (reduce) unwanted physiological changes or disorders, such as the growth, progression, or spread of cancer. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of disease, stabilization (i.e., not worsening) of disease, delay or slowing of disease progression, improvement or palliation and remission (whether partial or complete) of the disease state. "Treatment" can also mean prolonging survival compared to life expectancy without treatment.

[0031] The term "therapeutically effective amount" refers to the amount of a combination of a PIM kinase inhibitor and a KRAS inhibitor that (i) treats cancer, (ii) attenuates, ameliorates, or eliminates one or more symptoms of cancer, and / or (iii) prevents or delays the onset of one or more symptoms of cancer, where the amount of the combination shows improvement in (i), (ii), or (iii) compared to monotherapy. A therapeutically effective amount of the combination may be capable of reducing the number of cancer cells; reducing tumor size; preventing (i.e., slowing down to some extent and preferably stopping) cancer cell invasion into peripheral organs; preventing (i.e., slowing down to some extent and preferably stopping) tumor metastasis; preventing tumor growth to some extent; and / or alternatively alleviating to some extent one or more symptoms associated with cancer. To the extent that the combination can prevent and / or kill existing cancer cells, it may be cytostatic and / or cytotoxic. For cancer treatment, efficacy can be measured, for example, by assessing the time to progression (TTP) and / or determining the response rate (RR).

[0032] As used herein, the term "synergistic" refers to a therapeutic combination that is more effective than the additive effects of two or more single agents. Determination of synergistic interactions between a PIM kinase inhibitor and a KRAS inhibitor can be based on results from the assays described herein. Combination therapy can provide "synergistic" effects, i.e., the effect achieved when multiple active ingredients are used together is greater than the sum of the effects resulting from using the compounds separately. A synergistic effect can be achieved, for example, by multiple active ingredients (1) co-formulated into a unit dosage form and administered or delivered simultaneously in combination; (2) delivered alternately or in parallel as separate formulations; or (3) by some other regimen. When delivered in alternation therapy, a synergistic effect can be achieved when the multiple compounds are administered or delivered sequentially, for example, by separate injections in separate syringes or by separate oral administration. Generally, during alternation therapy, the effective dosage of each active ingredient is administered sequentially, i.e., consecutively, while in combination therapy, the effective dosage of two or more active ingredients is administered together.Synergism can be evaluated by tumor growth inhibition.Specifically, the tendency of tumor volume growth is suppressed, and preferably tumor volume is significantly reduced or tumor is in complete regression.

[0033] "Resistant cancer" or "refractory cancer" are used interchangeably herein and refer to cancers that are unresponsive or hyporesponsive to therapeutic treatment. Resistant cancers can have intrinsic resistance, acquired resistance, or adaptive resistance. "Intrinsic resistance" or "primary resistance" refers to a lack of tumor response to initial treatment. "Acquired resistance" refers to tumors that initially respond to treatment and later recur. "Adaptive resistance" refers to resistance induced by alterations in upstream, downstream, or parallel pathway components in KRAS mutant cancers, inevitably resulting in a lack of efficacy and leading to the recurrence and progression of these tumors. For example, "KRAS-resistant cancer" can include cancers with acquired resistance to one or more KRAS inhibitors due to prior treatment with one or more KRAS inhibitors, or cancers with intrinsic resistance to one or more KRAS inhibitors, such as cancers with activated PIM kinases.

[0034] Resistant cancers may be resistant from the start of treatment or may become resistant during treatment.

[0035] As used herein, the term "sensitive cancer" refers to a cancer that responds to a drug treatment without progressing. For example, a sensitive cancer is a sotrasil-sensitive KRAS G12C Mutant colorectal cancer or sotorasib-sensitive KRAS G12C These cancer PDX models are mutant NSCLC cancers. Administration of 100 mg / kg sotorasib results in >100% TGI.

[0036] As used herein, the term "low-sensitivity cancer" refers to cancer that progresses with a certain drug treatment. For example, a low-sensitivity cancer is a cancer that progresses with a sotrasib-low-sensitivity KRAS G12C This is a mutant NSCLC cancer. Administration of 100mg / kg sotorasib to this cancer PDX model resulted in <100% TGI.

[0037] The term "combination" refers to simultaneous, separate or sequential administration. When administration is sequential or separate, the delay in administration of the second component should not be such as to eliminate the beneficial effect of the combination.

[0038] As used herein, a subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is an adult subject. In some embodiments, the subject is a male human subject. In some embodiments, the subject is a female human subject. "Subject" and "patient" and "individual" are also used interchangeably herein.

[0039] The term "pharmaceutically acceptable" indicates that a substance or composition must be chemically and / or toxicologically compatible with other ingredients included in the formulation and / or the mammal being treated therewith.

[0040] The term " pharmaceutically acceptable salt " refers to the pharmaceutically acceptable organic or inorganic salt of a compound. Examples of salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, superphosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, acid tartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate (mesylate), ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Pharmaceutically acceptable salts may involve the inclusion of other molecules, such as acetate ions, succinate ions, or other counter ions. The counter ion may be any organic or inorganic moiety that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. If it is part of multiple charged atoms, the pharmaceutically acceptable salt may have multiple counter ions. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counter ions. If the compound is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, by treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, or an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid such as glucuronic acid or galacturonic acid, an alpha hydroxy acid such as citric acid or tartaric acid, an amino acid such as aspartic acid or glutamic acid, an aromatic acid such as benzoic acid or cinnamic acid, a sulfonic acid such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.Acids generally considered suitable for forming pharmaceutically useful or acceptable salt forms from basic pharmaceutical compounds are described, for example, in P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66(1) 1 19; P. Gould, International J. of Pharmaceutics (1986) 33 201 217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; Remington's Pharmaceutical Sciences, 18th ed., (1995) Mack Publishing Co., Easton PA; and The Orange Book (Food & Drug Administration, Washington, DC on their website). If the compound is an acid, the desired pharmaceutically acceptable salt can be prepared by any suitable method, for example, by treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary), an alkali metal hydroxide, or an alkaline earth metal hydroxide. Illustrative examples of suitable salts include, but are not limited to, organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines such as piperidine, morpholine, and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

[0041] All publications, published patent documents, and patent applications cited herein are herein incorporated by reference to the same extent as if each individual publication, published patent document, or patent application was specifically and individually indicated to be incorporated by reference.

[0042] Detailed Description 1. Exemplary PIM Kinase Inhibitors and KRAS Inhibitors The present invention generally relates to the combination of a PIM kinase inhibitor as described herein with a KRAS inhibitor, for use, for example, in the treatment of cancer.

[0043] In some embodiments, the PIM kinase inhibitor is a PIM-1 kinase inhibitor, a PIM-2 kinase inhibitor, or a PIM-3 inhibitor. In some embodiments, the PIM kinase inhibitor is a pan-PIM kinase inhibitor that exhibits potent activity against PIM-1, PIM-2, and / or PIM-3 inhibitors. Exemplary PIM kinase inhibitors include, but are not limited to, AZD1208, LGH447, and compounds disclosed in WO2014048939, US20110059961, or US20130079321 (e.g., GDC-0570, GNE-1571, GNE-5775, GDC-0339, and GNE-5652), and pharmaceutically acceptable salts thereof. [ka]

[0044] In certain embodiments, the PIM kinase inhibitor is selected from the group consisting of GDC-0570, GNE-1571, GNE-5775, GDC-0339, GNE-5652, and pharmaceutically acceptable salts thereof.

[0045] In certain embodiments, the PIM kinase inhibitor is GDC-0570, also known as N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide, or a pharmaceutically acceptable salt thereof. GDC-0570 is compound 321 in WO2014048939.

[0046] In certain embodiments, the PIM kinase inhibitor is GNE-1571, also known as N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2-fluorophenyl)thiazole-4-carboxamide, or a pharmaceutically acceptable salt thereof. GNE-1571 is compound 322 in WO2014048939.

