Combination of PIM kinase inhibitors with KRAS inhibitors

Abstract

The present disclosure relates to a combination of a PIM kinase inhibitor and a KRAS inhibitor for use in the treatment of cancer in a subject in need thereof. Also provided are compositions or kits comprising the same. The disclosure also relates to PIM kinase inhibitors for use in the treatment of cancer with a KRAS mutation in a human subject in need thereof.

Description

Technical Field

[0001] The present invention provides a combination therapy comprising a PIM kinase inhibitor and a KRAS inhibitor for treating cancer. The present invention also relates to a pharmaceutical composition or a kit comprising a PIM kinase inhibitor and a KRAS inhibitor for treating cancer. The present invention also relates to a PIM kinase inhibitor for treating cancer having a KRAS mutation. Background Art

[0002] KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer (CRC), gallbladder cancer, thyroid cancer, and cholangiocarcinoma. Among KRAS mutations, the G12 codon (81%) has the highest mutation frequency, followed by G13 (14%) and Q61 (2%). KRAS mutation is 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 carcinoma (8.6%) (Kwan et al. J Exp Clin Cancer Res (2022) 41:27). Therefore, agents that block the proliferative signal transduction induced by oncogenic KRAS variants have received intense attention.

[0003] KRAS has been considered an undruggable target for decades, but in 2013, researchers identified a cryptic pocket adjacent to the mutant cysteine in the KRAS G12C protein, which is only shown in the GDP-bound form, ultimately providing a direct drug-binding site. Various attempts have been made to develop KRAS inhibitors. In May 2021, AMG510 (sotorasib) became the first FDA-approved therapy directly targeting KRAS mutant tumors. In June 2021, MRTX849 (adagrasib) received FDA breakthrough therapy designation. Other KRAS inhibitors are currently under approval, 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, BI 1701963, and BAY-293.

[0004] Although in many patients receiving KRAS G12CClinical benefit has been observed in patients treated with the inhibitors adagrasib or sotorasib, and acquired resistance to single-agent therapy eventually develops in most patients. Patients with KRAS mutant tumors have significantly worse outcomes and a poor prognosis. An effective combination therapy appears to be necessary to overcome this acquired resistance to direct KRAS G12C inhibitors. WO2020106647A2 discloses combinations of KRAS G12C inhibitors with carboplatin, anti-PD-1 inhibitors, MEK inhibitors, EGFR inhibitors, TOR inhibitors, SHP2 inhibitors, PI3K inhibitors or AKT inhibitors. JANE DELARTIGUE also discloses combinations of KRAS G12C inhibitors with pan-ERBB inhibitors, CDK4 / 6 inhibitors, SOS1 / pan-KRAS inhibitors (Jane de Lartigue. OncologyLive, Vol.23 / No.1, Volume 23, Issue 01.table).

[0005] In addition, due to a lack of clinical efficacy, some therapies targeting the MAPK pathway are prohibited for the treatment of KRAS mutant tumors. Additionally, due to the inhibition of MAPK signaling in normal cells, non-tumor or non-mutant selective therapies may introduce on-target toxicity. This may limit the utility of these agents in combination with the standard of care or immunotherapy. Accordingly, there is a significant unmet need for the development of tumor-selective therapies that do not cause damage to normal cells. To date, there has been no research on combination therapies of PIM inhibitors and KRAS inhibitors, particularly combinations of PIM inhibitors and KRAS G12C inhibitors, KRAS G12D inhibitors or KRAS G12V inhibitors. SUMMARY OF THE INVENTION

[0006] To overcome resistance to KRAS inhibitors during an extended treatment period, the present disclosure provides combination therapies comprising a PIM kinase inhibitor (PIMi) and a KRAS inhibitor. The present disclosure has found that resistance to KRAS inhibitors, particularly acquired resistance, can be reversed by co-treatment with a PIM kinase inhibitor. Additionally, the present disclosure has found that the combination of a PIM kinase inhibitor and a KRAS inhibitor exhibits superior efficacy compared to either single therapy and demonstrates synergy against KRAS mutant cancers. Further, the present disclosure has also found that the PIM inhibitor GDC-0570 can be used as a single active agent in multiple KRAS mutant cancers.

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

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

[0009] In another aspect, the present disclosure provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the treatment of cancer, such as KRAS mutant cancer or KRAS inhibitor-resistant cancer.

[0010] In another aspect, the present disclosure provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the preparation of a medicament for treating cancer (such as KRAS mutant cancer or KRAS inhibitor-resistant cancer).

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

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

[0013] In yet another aspect, the present disclosure provides a method for treating cancer having a KRAS mutation in a human subject in need thereof, which comprises 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 can be used as a single active agent in a variety of KRAS mutant cancers. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Shows the effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant CRC PDX model CRC024.