[0047] In some embodiments, the PIM kinase inhibitor is GNE-5775, also known as N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(3-methylpyridin-2-yl)thiazole-4-carboxamide, or a pharmaceutically acceptable salt thereof. GNE-5775 is compound 231 in WO2014048939.

[0048] In certain embodiments, the PIM kinase inhibitor is GDC-0339, also known as 5-amino-N-(5-((4R,5R)-4-amino-5-fluoroazepan-1-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide, or a pharmaceutically acceptable salt thereof. GDC-0339 is the compound of Example 139 in US20130079321.

[0049] In some embodiments, the PIM kinase inhibitor is GNE-5652, also known as (S)-5-amino-N-(4-(3-aminopiperidin-1-yl)pyridin-3-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide, or a pharmaceutically acceptable salt thereof. GNE-5652 is the compound of Example 3 in US20110059961.

[0050] In certain embodiments, the KRAS inhibitor inhibits KRAS G12C inhibitor, KRAS G12V inhibitor, KRAS G12D inhibitor, KRAS G13C inhibitor, KRASG13D inhibitor, KRAS Q61H inhibitor, KRAS Q61L inhibitor, KRAS Q61R inhibitor, KRAS K117N inhibitor, pan-KRAS inhibitor, KRAS-SOS1 interaction inhibitor or KRAS signaling inhibitor, and pharmaceutically acceptable salts thereof.

[0051] In one embodiment, KRAS G12C Inhibitors include sotorasib (AMG510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and pharmaceutically acceptable salts thereof. G12C The structures of the inhibitors are shown below. [ka]

[0052] In one embodiment, KRAS G12CInhibitors include WO2014152588, WO2015054572, WO2016049524, WO2016164675, WO2016168540, WO2017015562, WO2017058915, WO2017058807, WO2017058792, WO2017058902, WO2017087528, WO2017201161, WO2018064510, WO2018068017, WO2018119183, and WO201814 0600, WO2018140512, WO2018143315, WO2018206539, WO2018217651, WO2018218070, WO2019051291, WO2019099524, WO201911 0751, WO2019137985, WO2019141250, CN111377918CN112159405, CN112574199, WO2021155716, WO2021197499, WO2021249563 As disclosed in WO2022028346, WO2022037560, WO2022068921, WO2022111521 and WO2022135591.

[0053] In one embodiment, KRAS G12D The inhibitor is selected from the group consisting of MRTX1133, JAB-22000, RMC-9805 (RM-036) and pharmaceutically acceptable salts thereof. [ka]

[0054] In one embodiment, KRAS G13C The inhibitor is selected from the group consisting of RMC-8839 and pharmaceutically acceptable salts thereof.

[0055] In one embodiment, KRAS G12V The inhibitor is selected from JAB-23000 and pharmaceutically acceptable salts thereof.

[0056] In certain embodiments, the pan-KRAS inhibitor is selected from the group consisting of RMC-6236, BBP-454, and pharmaceutically acceptable salts thereof.

[0057] In one embodiment, the KRAS signaling inhibitor is a MEK inhibitor that inhibits mitogen-activated protein kinase 1 (MAP2K1 or MEK1) and central components of the RAS / RAF / MEK / ERK signaling pathway. In one embodiment, the KRAS signaling inhibitor is the MEK inhibitor cobimetinib. Cobimetinib inhibits KRAS G12D , KRAS G12V Inhibits all oncogenic KRAS downstream signaling, such as [ka]

[0058] In some embodiments, the KRAS-SOS1 interaction inhibitor is selected from the group consisting of BI1701963, BAY-293, RMC-5845, BI-3406, SDGR5, and pharmaceutically acceptable salts thereof. In some embodiments, the KRAS-SOS1 interaction inhibitor is disclosed in WO2018115380, WO2019122129, WO2018172250, WO2016077793, and WO2022017339.

[0059] PIM kinase inhibitors and KRAS inhibitors can be present as isotopically labeled compounds that are identical to those described herein, except that one or more atoms are replaced with atoms having atomic masses or mass numbers different from the atomic masses or mass numbers that are usually found in nature.All isotopes of any specified specific atoms or elements are contemplated within the scope of the compounds of the present invention and their applications.Exemplary isotopes that can be incorporated into compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine, for example. 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N,15 O. 17 O. 18 O. 32 P, 33 P, 35 S, 18 F, 36 Cl 123 I and 125 I. Certain isotopically labeled compounds (e.g., 3 H and 14 C) are useful for compound and / or substrate tissue distribution assays. 3 H) and carbon-14 ( 14 C) isotopes are useful because of their ease of production and detectability. 2 Substitution with heavier isotopes, such as 3H, may offer certain therapeutic advantages (e.g., increased in vivo half-life or reduced dosage requirements) due to greater metabolic stability, and therefore may be preferable in certain situations. 15 O. 13 N, 11 C and 18 Positron-emitting isotopes such as F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0060] In certain embodiments, the PIM kinase inhibitor is selected from GDC-0570, GNE-1571, GNE-5775, GDC-0339, and GNE-5652, and pharmaceutically acceptable salts thereof, and is a KRAS G12C The inhibitor is selected from sotorasib (AMG510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and pharmaceutically acceptable salts thereof.

[0061] 2. Cancer treatment In another aspect, the present invention provides a method of treating cancer, such as a KRAS mutant cancer or a KRAS inhibitor-resistant cancer, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a PIM kinase inhibitor and a therapeutically effective amount of a KRAS inhibitor.

[0062] In other embodiments, the present invention provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the treatment of cancer, such as a KRAS mutant cancer or a KRAS inhibitor resistant cancer.

[0063] In another aspect, the present invention provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the manufacture of a medicament for use in the treatment of cancer, such as a KRAS mutant cancer or a KRAS inhibitor resistant cancer.

[0064] The terms "tumor" and "cancer" are used interchangeably herein and refer to a physiological condition in mammals that is typically characterized by uncontrolled cell growth. In some embodiments, the cancer has a KRAS p.G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutation, or a combination thereof. In some embodiments, the cancer has a KRAS p.G12C or G12D mutation, or a combination thereof.

[0065] In some embodiments, cancers include, but are not limited to, lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, small intestine cancer, colon adenocarcinoma, rectal adenocarcinoma, small intestine adenocarcinoma, lung adenocarcinoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, plasma cell myeloma, gallbladder cancer, anaplastic thyroid cancer, ampullary carcinoma, cervical cancer, gastrointestinal neuroendocrine tumors, uterine endometrioid carcinoma, germ cell tumors, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumors, hepatobiliary cancer, histiocytosis, anal cancer, melanoma, mature b-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, salivary gland cancer, renal cell carcinoma, and bone cancer.

[0066] In some embodiments, the cancer is a KRAS inhibitor-resistant cancer. In some embodiments, the KRAS inhibitor-resistant cancer comprises intrinsic resistance, acquired resistance or adaptive resistance. "KRAS-resistant cancer" can comprise cancer that has acquired resistance to one or more KRAS inhibitors due to previous treatment with one or more KRAS inhibitors, or cancer that has intrinsic resistance to one or more KRAS inhibitors.

[0067] In some embodiments, the acquired KRAS resistance is induced by a KRAS inhibitor.In some embodiments, the acquired KRAS resistance is induced by sotorasib (AMG510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and pharmaceutically acceptable salts thereof.In some embodiments, the KRAS inhibitor-resistant cancer is sotorasib (AMG510)-resistant cancer or adagrasib (MRTX849)-resistant cancer. Resistant cancers may be resistant from the start of treatment or may become resistant during treatment. In some embodiments, sotorasib resistance is induced by adagrasib.

[0068] In certain embodiments, the KRAS inhibitor resistant cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, small intestine cancer, colon adenocarcinoma, rectal adenocarcinoma, small intestine adenocarcinoma, lung adenocarcinoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, plasma cell myeloma, gallbladder cancer, anaplastic thyroid cancer, ampullary carcinoma, cervical cancer, gastrointestinal neuroendocrine tumors, uterine endometrioid carcinoma, germ cell tumors, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumor, hepatobiliary cancer, histiocytosis, anal cancer, melanoma, mature B-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, salivary gland cancer, renal cell carcinoma, and bone cancer.

[0069] As understood herein, the PIM kinase inhibitor and the KRAS inhibitor are each administered in an amount that is therapeutically effective in combination. In some embodiments, the dosage of the PIM kinase inhibitor and the KRAS inhibitor is more effective than either monotherapy treatment. In some embodiments, the dosage of the PIM kinase inhibitor and the KRAS inhibitor is synergistic against KRAS mutant cancer. In some embodiments, the KRAS mutant cancer has KRAS p.G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutation, or a combination thereof. In some embodiments, the cancer has KRAS p.G12C or G12D mutation, or a combination thereof.

[0070] In certain embodiments, the weight ratio of the PIM kinase inhibitor to the KRAS inhibitor is about 1:0.001 to 0.001:1, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, about 5:1 to about 1:5, or about 1:1 to about 1:5. In certain embodiments, the weight ratio of the PIM kinase inhibitor to the KRAS inhibitor is about 1:0.01, about 10:1, about 5:1, about 3:1, about 2:1, about 1:1, about 1:0.1, about 1:0.5, about 1:1, about 1:1.5, about 1:2, about 1:3, about 1:5, about 1:10, or about 1:100. In certain embodiments, the weight ratio of the PIM kinase inhibitor to the KRAS inhibitor is about 500:1, about 250:1, about 200:1, about 150:1, about 120:1, about 100:1, about 30:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 5:1, about 3:1, about 2:1, about 1:1, about 1:0.1, about 1:0.5, about 1:1, about 1:1.5, about 1:2, about 1:3, about 1:5, about 1:10, about 1:30, or about 1:100. Preferably, the KRAS inhibitor is a KRAS inhibitor. G12C inhibitor, KRAS G12V inhibitor, KRAS G12D inhibitor, KRAS G13C inhibitor, KRAS G13D inhibitor, KRAS Q61H inhibitor, KRAS Q61L inhibitor, KRAS Q61R inhibitor, KRAS K117Ninhibitor, pan-KRAS inhibitor, KRAS-SOS1 interaction inhibitor or KRAS signaling inhibitor.

[0071] In one embodiment, GDC-0570 and KRAS G12C The weight ratio of the inhibitors is about 1:0.001 to 0.001:1, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, about 5:1 to about 1:5, or about 1:1 to about 1:5. In one embodiment, the PIM kinase inhibitor and KRAS G12C The weight ratio of inhibitors is about 1:0.01, about 10:1, about 5:1, about 3:1, about 2:1, about 1.5:1, about 1:1, about 1:0.1, about 1:0.5, about 1:1, about 1:1.5, about 1:2, about 1:3, about 1:5, about 1:10, about 1:100. G12C The inhibitors are sotorasib or RG6330 (GDC-6036).

[0072] In one embodiment, GDC-0570 and KRAS G12D The weight ratio of the inhibitors is about 1:0.001 to 0.001:1, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, or about 1:1 to about 1:5. G12D The weight ratio of inhibitors is about 500:1, about 250:1, about 200:1, about 150:1, about 120:1, about 1:0.01, about 30:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 5:1, about 3:1, about 2:1, about 1:1, about 1:0.1, about 1:0.5, about 1:1, about 1:1.5, about 1:2, about 1:3, about 1:5, about 1:10, about 1:30, or about 1:100. G12D Inhibitors are MRTX1133 or KRAS G12D Cobimetinib is a MEK inhibitor, which inhibits KRAS G12D Inhibits signal transduction.

[0073] In one embodiment, GDC-0570 and KRAS G12VThe weight ratio of the inhibitors is about 1:0.001 to 0.001:1, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, or about 1:1 to about 1:5. G12V The weight ratio of inhibitors is about 500:1, about 250:1, about 200:1, about 150:1, about 120:1, about 1:0.01, about 30:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 5:1, about 3:1, about 2:1, about 1:1, about 1:0.1, about 1:0.5, about 1:1, about 1:1.5, about 1:2, about 1:3, about 1:5, about 1:10, or about 1:100. G12V Inhibitors are KRAS G12V Cobimetinib is a MEK inhibitor, which inhibits KRAS G12V Inhibits signal transduction.

[0074] In certain embodiments, the weight ratio of GDC-0570 to the KRAS signaling inhibitor is about 1:0.001 to 0.001:1, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, or about 1:1 to about 1:5. In some embodiments, the weight ratio of GDC-0570 to the KRAS signaling inhibitor is about 500:1, about 250:1, about 200:1, about 150:1, about 120:1, about 1:0.01, about 30:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 5:1, about 3:1, about 2:1, about 1:1, about 1:0.1, about 1:0.5, about 1:1, about 1:1.5, about 1:2, about 1:3, about 1:5, about 1:10, or about 1:100. In some embodiments, the KRAS signaling inhibitor is cobimetinib. Cobimetinib is a MEK inhibitor and is a KRAS G12D , KRAS G12V Inhibits all oncogenic KRAS downstream signaling, such as

[0075] In certain embodiments, the molar ratio of the PIM kinase inhibitor to the KRAS inhibitor is about 1:1000, about 1:100, about 1:10, or about 1:5. In certain embodiments, the molar ratio of the PIM kinase inhibitor to the KRAS inhibitor is about 1:0.001 to about 1:1000, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, or about 1:1 to about 1:5.

[0076] In some embodiments, the PIM kinase inhibitor and the KRAS inhibitor are administered simultaneously. In some embodiments, the PIM kinase inhibitor and the KRAS inhibitor are administered sequentially. When administered sequentially, the combination can be administered in two or more doses. Combined administration includes simultaneous administration using separate preparations and sequential administration in any order, where preferably there is a period when both (or all) active agents simultaneously exert their biological activity.

[0077] Suitable dosages of any of the above co-administered agents are those currently in use and may be reduced by the combined action (synergy) of newly identified agents with other chemotherapeutic agents or treatments, such as to increase the therapeutic index or reduce toxicity or other side effects or consequences.

[0078] In further embodiments, the method may further comprise surgical treatment and / or radiation therapy. The amounts and relative timing of administration of the PIM kinase inhibitor, KRAS inhibitor and other pharmaceutically active chemotherapeutic agent are selected to achieve the desired combined therapeutic effect.

[0079] In certain embodiments, the present invention provides a method of treating a cancer harboring a KRAS mutation in a human subject in need thereof, comprising administering a therapeutically effective amount of N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide (GDC-0570) or a pharmaceutically acceptable salt thereof. In certain embodiments, the present invention provides GDC-0570 or a pharmaceutically acceptable salt thereof for use as a medicament for treating a cancer harboring a KRAS mutation. In certain embodiments, the present invention provides the use of GDC-0570 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a cancer harboring a KRAS mutation. In certain embodiments, GDC-0570 can be used as a single active agent for various KRAS mutant cancers. In some embodiments, the KRAS mutation is selected from G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutations. In some embodiments, the cancer is resistant to a KRAS inhibitor, including intrinsic, acquired, or adaptive resistance. In some embodiments, the cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, small intestine cancer, colon adenocarcinoma, rectal adenocarcinoma, small intestine adenocarcinoma, lung adenocarcinoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, plasma cell myeloma, gallbladder cancer, anaplastic thyroid cancer, ampullary carcinoma, cervical cancer, gastrointestinal neuroendocrine tumors, uterine endometrioid carcinoma, germ cell tumors, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumors, hepatobiliary cancer, histiocytosis, anal cancer, melanoma, mature B-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, salivary gland cancer, renal cell carcinoma, and bone cancer.In certain embodiments, the KRAS inhibitor resistant cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, small intestine cancer, colon adenocarcinoma, rectal adenocarcinoma, small intestine adenocarcinoma, lung adenocarcinoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, plasma cell myeloma, gallbladder cancer, anaplastic thyroid cancer, ampullary carcinoma, cervical cancer, gastrointestinal neuroendocrine tumors, uterine endometrioid carcinoma, germ cell tumors, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumor, hepatobiliary cancer, histiocytosis, anal cancer, melanoma, mature B-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, salivary gland cancer, renal cell carcinoma, and bone cancer.