[0015] Figure 2 Shows the effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant NSCLC PDX model LUN055.

[0016] Figure 3 Shows the effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant NSCLC PDX model LUN156.

[0017] Figure 4Show the effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant NSCLC PDX model LUN2156-44.

[0018] Figure 5 Show the effects of GDC-0570 and GDC-6036 on tumor volume in the KRAS G12C mutant CRC PDX model CRC022.

[0019] Figure 6 Show the effects of GDC-0570 on tumor volume in the KRAS G12C mutant NSCLC PDX model LUN156.

[0020] Figure 7 Show the effects of GDC-0570 on tumor volume in the KRAS G12D mutant NSCLC PDX model LUN#137.

[0021] Figure 8 Show the effects of GDC-0570 and cobimetinib on tumor volume in the KRAS G12D mutant pancreatic PDX model PAN092.

[0022] Figure 9 Show the effects of GDC-0570 and MRTX1133 on tumor volume in the KRAS G12D pancreatic cancer model PAN092.

[0023] Figure 10 Show the effects of GDC-0570 and cobimetinib on tumor volume in the KRAS G12V mutant CRC PDX model. Detailed implementation

[0024] Definition

[0025] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0026] The terms "comprising", "including", "containing", "having included" and "having contained" are intended to specify the presence of the stated features, wholes, components or steps, but they do not preclude the presence or addition of one or more other features, wholes, components, steps or combinations thereof.

[0027] As used herein, the singular forms "a", "an" and "the" include both singular and plural referents unless the context clearly dictates otherwise.

[0028] The term "optional" or "optionally" means that the subsequent described event, circumstance or substitution may or may not occur, and the description includes both the case where the event or circumstance occurs and the case where it does not occur.

[0029] The terms "about" or "approximately" generally mean within 5% of a given value or range, or more preferably within 1%.

[0030] Numerical ranges expressed by endpoints include all values and fractions within the corresponding range, as well as the expressed endpoints.

[0031] The terms "treat", "treatment" and "treating" refer to a therapeutic treatment of a subject having cancer, which is intended to prevent or slow down (alleviate) an undesired physiological change or condition, such as the growth, development or spread of cancer. For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, reduction in the degree of disease, stabilization (i.e., not worsening) of the disease state, delay or slowing of disease progression, improvement or alleviation of the disease state, and remission (whether partial or complete). "Treatment" may also refer to an extension of survival as compared to expected survival in the absence of treatment.

[0032] The phrase "therapeutically effective amount" means an 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 has been shown to have an improvement in terms of (i), (ii) or (iii) as compared to monotherapy with a single agent. The therapeutically effective amount of the combination may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow down to some extent and preferably stop) the infiltration of cancer cells into surrounding organs; inhibit (i.e., slow down to some extent and preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent. To the extent that the combination can prevent growth and / or kill existing cancer cells, it can be cytostatic and / or cytotoxic. For cancer treatment, efficacy can be measured, for example, by assessing the time to disease progression (TTP) and / or determining the response rate (RR).

[0033] As used herein, the term "synergistic" refers to a therapeutic combination that is more effective than the additive effect of two or more single agents. The determination of the synergistic interaction between a PIM kinase inhibitor and a KRAS inhibitor can be based on the results obtained from the assays described herein. The combination therapy can provide "synergy" and be demonstrated to be "synergistic", i.e., the effect obtained when the active ingredients are used together is greater than the sum of the effects produced by the compounds used alone. In one example, a synergistic effect can be achieved when the active ingredients are: (1) co-formulated and administered or delivered simultaneously in a combined unit dose formulation; (2) delivered alternately or in parallel as separate formulations; or (3) delivered by some other regimen. When delivered by an alternating therapy, a synergistic effect can be obtained when, for example, the compounds are administered or delivered by different injections in separate syringes or by different oral dose sequences. Generally, during an alternating therapy, the effective doses of each active ingredient are administered sequentially, i.e., in sequence, while in a combination therapy, the effective doses of two or more active ingredients are administered together. Synergy can be evaluated by tumor growth inhibition. Specifically, the tumor volume growth trend is inhibited, preferably the tumor volume is significantly reduced, or the tumor completely regresses.