[0080] 3. Combinations, Pharmaceutical Compositions and Kits In another embodiment, the present invention relates to a combination comprising a PIM kinase inhibitor and a KRAS inhibitor, preferably for use in the treatment of cancer, such as a KRAS mutant cancer or a KRAS inhibitor resistant cancer.

[0081] In some embodiments, the combination is provided in a single pharmaceutical composition together with pharmaceutically acceptable additives.In other embodiments, the combination is provided in two pharmaceutical compositions that are administered together, one of which comprises a PIM kinase inhibitor and a pharmaceutically acceptable additive, and the other of which comprises a KRAS inhibitor and a pharmaceutically acceptable additive.

[0082] As used herein, a pharmaceutically acceptable excipient refers to a substance that aids in the in vivo delivery and / or manufacture of a pharmaceutical composition containing one or more active agents described herein. Pharmaceutically acceptable excipients are inert. Non-limiting examples of pharmaceutically acceptable excipients include pharmaceutically acceptable polymers, water, NaCl, saline solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, surfactants, coatings, sweeteners, flavoring agents, salt solutions, alcohol, oils, gelatin, carbohydrates, coloring agents, etc. Such formulations may be sterilized and, if desired, mixed with auxiliary substances such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring and / or fragrance substances. Pharmaceutically acceptable excipients are described in Handbook of Pharmaceutical Excipients, 8 th Edition, published by the Pharmaceutical Press (2017) and the U.S. Food and Drug Administration Inactive Ingredients Database (July 2017), which are incorporated herein by reference.

[0083] Pharmaceutical compositions may be packaged in a variety of ways depending on the method used to administer the drug. Generally, the distribution item includes a container having the pharmaceutical formulation in an appropriate form deposited therein. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, ampoules, plastic bags, metal cylinders, and the like. The container may also include a tamper-evident structure to prevent inadvertent use of the contents of the package. Additionally, the container is provided with a label describing the contents of the container. The label may also include appropriate warnings.

[0084] Pharmaceutical compositions can be prepared for various routes and types of administration. Pharmaceutical compositions are taken and administered in a manner consistent with good clinical practice, i.e., dosage, concentration, schedule, course, vehicle and route of administration. Factors to consider in this context include the specific disorder to be treated, the specific mammal to be treated, the clinical condition of each patient, the cause of the disorder, the drug delivery site, administration method, administration schedule and other factors known to medical professionals.

[0085] In some embodiments, pharmaceutical compositions are formulated for oral delivery.The formulation of PIM kinase inhibitor and / or KRAS inhibitor suitable for oral administration can be prepared as separate units such as pills, hard or soft, for example, gelatin capsules, cachets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, syrups or elixirs, each of which contains a predetermined amount of PIM kinase inhibitor and / or KRAS inhibitor.The amount of the compound of PIM kinase inhibitor and KRAS inhibitor can be formulated into pills, capsules, solution or suspension as combined preparation.Alternatively, PIM kinase inhibitor and KRAS inhibitor can be separately formulated into pills, capsules, solution or suspension for alternate administration.

[0086] In an embodiment, the pharmaceutical composition is an orally administered solid dosage form, such as a tablet, capsule, or pill. In an embodiment, the solid dosage form is a tablet.

[0087] The dose can be administered once a day (QD), twice a day (BID), or more frequently, depending on the pharmacokinetic (PK) and pharmacodynamic (PD) properties, including absorption, distribution, metabolism, and excretion, of a particular compound.In addition, toxicity factors can affect the dosage and administration regimen.When administered orally, pills, capsules, or tablets can be taken twice a day, daily, or less frequently, such as weekly, or once every two or three weeks, for a certain period of time.The regimen can be repeated for several treatment cycles.

[0088] In another aspect, the present invention provides a kit comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable excipient.

[0089] The kit may further include a label or package insert on or in association with the container. The term "package insert" refers to instructions customarily included in commercial packaging of a therapeutic product, including information about the indications, use, dosage, administration, contraindications, and / or warnings associated with the use of such a therapeutic product. Suitable containers include, for example, bottles, vials, syringes, blister packs, and the like. The container may be formed from a variety of materials, such as glass or plastic. The container may hold a compound or a pharmaceutically acceptable salt thereof or a formulation thereof that is effective for treating a condition and may have a sterile access port (e.g., the container may be an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a PIM kinase inhibitor and / or a PIM kinase inhibitor compound. The label or package insert indicates that the composition is used to treat a selected condition. Alternatively or additionally, the product may further include a second container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0090] The kit may further comprise a PIM kinase inhibitor and instructions for administration of the PIM kinase inhibitor. For example, if the kit comprises a PIM kinase inhibitor and a first composition comprising a PIM kinase inhibitor, the kit may further comprise instructions for simultaneous, sequential, or separate administration of the PIM kinase inhibitor and the KRAS inhibitor to a subject in need thereof.

[0091] In other cases, the kit is suitable for delivering a solid oral form of the PIM kinase inhibitor and / or PIM kinase inhibitor, such as tablets or capsules. Such kits preferably contain several unit doses. Such kits may include a card with the dosages oriented in the order of intended use. An example of such a kit is a "blister pack." Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms. Optionally, memory aids, such as numbers, letters, or other markings, or a calendar designating the days in the treatment schedule when the dosages can be administered, may be inserted.

[0092] The kit may include (a) a first container containing a PIM kinase inhibitor; and (b) a second container containing a KRAS inhibitor. Alternatively or additionally, the kit may further include a third container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0093] When the kit includes a PIM kinase inhibitor and a PIM kinase inhibitor composition, the kit can include containers containing the separate compositions, such as individual bottles or individual foil packets; however, the separate compositions may also be contained in a single, undivided container. Typically, the kit includes instructions for administering the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), administered at different dosage intervals, or when titration of the individual components of the combination is desired by the treating physician. [Example]

[0094] The present invention will be further described and illustrated with reference to the following examples. It should be noted that the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

[0095] The various PDX models of the present invention were established at GenenDesign Co., Ltd. (Shanghai, China). The various PDX models used in the examples are summarized in Table 1. [Table 1]

[0096] Example 1. Sotorasib-sensitive KRAS G12C Combination Study of GDC-0570 and Sotorasib in the Mutant Colorectal Cancer Patient-Derived Xenograft (PDX) Model CRC024 This trial is G12C This study aimed to evaluate the in vivo combined antitumor efficacy of GDC-0570 and its combination with sotorasib in the mutant colorectal cancer (CRC) patient-derived xenograft (PDX) model CRC024. The CRC024 PDX model was a sotorasib-sensitive model derived from a 75-year-old Chinese female CRC cancer patient. KRAS in CRC PDX Models G12C Mutations were confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were divided into four groups: vehicle, 300 mg / kg GDC-0570, 100 mg / kg sotorasib, and 300 mg / kg GDC-0570 + 100 mg / kg sotorasib. The dosing solution was administered orally daily. The study period was 27 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0097] GDC-0570 medium: 0.5% methylcellulose (MC) and 0.2% Tween-80 (MCT), maintained at 2–8°C.

[0098] GDC-0570 Dosing Solution: Weigh the required amount of GDC-0570 powder and mix it with the appropriate amount of 0.5% MC 0.2% Tween 80 solution in a container to achieve a GDC-0570 concentration of 60 mg / ml. Vortex and sonicate until homogenous. Store the dosing suspension at 2-8°C for up to 1 week.

[0099] Sotorasib vehicle: 50% w / w polyethylene glycol 400 (PEG400) + 50% w / w propylene glycol (PG).

[0100] Sotorasib Dosing Solution: The required amount of sotorasib powder was weighed into a container. A 50% / 50% PEG400 / PG mixture was then added to the container to achieve a sotorasib concentration of 20 mg / ml. 1 N hydrochloric acid solution was added to the same container to a final concentration of 0.39%. The sotorasib was dispersed by vortexing for 5 minutes, followed by sonication for 10 minutes until completely dissolved. The dosing solution was stored at 2-8°C for up to 1 week.

[0101] Measurements and Calculations Tumor size was measured twice a week, while body weight was measured daily before administration. Clinical signs were observed daily. All animals were sacrificed on day 27 after tumor size calibration. After the animals were sacrificed, tumor samples were collected. Tumor volume (TV) = (length x width) 2 ) / 2 Relative tumor volume (RTV) = TV f / TV0 (wherein TV0 and TV f are the tumor volumes measured on days 0 and 27, respectively); Tumor growth inhibition T / C ratio (%) = (RTV of treatment group / RTV of vehicle control group) × 100%; Tumor growth inhibition rate (TGI) TGI = [1-(TVt f -TVt0) / (TVc f -TVc0)] × 100% TVt f was the group mean tumor volume (TV) of the treatment group on the last treatment day TVt0 was the group mean TV for the treatment group on treatment day 0 TVc f The group mean TV for the control group on the final treatment day was TVc0 was the group mean TV of the control group on day 0 of treatment. Percent tumor regression (% regression) = 100 × (TV0 - TV f ) / TV0 TV0 was the group mean TV measured for the same group but on treatment day 0 TV f was the group mean TV measured in the same group but on the last treatment day Percentage body weight change (%BWC) = (BWc - BWi) / BWi x 100% (where "c" means current, "i" means initial, and "BW" means body weight).

[0102] Data analysis Tumor growth curves were plotted using tumor volume as the Y-axis and time as the X-axis; body weight change curves were plotted using animal weight as the Y-axis and time as the X-axis. Data on tumor volume and percentage body weight change were analyzed using one-way ANOVA followed by a significant difference test using Bartlett's test (p<0.05).

[0103] Criteria used to evaluate the tumor growth inhibitory effect of test products based on tumor growth inhibition TGI rate: Significant tumor inhibition effect: TGI(%)>60% and P<0.05; Insignificant tumor inhibitory effect: TGI (%)≦60% or P>0.05

[0104] Test results Tables 2, 3, 4 and Figure 1 show KRAS G12C Shows the effect of GDC-0570 and sotorasib on tumor growth in the mutant CRC PDX model CRC024.

[0105] KRAS G12C Under the current study conditions in the mutant CRC PDX model, the 300 mg / kg GDC-0570, 100 mg / kg sotorasib monotherapy treatment groups, and the 300 mg / kg GDC-0570 + 100 mg / kg sotorasib combination group all demonstrated significant tumor inhibition effects. Furthermore, the GDC-0570 and sotorasib combination group demonstrated superior efficacy compared to either monotherapy treatment. Furthermore, the GDC-0570 and sotorasib combination group demonstrated superior efficacy compared to either monotherapy treatment.

[0106] No mice in any dose group experienced severe weight loss (defined as a loss of more than 20% of body weight) or other abnormalities throughout the study.

[0107] [Table 2] " / " means not applicable

[0108] [Table 3] " / " means not applicable; P<0.05 indicates statistical significance.

[0109] [Table 4] P<0.05 indicates statistical significance.

[0110] Example 2. Sotorasib-insensitive KRAS G12C Combination study of GDC-0570 and sotorasib in the mutant NSCLC cancer patient-derived xenograft (PDX) model LUN055 This trial is G12C This study was conducted to evaluate the in vivo combined antitumor efficacy of GDC-0570 and its combination with sotorasib in the LUN055 patient-derived xenograft (PDX) model of NSCLC. The LUN055 PDX model was derived from a 60-year-old Chinese male NSCLC cancer patient and was a sotorasib-insensitive model. KRAS in NSCLC PDX Models G12C Mutations were confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were divided into four groups: vehicle, 300 mg / kg GDC-0570, 100 mg / kg sotorasib, and 300 mg / kg GDC-0570 + 100 mg / kg sotorasib. The dosing solution was administered orally daily. The study period was 28 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0111] The GDC-0570 vehicle, GDC-0570 dosing solution, sotorasib vehicle, sotorasib dosing solution, measurements and calculations, and data analysis were the same as in Example 1.

[0112] Test results Tables 5 to 7 and Figure 2 show KRAS G12C Shows the effect of GDC-0570 and sotorasib on tumor growth in mutant NSCLC PDX model LUN055.

[0113] KRAS G12C Under the current study conditions in the mutant NSCLC PDX model, monotherapy treatment with 300 mg / kg GDC-0570 and 100 mg / kg sotorasib did not demonstrate significant tumor growth inhibition. In contrast, the combination of 300 mg / kg GDC-0570 and 100 mg / kg sotorasib demonstrated significant tumor inhibition. Furthermore, the GDC-0570 and sotorasib combination demonstrated superior efficacy compared to monotherapy treatment.

[0114] No mice in any dose group experienced severe weight loss (defined as a loss of more than 20% of body weight) or other abnormalities throughout the study.

[0115] [Table 5] " / " means not applicable

[0116] [Table 6] " / " means not applicable; P<0.05 indicates statistical significance.

[0117] [Table 7] P<0.05 indicates statistical significance.

[0118] Example 3. GDC-0570 and sotorasib combination inhibits sotorasib-sensitive KRAS G12C Prevented tumor regrowth after treatment ended on day 28 in the mutant NSCLC cancer patient-derived xenograft (PDX) model LUN156 This study was conducted to evaluate the in vivo antitumor efficacy of GDC-0570 and sotorasib in the NSCLC PDX model LUN156. The LUN156 PDX model was a sotorasib-sensitive model derived from a 73-year-old Chinese male NSCLC cancer patient. KRAS in LUN156 G12C Mutations were confirmed by whole-exome sequencing and PCR sequencing.

[0119] Tumor-bearing mice were divided into four groups: vehicle, 300 mg / kg GDC-0570, 100 mg / kg sotorasib, and 300 mg / kg GDC-0570 + 100 mg / kg sotorasib. The dosing solution was orally administered daily until day 28. Dosing was stopped after day 28, and tumor growth observation continued until day 45. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0120] GDC-0570 vehicle, GDC-0570 dosing solution, sotorasib vehicle, sotorasib dosing solution, measurements and calculations, and data analysis were similar to Example 1.

[0121] Test results Tables 8 to 11 and Figure 3 show the results of sotorasib-sensitive KRAS G12C Shows the effect of GDC-0570 and sotorasib on tumor growth in mutant NSCLC PDX models.

[0122] Compared with the vehicle group, the 300 mg / kg GDC-0570, 100 mg / kg sotorasib monotherapy treatment groups, and their combination treatments all demonstrated significant tumor growth inhibitory effects (Tables 8-11). The GDC-0570 and sotorasib combination group demonstrated superior efficacy compared with the GDC-0570 monotherapy treatment, which was statistically significant (Table 10). Although the TGI effect between the combination group and the sotorasib monotherapy group was not statistically significant based on the p-value during the 28-day dosing period (Table 10), only one tumor showed complete regression in the sotorasib monotherapy group, whereas all (5 / 5) tumors in the combination group completely regressed by day 28. Furthermore, after dosing was stopped on day 28, all (5 / 5) tumors in both the GDC-0570 and sotorasib monotherapy groups experienced regrowth, whereas only one small tumor reappeared in the combination group, which remained at a minimal volume until day 45. Therefore, the combination of GDC-0570 and sotorasib prevented tumor regrowth after the discontinuation of sotorasib monotherapy.In conclusion, the GDC-0570 and sotorasib combination group demonstrated superior efficacy compared to monotherapy treatment.

[0123] No mice in any dose group experienced severe weight loss (defined as a loss of more than 20% of body weight) or other abnormalities throughout the study.

[0124] [Table 8] " / " means not applicable

[0125] [Table 9] " / " means not applicable; P<0.05 indicates statistical significance.

[0126] [Table 10] P<0.05 indicates statistical significance.