[0034] "Drug-resistant cancer" or "refractory cancer" are used interchangeably herein and refer to cancer that is unresponsive or less responsive to therapeutic treatment. Drug-resistant cancer can have intrinsic drug resistance, acquired drug resistance, or adaptive drug resistance. "Intrinsic drug resistance" or "primary drug resistance" means that the tumor lacks a response to the initial therapy. "Acquired drug resistance" refers to a tumor that initially responds to treatment but then recurs. "Adaptive drug resistance" refers to drug resistance induced by alterations in upstream or downstream or parallel pathway components of KRAS mutant cancers, which inevitably leads to a lack of efficacy, as well as recurrence and progression of these tumors. For example, "KRAS drug-resistant cancer" includes cancers that have acquired drug resistance to one or more KRAS inhibitors following treatment with one or more KRAS inhibitors, or can include cancers that have intrinsic drug resistance to one or more KRAS inhibitors, such as cancers with activated PIM kinases.

[0035] Drug-resistant cancer can have drug resistance at the start of treatment, or it can become drug-resistant during treatment.

[0036] As used herein, the term "sensitive cancer" refers to cancer that responds to a certain drug treatment and shows no progression. For example, sensitive cancer is sotorasib-sensitive KRAS G12C mutant colorectal cancer or sotorasib-sensitive KRAS G12C mutant NSCLC cancer. Administration of 100 mg / kg sotorasib to PDX models of these cancers results in >100% TGI.

[0037] As used herein, the term "less sensitive cancer" refers to cancers that progress during certain drug treatments. For example, a less sensitive cancer is a KRAS G12C mutant NSCLC cancer that is less sensitive to sotorasib. Administration of 100 mg / kg sotorasib to a PDX model of this cancer results in <100% TGI.

[0038] The term "combination" refers to simultaneous, separate, or sequential administration. When the administration is sequential or separate, the delay in administration of the second component should not, for example, result in loss of the beneficial effects of the combination.

[0039] 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 human male subject. In some embodiments, the subject is a human female subject. The terms "subject", "patient", and "individual" are also used interchangeably herein.

[0040] The phrase "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal being treated therewith.

[0041] The phrase "pharmaceutically acceptable salt" refers to pharmaceutically acceptable organic or inorganic salts of a compound. Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, hydrogen phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, galacturonate, formate, benzoate, glutamate, mesylate, "mesylate", esylate, besylate, tosylate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). A pharmaceutically acceptable salt may involve the inclusion of another molecule, such as an acetate ion, a succinate ion, or other counterions. The counterion can be any organic or inorganic moiety that stabilizes the charge on the parent compound. In addition, a pharmaceutically acceptable salt may have more than one charged atom in its structure. When multiple charged atoms are part of a pharmaceutically acceptable salt, there may be multiple counterions. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions. If the compound is a base, the desired pharmaceutically acceptable salt can 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, etc.) or with 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, pyranosyl acids (such as glucuronic acid or galacturonic acid), alpha-hydroxy acids (such as citric acid or tartaric acid), amino acids (such as aspartic acid or glutamic acid), aromatic acids (such as benzoic acid or cinnamic acid), sulfonic acids (such as p-toluenesulfonic acid or ethanesulfonic acid), etc.).Acids generally considered suitable for forming pharmaceutically useful or acceptable salts 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) 119; 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 (U.S. Food and Drug Administration, Washington, D.C., on its website). If the compound is an acid, the required pharmaceutically acceptable salts can be prepared by any suitable method, such as treating the free acid with an inorganic or organic base (such as an amine, e.g., a primary, secondary or tertiary amine), an alkali metal hydroxide or an alkaline earth metal hydroxide, etc. Exemplary examples of suitable salts include, but are not limited to, organic salts derived from amino acids (such as glycine and arginine), ammonia, primary amines, secondary amines 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.

[0042] All publications, published patent documents and patent applications cited herein are incorporated herein by reference to the extent that each individual publication, published patent document or patent application is specifically and individually indicated to be incorporated by reference.

[0043] 1. Exemplary PIM kinase inhibitors and KRAS inhibitors

[0044] The present disclosure generally relates to combinations of PIM kinase inhibitors and KRAS inhibitors as described herein, for example, for treating cancer.

[0045] 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 effective activity against PIM-1, PIM-2, and / or PIM-3 inhibitors. Exemplary PIM kinase inhibitors include, but are not limited to, AZD1208, LGH447, and the compounds disclosed in WO2014048939, US20110059961, or US20130079321 (such as GDC-0570, GNE-1571, GNE-5775, GDC-0339, and GNE-5652) and their pharmaceutically acceptable salts, the structures of which are provided below:

[0046]

[0047] In some embodiments, the PIM kinase inhibitor is selected from GDC-0570, GNE-1571, GNE-5775, GDC-0339, GNE-5652, and their pharmaceutically acceptable salts.

[0048] In some 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 its pharmaceutically acceptable salt. GDC-0570 is Compound 321 in WO2014048939.