[0127] [Table 11] "CR" means complete regression

[0128] Example 4. GDC-0570 and sotorasib combination study in the KRASG12C NSCLC model with acquired resistance to sotorasib LUN2156-44 This trial is G12C This study was conducted to evaluate the in vivo combined antitumor efficacy of GDC-0570 and its combination with sotorasib in the mutant NSCLC patient-derived xenograft (PDX) model LUN2156-44. The LUN2156-44 PDX model was a sotorasib-resistant model derived from the LUN156 PDX model. KRAS in LUN2156-44 G12C Mutations were confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were divided into six groups: vehicle, 150 mg / kg GDC-0570, 100 mg / kg sotorasib, 50 mg / kg GDC-0570 + 100 mg / kg sotorasib, 100 mg / kg GDC-0570 + 100 mg / kg sotorasib, and 150 mg / kg GDC-0570 + 100 mg / kg sotorasib. The dosing solution was administered orally daily. The study period was 28 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0129] GDC-0570 vehicle, GDC-0570 dosing solution, sotorasib vehicle, sotorasib dosing solution, measurements and calculations, data analysis were similar to Example 1.

[0130] Test results Tables 12 to 14 and Figure 4 show KRAS G12C Shows the effect of GDC-0570 and sotorasib on tumor growth in mutant NSCLC PDX model LUN2156-44.

[0131] At day 28, compared with the sotorasib-sensitive PDX model LUN156, the sotorasib-resistant PDX model LUN2156-44 reduced sotorasib single-agent activity from complete regression to 88% TGI and reduced GDC-0570 single-agent activity from plateau to <50% TGI.

[0132] Tumor volume measurements on day 28 showed that 150 mg / kg GDC-0570 treatment resulted in 38% tumor growth inhibition (TGI) and a 71% T / C ratio. 100 mg / kg sotorasib treatment resulted in 88% TGI and a 33% T / C ratio. The combination of 50 mg / kg GDC-0570 and 100 mg / kg sotorasib resulted in 92% tumor regression and a 16% T / C ratio. The combination of 100 mg / kg GDC-0570 and 100 mg / kg sotorasib resulted in 95% tumor regression and a 1% T / C ratio. The combination of 150 mg / kg GDC-0570 and 100 mg / kg sotorasib resulted in 98% tumor regression and a nearly 0% T / C ratio. Compared with the vehicle group, 150 mg / kg GDC-0570 monotherapy treatment did not show significant tumor growth inhibitory effects. The 100 mg / kg sotorasib monotherapy treatment group and its combination with three different doses of GDC-0570 demonstrated significant tumor-inhibitory effects. Furthermore, the combination of 100 mg / kg sotorasib with 50 mg / kg, 100 mg / kg, or 150 mg / kg GDC-0570 demonstrated superior efficacy compared with 100 mg / kg sotorasib monotherapy treatment. Complete regression was observed at all combination doses tested, even with 50 mg / kg GDC-0570. The combination reduced the GDC-0570 dose required for complete regression and produced a strong synergistic effect.

[0133] No mice in any dose group experienced severe weight loss (defined as a loss of more than 20% of body weight) or other abnormalities throughout the study.

[0134] [Table 12] " / " means not applicable

[0135] [Table 13] " / " means not applicable P<0.05 indicates statistical significance.

[0136] [Table 14] P<0.05 indicates statistical significance.

[0137] Example 5. KRAS G12C Combination Study of GDC-0570 and GDC-6036 in the Mutant Colorectal Cancer Patient-Derived Xenograft (PDX) Model CRC022 This trial is G12C This study was conducted to evaluate the in vivo combined antitumor efficacy of GDC-0570 and its combination with GDC-6036 in the mutant CRC patient-derived xenograft (PDX) model CRC022. This PDX model was derived from a 49-year-old Chinese female CRC cancer patient. KRAS in CRC#022 G12C Mutations were confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were divided into four groups: vehicle, 100 mg / kg QD GDC-6036, 300 mg / kg QD GDC-0570, and 300 mg / kg GDC-0570 + 100 mg / kg GDC-6036. The study period was 27 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0138] GDC-0570 vehicle, GDC-0570 dosing solution, measurements and calculations, data analysis were similar to Example 1.

[0139] GDC-6036 Media: 0.5% Methocel

[0140] GDC-6036 Dosing Solution: The appropriate amount of GDC-6036 was weighed into a container. The appropriate volume of 0.5% Methocel was added to the container to achieve 20 mg / ml GDC-6036. A homogenous solution was obtained by repeated vortexing and sonication (>1 hour). The dosing solution was stored at 2-8°C for up to 1 week.

[0141] Test results Tables 15-16 and Figure 5 show KRAS G12C Shows the effect of GDC-0570 and GDC-6036 on tumor growth in mutant colorectal PDX models.

[0142] Tumor volume measurements on day 27 showed that the 100 mg / kg QD GDC-6036 group had 78% tumor growth inhibition (TGI) and a 30% T / C ratio. The 300 mg / kg QD GDC-0570 group had 79% tumor growth inhibition (TGI) and a 29% T / C ratio. The combination of 300 mg / kg GDC-0570 + 100 mg / kg GDC-6036 had 99% tumor growth inhibition (TGI) and a 11% T / C ratio.

[0143] Compared with the vehicle group, all monotherapy and combination groups demonstrated significant tumor inhibition effects. The GDC-0570 monotherapy group demonstrated moderate to strong tumor inhibition effects (70-80% TGI). The GDC-0570 and GDC-6036 combination treatment demonstrated superior efficacy compared with GDC-0570 or GDC-6036 monotherapy treatment.

[0144] No mice in any dose group experienced severe weight loss (defined as a loss of more than 20% of body weight) or other abnormalities throughout the study. The combination of GDC-0570 and GDC-6036 was well tolerated in mice (<5% weight loss).

[0145] [Table 15] " / " means not applicable

[0146] [Table 16] " / " means not applicable; P<0.05 indicates statistical significance.

[0147] Example 6. KRAS G12C Dose-dependent single-agent activity study of GDC-0570 in the mutant NSCLC PDX model LUN156 This study was conducted to evaluate the in vivo antitumor efficacy of GDC-0570 in the NSCLC patient-derived xenograft (PDX) model LUN156. Tumor-bearing mice were divided into five dose groups, including vehicle, 50 mg / kg GDC-0570, 100 mg / kg GDC-0570, 200 mg / kg GDC-0570, and 300 mg / kg GDC-0570. The dosing solution was orally administered daily. The study period was 21 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0148] GDC-0570 media, measurements and calculations, data analysis were similar to Example 1.

[0149] GDC-0570 Dosing Solution: The 60 mg / ml GDC-0570 dosing solution from Example 1 was further diluted with MCT to other concentrations (10 mg / ml, 20 mg / ml, and 40 mg / ml). The dosing suspension was stored at 2-8°C for up to 1 week.

[0150] Test results Tables 17-18 and Figure 6 show KRAS G12C Shows the effect of GDC-0570 on tumor growth in the mutant NSCLC PDX model LUN156.

[0151] Under the current study conditions in the LUN156 model, all GDC-0570 dose levels of 50, 100, 200, and 300 mg / kg demonstrated significant inhibition of tumor growth (TGI >60% and p-value <0.05). Mice in all dose groups did not experience severe weight loss (defined as a loss of more than 20% of body weight) or other abnormalities throughout the study, indicating that all doses of GDC-0570 were well tolerated.

[0152] [Table 17] " / " means not applicable

[0153] [Table 18] " / " means not applicable; P<0.05 indicates statistical significance.

[0154] Example 7. KRAS G12 Dose-dependent single-agent activity study of GDC-0570 in the D-mutant NSCLC PDX model LUN#137 This study was conducted to evaluate the in vivo antitumor efficacy of GDC-0570 in the NSCLC patient-derived xenograft (PDX) model LUN#137. This PDX model was derived from a 57-year-old Chinese male NSCLC patient. KRAS in LUN#137 G12 The D mutation was confirmed by whole-exome sequencing and PCR sequencing.

[0155] Tumor-bearing mice were divided into five dose groups, including a vehicle group, 50 mg / kg GDC-0570, 100 mg / kg GDC-0570, 200 mg / kg GDC-0570, and 300 mg / kg GDC-0570 groups. Dosing solutions were orally administered daily. The study period was 22 days. Tumor volumes were measured twice weekly. Body weights were measured each day before dosing.