[0049] In some 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 its pharmaceutically acceptable salt. GNE-1571 is Compound 322 in WO2014048939.

[0050] 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 its pharmaceutically acceptable salt. GNE-5775 is Compound 231 in WO2014048939.

[0051] In some embodiments, the PIM kinase inhibitor is GDC-0339, also known as 5-amino-N-(5-((4R,5R)-4-amino-5-fluorazepan-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.

[0052] 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.

[0053] In some embodiments, the KRAS inhibitor is 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 inhibitor, pan-KRAS inhibitor, KRAS-SOS1 interaction inhibitor or KRAS signaling inhibitor, and a pharmaceutically acceptable salt thereof.

[0054] In some embodiments, the KRAS G12C inhibitor is sotorasib (AMG 510), 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 a pharmaceutically acceptable salt thereof. Some structures of the KRAS G12C inhibitors are provided below:

[0055]

[0056]

[0057] In some embodiments, the KRAS G12CThe inhibitors are disclosed in WO2014152588, WO2015054572, WO2016049524, WO2016164675, WO2016168540, WO2017015562, WO2017058915, WO2017058807, WO2017058792, WO2017058902, WO2017087528, WO2017201161, WO2018064510, WO2018068017, WO2018119183, WO2018140600, WO2018140512, WO2018143315, WO2018206539, WO2018217651, WO2018218070, WO2019051291, WO2019099524, WO2019110751, WO2019137985, WO2019141250, CN111377918, CN112159405, CN112574199, WO2021155716, WO2021197499, WO2021249563 WO2022028346, WO2022037560, WO2022068921, WO2022111521 and WO2022135591.

[0058] In some embodiments, the KRAS G12D inhibitor is selected from MRTX1133, JAB-22000, RMC-9805 (RM-036) and pharmaceutically acceptable salts thereof.

[0059]

[0060] In some embodiments, the KRAS G13C inhibitor is selected from RMC-8839 and pharmaceutically acceptable salts thereof.

[0061] In some embodiments, the KRAS G12V inhibitor is selected from JAB-23000 and pharmaceutically acceptable salts thereof.

[0062] In some embodiments, the pan-KRAS inhibitor is selected from RMC-6236, BBP-454 and pharmaceutically acceptable salts thereof.

[0063] In some embodiments, the KRAS signaling inhibitor is a MEK inhibitor that inhibits mitogen-activated protein kinase 1 (MAP2K1 or MEK1) and a central component of the RAS / RAF / MEK / ERK signal transduction pathway. In some embodiments, the KRAS signaling inhibitor is the MEK inhibitor cobimetinib. Cobimetinib inhibits signaling downstream of all oncogenic KRAS (e.g., KRAS G12D , KRAS G12V ).

[0064]

[0065] In some embodiments, the KRAS-SOS1 interaction inhibitor is selected from BI 1701963, BAY-293, RMC-5845, BI-3406, SDGR5, and pharmaceutically acceptable salts thereof. In some embodiments, the KRAS-SOS1 interaction inhibitors are disclosed in WO2018115380, WO2019122129, WO2018172250, WO2016077793, and WO2022017339.

[0066] The PIM kinase inhibitor and the KRAS inhibitor can be in the form of isotopically labeled compounds that are the same as those described herein, but with one or more atoms replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. All isotopes of any specified atom or element are included within the scope of the compounds of the invention and their uses. Exemplary isotopes that can be incorporated into the compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 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., those labeled with 3 H and 14 C) can be used for compound and / or substrate tissue distribution assays. Tritiated ( 3 H) and carbon-14 ( 14C) Isotopes are useful because of their ease of preparation and detectability. In addition, substitution with heavier isotopes such as deuterium ( 2 H) can provide certain therapeutic advantages resulting from higher metabolic stability (e.g., increased in vivo half-life or reduced dose requirements), and can thus be preferred in some cases. Positron-emitting isotopes such as 15 O, 13 N, 11 C, and 18 F can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0067] In certain embodiments, the PIM kinase inhibitor is selected from GDC-0570, GNE-1571, GNE-5775, GDC-0339, and GNE-5652 and their pharmaceutically acceptable salts, and the KRAS G12C inhibitor is selected from sotorasib (AMG 510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI 1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and their pharmaceutically acceptable salts.

[0068] 2. Treatment of cancer

[0069] In another aspect, the present disclosure provides methods for treating cancer in a subject in need thereof, such as methods for treating KRAS mutant cancers or KRAS inhibitor-resistant cancers, which comprise administering to the subject a therapeutically effective amount of a PIM kinase inhibitor and a therapeutically effective amount of a KRAS inhibitor.

[0070] In another aspect, the present disclosure provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the treatment of cancer, such as KRAS mutant cancers or KRAS inhibitor-resistant cancers.

[0071] In another aspect, the present disclosure provides the use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the preparation of a medicament for treating cancer (e.g., KRAS mutant cancers or KRAS inhibitor-resistant cancers).

[0072] "Tumor" and "cancer" are used interchangeably herein and refer to a physiological condition in a mammal that is typically characterized by unregulated 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.

[0073] In some embodiments, the cancer includes, but is not limited to, lung cancer, colorectal cancer, pancreatic cancer, appendiceal 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 cancer, cervical cancer, gastroenteropancreatic neuroendocrine tumor, endometrioid carcinoma, germ cell tumor, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord stromal tumor, hepatobiliary duct cancer, histiocytosis, anal cancer, melanoma, mature B cell tumor, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland adenocarcinoma, breast cancer, renal cell cancer, and bone cancer.

[0074] In some embodiments, the cancer is a KRAS inhibitor-resistant cancer. In some embodiments, KRAS inhibitor-resistant cancers include intrinsic resistance, acquired resistance, or adaptive resistance. "KRAS-resistant cancer" includes cancers that have been previously treated with one or more KRAS inhibitors and have acquired resistance to one or more KRAS inhibitors, or may include cancers that are intrinsically resistant to one or more KRAS inhibitors.

[0075] In some embodiments, acquired KRAS resistance is induced by a KRAS inhibitor. In some embodiments, acquired KRAS resistance is induced by sotorasib (AMG 510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI 1823911, 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 a sotorasib (AMG 510)-resistant cancer or an adagrasib (MRTX849)-resistant cancer. The resistant cancer may be resistant at the start of treatment, or it may become resistant during treatment. In some embodiments, sotorasib resistance is induced by adagrasib.

[0076] In some embodiments, KRAS inhibitor-resistant cancers are selected from lung cancer, colorectal cancer, pancreatic cancer, appendiceal 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 cancer, cervical cancer, gastroenteric neuroendocrine tumor, endometrioid carcinoma, germ cell tumor, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumor, hepatobiliary duct cancer, histiocytosis, anal cancer, melanoma, mature B cell tumor, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, renal cell cancer, and bone cancer.

[0077] As described herein, the PIM kinase inhibitor and the KRAS inhibitor are each administered in a therapeutically effective amount in combination. In some embodiments, the administered amounts of the PIM kinase inhibitor and the KRAS inhibitor are more excellent and effective relative to either single therapy. In some embodiments, the administered amounts of the PIM kinase inhibitor and the KRAS inhibitor are synergistic for KRAS mutant cancers. In some embodiments, the KRAS mutant cancers have KRAS p.G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutations or combinations thereof. In some embodiments, the cancer has a KRAS p.G12C or G12D mutation or combination thereof.

[0078] In some embodiments, the weight ratio of the PIM kinase inhibitor to the KRAS inhibitor is from about 1:0.001 to 0.001:1, from about 1:0.01 to about 1:100, from about 1:0.1 to about 1:10, from about 5:1 to about 1:5, or from about 1:1 to about 1:5. In some 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, about 1:100. In some 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, about 1:100. Preferably, the KRAS inhibitor is selected from KRAS G12C inhibitors, KRAS G12V inhibitors, KRASG12D Inhibitor, KRAS G13C Inhibitor, KRAS G13D Inhibitor, KRAS Q61H Inhibitor, KRAS Q61L Inhibitor, KRAS Q61R Inhibitor, KRAS K117N Inhibitor, pan - KRAS inhibitor, KRAS - SOS1 interaction inhibitor or KRAS signaling inhibitor.

[0079] In some embodiments, the weight ratio of GDC - 0570 to the KRAS G12C 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 some embodiments, the weight ratio of the PIM kinase inhibitor to the KRAS G12C inhibitor 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. In some embodiments, the KRAS G12C inhibitor is sotorasib or RG6330 (GDC - 6036).

[0080] In some embodiments, the weight ratio of GDC - 0570 to the KRAS G12D 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 G12D 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, about 1:30, about 1:100. In some embodiments, the KRAS G12D inhibitor is MRTX1133 or the KRAS G12D signaling inhibitor cobimetinib. Cobimetinib is a MEK inhibitor and thus inhibits KRAS G12D signaling.

[0081] In some embodiments, the weight ratio of GDC - 0570 to the KRASG12V The weight ratio of the 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, GDC-0570 and KRAS G12V The weight ratio of the 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, about 1:100. In some embodiments, KRAS G12V The inhibitor is a KRAS G12V signal transduction inhibitor cobimetinib. Cobimetinib is a MEK inhibitor and thus inhibits KRAS G12V signal transduction.