[0156] GDC-0570 vehicle, measurements and calculations, data analysis are similar to Example 1. GDC-0570 dosing solution is similar to Example 6.

[0157] Test results Tables 19-20 and Figure 7 show KRAS G12 Shows the effect of GDC-0570 on tumor growth in D mutant NSCLC PDX model LUN#137.

[0158] Under the current study conditions in the LUN#137 model, GDC-0570 demonstrated dose-dependent single-agent antitumor activity, with significant tumor-inhibitory effects observed at the 200 mg / kg and 300 mg / kg dose levels (TGI >60% and p-value <0.05). Mice in all dose groups did not experience severe weight loss (defined as a loss of >20% of body weight) or other abnormalities throughout the study, indicating that all doses of GDC-0570 were well tolerated.

[0159] [Table 19] " / " means not applicable

[0160] [Table 20] " / " means not applicable; P<0.05 indicates statistical significance.

[0161] Example 8. KRAS G12D GDC-0570 and Cobimetinib Combination Study in the Pancreatic Cancer Model PAN092 This trial is G12 This study aimed to evaluate the in vivo combined antitumor efficacy of GDC-0570 and its combination with the MEK inhibitor cobimetinib in the KRAS D-mutant pancreatic patient-derived xenograft (PDX) model PAN092. This PDX model was derived from a 65-year-old Chinese male pancreatic cancer patient. G12The D mutation was confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were randomized into treatment groups (5 mice per group) containing vehicle, 2.5 mg / kg QD cobimetinib, 300 mg / kg QD GDC-0570, and 300 mg / kg GDC-0570 + 2.5 mg / kg cobimetinib. The study duration was 27 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0162] GDC-0570 vehicle, GDC-0570 dosing solution, measurements and calculations, data analysis were similar to Example 1.

[0163] Cobimetinib vehicle: 0.5% methylcellulose / 0.2% Tween-80 (MCT)

[0164] Cobimetinib Dosing Solution: The appropriate amount of cobimetinib was weighed into a container. The appropriate volume of MCT was added to the container to achieve 0.5 mg / ml cobimetinib. A homogenous solution was obtained by repeated vortexing and sonication. The dosing solution was stored at 2-8°C for up to 1 week.

[0165] Test results Tables 21 to 23 and Figure 8 show KRAS G12 Shows the effect of GDC-0570 and cobimetinib on tumor growth in the D mutant pancreatic PDX model PAN092.

[0166] Tumor volume measurements on day 27 showed that the 2.5 mg / kg QD cobimetinib group had 65% tumor growth inhibition (TGI) and a 47% T / C ratio. The 300 mg / kg QD GDC-0570 group had 55% tumor growth inhibition (TGI) and a 56% T / C ratio. The combination of 300 mg / kg GDC-0570 + 2.5 mg / kg cobimetinib had >100% tumor growth inhibition (TGI) and a 19% T / C ratio.

[0167] Compared to the vehicle group, all monotherapy and combination groups demonstrated significant tumor inhibition. The cobimetinib monotherapy group demonstrated a moderate tumor inhibition effect (65% TGI). The GDC-0570 monotherapy group also demonstrated a moderate tumor inhibition effect (55% TGI). GDC-0570 and cobimetinib combination treatment demonstrated significantly superior efficacy compared to GDC-0570 or cobimetinib monotherapy treatment.

[0168] [Table 21] " / " means not applicable

[0169] [Table 22] " / " means not applicable; P<0.05 means statistical significance.

[0170] [Table 23] P<0.05 indicates statistical significance.

[0171] Example 9. KRAS G12D GDC-0570 and MRTX1133 Combination Study in the Pancreatic Cancer Model PAN092 This trial is G12 GDC-0570 and KRAS in the D-mutant pancreatic patient-derived xenograft (PDX) model PAN092 G12 This study was conducted to evaluate the in vivo antitumor efficacy of the KRAS inhibitor MRTX1133 in combination with PAN092. This PDX model was derived from a 65-year-old Chinese male pancreatic cancer patient. G12The D mutation was confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were randomized into treatment groups (5 mice per group) containing vehicle, 10 mg / kg QD MRTX1133, 300 mg / kg QD GDC-0570, and 300 mg / kg GDC-0570 + 10 mg / kg MRTX1133. The study period was 27 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0172] GDC-0570 vehicle, GDC-0570 dosing solution, measurements and calculations, data analysis were similar to Example 1.

[0173] MRTX1133 medium: 10% Captisol in 50 mM citrate buffer, pH 5.0

[0174] MRTX1133 Dosing Solution: The appropriate amount of MRTX1133 was weighed into a container. The appropriate volume of vehicle was added to the container to achieve a 2 mg / ml MRTX1133 formulation. A homogenous solution was obtained by repeated vortexing and sonication (>1 hour). The dosing solution was stored at 2-8°C for up to 1 week.

[0175] Test results Tables 24-25 and Figure 9 show KRAS G12 Shows the effect of GDC-0570 and MRTX1133 on tumor growth in D mutant pancreatic PDX models.

[0176] Tumor volume measurements on day 27 showed that the 10 mg / kg QD MRTX1133 group had 74% tumor growth inhibition (TGI) and a 40% T / C ratio (%). The 300 mg / kg QD GDC-0570 group had 55% tumor growth inhibition (TGI) and a 56% T / C ratio (%). The combination of 300 mg / kg GDC-0570 + 10 mg / kg MRTX1133 had 89% tumor growth inhibition (TGI) and a 27% T / C ratio (%).

[0177] Compared to the vehicle group, all monotherapy and combination groups demonstrated significant tumor inhibition effects. GDC-0570 and MRTX1133 monotherapy groups demonstrated weak to moderate tumor inhibition effects (74% and 55% TGI, respectively). GDC-0570 and MRTX1133 combination treatment demonstrated superior efficacy compared to GDC-0570 monotherapy treatment.

[0178] [Table 24] " / " means not applicable

[0179] [Table 25] " / " means not applicable; P<0.05 means statistical significance.

[0180] Example 10. KRAS G12 GDC-0570 and cobimetinib combination study in V-mutant colorectal cancer PDX model CRC051 This trial is G12 This study was conducted to evaluate the in vivo combined antitumor efficacy of GDC-0570 and its combination with cobimetinib in the KRAS V mutant colorectal patient-derived xenograft (PDX) model CRC051. This PDX model was derived from a Chinese colorectal cancer patient. G12 The V mutation was confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were divided into four groups: vehicle, 2.5 mg / kg QD cobimetinib, 300 mg / kg QD GDC-0570, and 300 mg / kg GDC-0570 + 2.5 mg / kg cobimetinib. The study period was 27 days. Tumor volume was measured twice weekly. Body weight was measured each day before dosing.

[0181] The GDC-0570 vehicle, GDC-0570 dosing solution, measurements and calculations, and data analysis were the same as in Example 1.

[0182] The cobimetinib vehicle and cobimetinib dosing solution are the same as in Example 8.

[0183] Test results Tables 26-28 and Figure 10 show KRAS G12 Shows the effect of GDC-0570 and cobimetinib on tumor growth in a V mutant colorectal PDX model.

[0184] Tumor volume measurements on day 27 showed that the 2.5 mg / kg QD cobimetinib group had 4% tumor growth inhibition (TGI) and a 97% T / C ratio. The 300 mg / kg QD GDC-0570 group had 52% tumor growth inhibition (TGI) and a 63% T / C ratio. The combination of 300 mg / kg GDC-0570 + 2.5 mg / kg cobimetinib had >100% tumor growth inhibition (TGI) and a 26% T / C ratio.

[0185] Compared with the vehicle group, only the combination group showed a significant tumor growth inhibitory effect, while none of the monotherapy groups showed a significant tumor inhibitory effect.

[0186] GDC-0570 and cobimetinib combination treatment demonstrated superior efficacy compared with GDC-0570 or cobimetinib monotherapy treatment.