[0082] In some embodiments, the weight ratio of GDC-0570 to the KRAS signal transduction 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 signal transduction 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, about 1:100. In some embodiments, the KRAS signal transduction inhibitor is cobimetinib. Cobimetinib is a MEK inhibitor and inhibits signal transduction downstream of all oncogenic KRAS (e.g., KRAS G12D , KRAS G12V ).

[0083] In some 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 some 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.

[0084] 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 may be administered in two or more administrations. The combination administration includes co - administration, using separate formulations, and sequential administration in either order, wherein preferably there is a period of time during which two (or all) of the active agents exert their biological activity simultaneously.

[0085] Suitable doses of any of the above co - administered agents are those currently in use and may be reduced due to the combined action (synergism) of the newly identified agent and other chemotherapeutic agents or treatments, for example to increase the therapeutic index or to mitigate toxicity or other side effects or consequences.

[0086] In further embodiments, the method may further include surgical treatment and / or radiotherapy. The amounts and relative timing of administration of the PIM kinase inhibitor, the KRAS inhibitor, and other pharmaceutically active chemotherapeutic agents are selected to achieve the desired combined therapeutic effect.

[0087] In some embodiments, the present disclosure provides methods for treating cancers having a KRAS mutation in a human subject in need thereof, the methods 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 some embodiments, the present disclosure provides GDC-0570 or a pharmaceutically acceptable salt thereof for use as a medicament for treating cancers having a KRAS mutation. In some embodiments, the present disclosure provides the use of GDC-0570 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancers having a KRAS mutation. In some embodiments, GDC-0570 can be used as a single active agent in a variety of KRAS mutant cancers. In some embodiments, the KRAS mutation is selected from the group consisting of G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutations. In some embodiments, the cancer is resistant to a KRAS inhibitor, the KRAS inhibitor resistance including intrinsic resistance, acquired resistance or adaptive resistance. In some embodiments, the cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendiceal 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 cancer, cervical cancer, gastroenteropancreatic neuroendocrine tumor, endometrioid carcinoma, germ cell tumor, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumor, intrahepatic cholangiocarcinoma, histiocytosis, anal cancer, melanoma, mature b-cell tumor, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, renal cell cancer and bone cancer. In some embodiments, the KRAS inhibitor-resistant cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendiceal 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 cancer, cervical cancer, gastroenteropancreatic neuroendocrine tumor, endometrioid carcinoma, germ cell tumor, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumor, intrahepatic cholangiocarcinoma, histiocytosis, anal cancer, melanoma, mature b-cell tumor, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, renal cell cancer and bone cancer.

[0088] 3. Combinations, Pharmaceutical Compositions and Kits

[0089] In another aspect, the present disclosure provides a combination comprising a PIM kinase inhibitor and a KRAS inhibitor. Preferably, the combination is for treating cancer, such as KRAS mutant cancer or KRAS inhibitor-resistant cancer.

[0090] In certain embodiments, the combination is provided as a single pharmaceutical composition together with a pharmaceutically acceptable excipient. In other embodiments, the combination is provided as two pharmaceutical compositions, one comprising a PIM kinase inhibitor and a pharmaceutically acceptable excipient, and the other comprising a KRAS inhibitor and a pharmaceutically acceptable excipient, and the two pharmaceutical compositions are administered in combination together.

[0091] 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 as described herein. A pharmaceutically acceptable excipient is inert. Non-limiting examples of pharmaceutically acceptable excipients include pharmaceutically acceptable polymers, water, NaCl, saline solution, common sucrose, common glucose, binders, fillers, disintegrants, lubricants, surfactants, coatings, sweeteners, flavorants, salt solutions, alcohols, oils, gelatin, carbohydrates, pigments, and the like. Such formulations may be sterile and, if desired, may be admixed with adjuvants such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts affecting osmotic pressure, buffers, colorants, and / or aromatic substances, and the like. Pharmaceutically acceptable excipients are described in the Handbook of pharmaceutical excipients, 8th edition, published by Pharmaceutical Press (2017) and the U.S. Food and Drug Administration Inactive Ingredient Database (July 2017), the disclosures of which are incorporated herein by reference.

[0092] Depending on the method used to administer the drug, the pharmaceutical composition can be packaged in a variety of ways. Generally, an article for dispensing includes a container that houses a pharmaceutical formulation in a suitable form. 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 an anti-tampering fitting to prevent inadvertent contact with the contents of the package. In addition, the container has a label that describes the contents of the container. The label may also include appropriate warnings.

[0093] Pharmaceutical compositions can be prepared for a variety of routes and types of administration. The pharmaceutical composition will be administered and dispensed in a manner consistent with good medical practice (i.e., amount, concentration, schedule, procedure, vehicle, and route of administration). Factors considered herein include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the dosing schedule, and other factors known to the practicing physician.