[0187] [Table 26] " / " means not applicable

[0188] [Table 27]

[0189] [Table 28] P<0.05 indicates statistical significance.

[0190] While the foregoing sets forth certain details by way of explanation and example, for purposes of clarity of understanding, the specification and examples should not be construed as limiting the scope of the invention. All patent and scientific literature cited herein is hereby incorporated by reference in its entirety.

Claims

1. 1. A method of treating cancer in a human subject in need thereof, comprising administering to the subject therapeutically effective amounts of a PIM kinase inhibitor and a KRAS inhibitor.

2. 2. The method of claim 1, wherein the PIM kinase inhibitor is a PIM-1 kinase inhibitor, a PIM-2 kinase inhibitor, or a PIM-3 inhibitor.

3. The method of claim 2 , wherein the PIM kinase inhibitor is a pan-PIM kinase inhibitor.

4. 3. The method of claim 2, wherein the PIM kinase inhibitor is selected from the group consisting of AZD1208, LGH447, GDC-0570, GNE-1571, GNE-5775, GDC-0339, GNE-5652 and pharmaceutically acceptable salts thereof.

5. 4. The method of claim 3, wherein the PIM kinase inhibitor is selected from the group consisting of GDC-0570, GNE-1571, GNE-5775, GDC-0339, GNE-5652, and pharmaceutically acceptable salts thereof.

6. KRAS inhibitors inhibit KRAS Ｇ１２Ｃ Inhibitor, KRAS Ｇ１２Ｖ Inhibitor, KRAS Ｇ１２Ｄ Inhibitor, KRAS Ｇ１３Ｃ Inhibitor, KRAS Ｇ１３Ｄ Inhibitor, KRAS Ｑ６１Ｈ Inhibitor, KRAS Ｑ６１Ｌ Inhibitor, KRAS Ｑ６１Ｒ Inhibitor, KRAS Ｋ１１７Ｎ The method of any one of claims 1 to 5, wherein the inhibitor is selected from the group consisting of a KRAS inhibitor, a pan-KRAS inhibitor, a KRAS-SOS1 interaction inhibitor, or a KRAS signaling inhibitor.

7. KRAS Ｇ１２Ｃ the inhibitor is selected from the group consisting of sotorasib (AMG510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and pharmaceutically acceptable salts thereof; Ｇ１２Ｄ inhibitors selected from the group consisting of MRTX1133, JAB-22000, RMC-9805 (RM-036) and pharmaceutically acceptable salts thereof; Ｇ１３Ｃ The inhibitor is RMC-8839 and its pharmaceutically acceptable salts; KRAS Ｇ１２Ｖ The method of claim 6, wherein the inhibitor is selected from JAB-23000 and a pharmaceutically acceptable salt thereof; the pan-KRAS inhibitor is selected from the group consisting of RMC-6236, BBP-454, and a pharmaceutically acceptable salt thereof; the KRAS-SOS1 interaction inhibitor is selected from the group consisting of BI1701963, BAY-293, RMC-5845, BI-3406, SDGR5, and a pharmaceutically acceptable salt thereof; and the KRAS signaling inhibitor is selected from cobimetinib and a pharmaceutically acceptable salt thereof.

8. the PIM kinase inhibitor is selected from the group consisting of GDC-0570, GNE-1571, GNE-5775, GDC-0339, and GNE-5652, and pharmaceutically acceptable salts thereof; and Ｇ１２Ｃ 7. The method of claim 6, wherein the inhibitor is selected from the group consisting of sotorasib (AMG510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and pharmaceutically acceptable salts thereof.

9. The method of any one of claims 1 to 8, wherein the weight ratio of the PIM kinase inhibitor to the KRAS inhibitor is about 1:0.001 to 0.001:1, about 1:0.01 to about 1:100, about 1:0.1 to about 1:10, or about 1:5 to about 1:

5.

10. 10. The method of any of claims 1 to 9, wherein the cancer has a KRAS p.G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutation or a combination thereof.

11. 11. The method of any of claims 1 to 10, wherein the cancer has a KRAS p.G12C, G12D or G12V mutation or a combination thereof.

12. 12. The method of any of claims 1 to 11, wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, small intestine cancer, colon adenocarcinoma, rectal adenocarcinoma, small intestine adenocarcinoma, lung adenocarcinoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, plasma cell myeloma, gallbladder cancer, anaplastic thyroid cancer, ampullary carcinoma, cervical cancer, gastrointestinal neuroendocrine tumors, uterine endometrioid carcinoma, germ cell tumors, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumors, hepatobiliary cancer, histiocytosis, anal cancer, melanoma, mature b-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, salivary gland cancer, renal cell carcinoma, and bone cancer.

13. The method of any one of claims 1 to 12, wherein the cancer is KRAS inhibitor-resistant, KRAS inhibitor-sensitive, or KRAS inhibitor-low-sensitive, including intrinsic resistance, acquired resistance, or adaptive resistance.

14. 14. The method of claim 13, wherein the acquired KRAS inhibitor resistance is selected from G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N inhibitor resistance.

15. Acquired resistance to KRAS inhibitors has been reported for sotorasib (AMG510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, The method of claim 13, wherein the resistance is selected from resistance to JDQ443, MK-1084, MRTX1133, JAB-22000, RMC-9805 (RM-036), RMC-8839, JAB-23000, cobimetinib, RMC-6236, BBP-454, BI1701963, BAY-293, RMC-5845, BI-3406, SDGR5, or a pharmaceutically acceptable salt thereof.

16. A combination of a PIM kinase inhibitor and a KRAS inhibitor, wherein the PIM kinase inhibitor is one of those described in any one of claims 2 to 5, and the KRAS inhibitor is A combination according to any one of claims 6 to 8.

17. The combination according to claim 13, wherein the combination is for the treatment of a cancer according to any one of claims 9 to 12.

18. Use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the manufacture of a medicament for the treatment of cancer, wherein the PIM kinase inhibitor is as defined in any one of claims 2 to 5 and the KRAS inhibitor is Use according to any one of claims 6 to 8.

19. The use according to claim 18, wherein the cancer is one of the cancers according to any one of claims 10 to 15.

20. A pharmaceutical composition comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable additive, wherein the PIM kinase inhibitor is one according to any one of claims 2 to 5, and the KRAS inhibitor is A pharmaceutical composition according to any one of claims 6 to 8.

21. The pharmaceutical composition of claim 20, wherein the pharmaceutical composition is for treating a cancer according to any one of claims 10 to 15.

22. A kit comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable additive, wherein the PIM kinase inhibitor is one according to any one of claims 2 to 5, and the KRAS inhibitor is A kit according to any one of claims 6 to 8.

23. The kit of claim 22, wherein the kit is for treating a cancer according to any one of claims 10 to 15.

24. 1. A method for treating a cancer having a KRAS mutation in a human subject in need thereof, comprising administering a therapeutically effective amount of N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide or a pharmaceutically acceptable salt thereof; or N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide or a pharmaceutically acceptable salt thereof; or use of N-(5-((2S,5R,6S)-5-amino-6-fluorooxepan-2-yl)-1-methyl-1H-pyrazol-4-yl)-2-(2,6-difluorophenyl)thiazole-4-carboxamide or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating cancer harboring a KRAS mutation.

25. 25. The method or compound or use of claim 24, wherein the KRAS mutation is selected from a G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutation.

26. 25. The method or compound or use of claim 24 or 25, wherein the cancer is KRAS inhibitor resistant, including intrinsic, acquired or adaptive resistance.

27. 27. The method, compound, or use of any of claims 24 to 26, wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, small intestine cancer, colon adenocarcinoma, rectal adenocarcinoma, small intestine adenocarcinoma, lung adenocarcinoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, plasma cell myeloma, gallbladder cancer, anaplastic thyroid cancer, ampullary carcinoma, cervical cancer, gastrointestinal neuroendocrine tumors, uterine endometrioid carcinoma, germ cell tumors, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumors, hepatobiliary cancer, histiocytosis, anal cancer, melanoma, mature b-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, salivary gland cancer, renal cell carcinoma, and bone cancer.

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