[0094] In certain embodiments, the pharmaceutical composition is formulated for oral delivery. Formulations of PIM kinase inhibitors and / or KRAS inhibitors suitable for oral administration can be prepared as discrete units, such as pills, hard or soft gelatin capsules (e.g., gelatin capsules), cachets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, syrups or elixirs, each containing a predetermined amount of the PIM kinase inhibitor and / or KRAS inhibitor. The amounts of the PIM kinase inhibitor and KRAS inhibitor of the compound can be formulated as a pill, capsule, solution or suspension as a combination preparation. Alternatively, the PIM kinase inhibitor and KRAS inhibitor can be formulated separately as a pill, capsule, solution or suspension for alternate administration.

[0095] In an embodiment, the pharmaceutical composition is a solid dosage form for oral administration, such as a tablet, capsule or pill. In an embodiment, the solid dosage form is a tablet.

[0096] The dose can be administered once daily (QD), twice daily (BID) or more frequently, depending on the pharmacokinetic (PK) and pharmacodynamic (PD) properties, including the absorption, distribution, metabolism and excretion of the specific compound. In addition, toxicity factors can affect the dose and dosing regimen. When administered orally, the pill, capsule or tablet can be taken twice daily, daily or at a lower frequency, such as weekly or biweekly or triweekly, for a specified period of time. This regimen can be repeated for multiple treatment cycles.

[0097] In another aspect, the present disclosure provides a kit comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable excipient.

[0098] The pharmaceutical kit may further comprise a label or package insert on or associated with the container. The term "package insert" is used to refer to the instructions which are usually included in the commercial packaging of a therapeutic product and which contain information about indications, usage, dosage, administration, contraindications and / or warnings regarding the use of such therapeutic product. Suitable containers include, for example, bottles, vials, syringes, blister packs, etc. The container may be made of various materials such as glass or plastic. The container may contain a compound effective in treating a disorder or a pharmaceutically acceptable salt thereof or a formulation thereof, and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial with a stopper pierceable by a hypodermic needle). At least one active agent in the composition is a PIM kinase inhibitor and / or a compound of a PIM kinase inhibitor. The label or package insert indicates that the composition is for treating the selected disorder. Alternatively or additionally, the article may further comprise a second container which contains a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution and glucose solution. It may also include other materials required from a commercial and user perspective, including other buffers, diluents, filters, needles and syringes.

[0099] The pharmaceutical kit may further comprise a PIM kinase inhibitor and instructions for administration of the PIM kinase inhibitor. For example, if the kit contains a first composition containing a PIM kinase inhibitor and a PIM kinase inhibitor, the kit may further comprise instructions for co-administering, sequentially or separately, the PIM kinase inhibitor and a KRAS inhibitor to a subject in need thereof.

[0100] In another case, the pharmaceutical kit is adapted to deliver a PIM kinase inhibitor and / or a solid oral form of a PIM kinase inhibitor, such as a tablet or a capsule. Such a kit preferably comprises a number of unit doses. Such a kit may include a card having the doses arranged in the order of their 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. If desired, a memory aid may be provided, such as in the form of numbers, letters or other markings or with a calendar insert, indicating the dates on which these doses may be administered in the treatment regimen.

[0101] The pharmaceutical kit may comprise (a) a first container containing a PIM kinase inhibitor therein; and (b) a second container containing a KRAS inhibitor therein. Alternatively or additionally, the kit may further comprise a third container which contains a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution and glucose solution. It may also include other materials required from a commercial and user perspective, including other buffers, diluents, filters, needles and syringes.

[0102] When the kit contains a PIM kinase inhibitor and a composition of a PIM kinase inhibitor, the kit may contain containers for holding the separate compositions, such as separate bottles or separate foil packages. However, the separate compositions may also be contained within a single undivided container. Typically, the kit includes instructions for administering the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (e.g., oral and parenteral), at different dosing intervals, or when the prescribing physician desires to titrate the individual components of the combination.

[0103] Examples

[0104] The present disclosure is further illustrated and explained by reference to the following examples. It should be noted that the following examples are illustrative only and are not intended to limit the scope of the present disclosure in any way.

[0105] Various PDX models of the present disclosure were established at GenenDesign Co., Ltd (Shanghai, China). The various PDX models used in the examples are summarized in Table 1.

[0106] Table 1. List of PDX models

[0107]

[0108] Example 1. Combination study of GDC-0570 and sotorasib in a patient-derived xenograft (PDX) model CRC024 of sotorasib-sensitive KRAS G12C mutant colorectal cancer

[0109] This study aimed to evaluate the in vivo combination antitumor efficacy of GDC-0570 and its combination with sotorasib in a patient-derived xenograft (PDX) model CRC024 of KRAS G12C mutant colorectal cancer (CRC). The CRC024 PDX model is a sotorasib-sensitive model derived from a 75-year-old Chinese female CRC cancer patient. The KRAS G12C mutation in the CRC PDX model was confirmed by whole exome sequencing and PCR sequencing. Tumor-bearing mice were divided into 4 groups, including a vehicle group, a 300 mg / kg GDC-0570 group, a 100 mg / kg sotorasib group, and a 300 mg / kg GDC-0570 plus 100 mg / kg sotorasib group. The dosing solution was administered orally daily. The study duration was 27 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0110] GDC-0570 vehicle: 0.5% methylcellulose (MC) and 0.2% Tween-80 (MCT), maintained at 2 - 8 °C.

[0111] GDC-0570 Administration Solution: Weigh the required amount of GDC-0570 powder and mix it with an appropriate amount of 0.5% MC 0.2% Tween 80 solution in a container to make the GDC-0570 concentration 60 mg / ml. Mix it by vortexing and sonication until it becomes homogeneous. Store the administration suspension at 2 - 8 °C for up to one week.

[0112] Sotorasib Vehicle: 50% w / w polyethylene glycol 400 (PEG400) + 50% w / w propylene glycol (PG).

[0113] Sotorasib Administration Solution: Weigh the required amount of sotorasib powder into a container. Then add the 50% / 50% PEG400 / PG mixture to the container to make the sotorasib concentration 20 mg / ml. Add 1N hydrochloric acid solution to the same container at a final concentration of 0.39%. Disperse sotorasib by vortexing for 5 minutes, followed by sonication for 10 minutes until it is completely dissolved. Store the administration solution at 2 - 8 °C for up to one week.

[0114] Measurement and Calculation

[0115] Measure the tumor size twice a week, and measure the body weight daily before administration. Observe the clinical signs daily. Euthanize all animals on the 27th day after tumor size calibration. After the animals are euthanized, collect the tumor samples.

[0116] Tumor Volume (TV) = (Length × Width 2 ) / 2

[0117] Relative Tumor Volume (RTV) = TV f / TV 0 where TV 0 and TV f are the tumor volumes measured on day 0 and day 27 respectively;

[0118] Tumor Growth Inhibition T / C Ratio (%) = (RTV of treatment group / RTV of vehicle control group) × 100%;

[0119] Tumor Growth Inhibition Rate (TGI)

[0120] TGI = [1 - (TVt f - TVt 0 ) / (TVc f - TVc 0 )] × 100%

[0121] TVt f is the group average tumor volume (TV) of the treatment group on the final treatment day

[0122] TVt 0is the group-average TV of the treatment group on day 0 of treatment

[0123] TVc f is the group-average TV of the control group on the final treatment day

[0124] TVc 0 is the group-average TV of the control group on day 0 of treatment

[0125] Tumor regression percentage (% regression) = 100 × (TV 0 - TV f ) / TV 0

[0126] TV 0 is the group-average TV measured in the same group but on day 0 of treatment.

[0127] TV f is the group-average TV measured in the same group but on the final treatment day.

[0128] Percentage change in body weight (% BWC) = (BWc - BWi) / BWi × 100%, where "c" refers to the current value, "i" represents the initial value, and "BW" refers to body weight.

[0129] Data analysis

[0130] Plot the tumor growth curve with tumor volume on the Y-axis and time on the X-axis; plot the body weight change curve with animal body weight on the Y-axis and time on the X-axis. Analyze the tumor volume and percentage change in body weight data using the one-way analysis of variance (One Way-ANOVA) method, and then perform a significance test using the Bartlett test (p < 0.05).

[0131] Criteria for evaluating the tumor growth inhibitory effect of the test article by the tumor growth inhibition TGI rate:[[]]END]]

[0132] Significant tumor inhibitory effect: TGI (%) > 60% and P < 0.05;

[0133] Non-significant tumor inhibitory effect: TGI (%) ≤ 60% or P > 0.05

[0134] Study results

[0135] Tables 2, 3, 4, and Figure 1 show the effects of GDC-0570 and sotorasib on tumor growth in the KRAS G12C mutant CRC PDX model CRC024.

[0136] Under the research conditions of the current KRAS G12C mutant CRC PDX model, the single-therapy groups of GDC-0570 (300 mg / kg), sotorasib (100 mg / kg), and the combination treatment group of GDC-0570 (300 mg / kg) plus sotorasib (100 mg / kg) all showed significant tumor suppression effects. In addition, the combination group of GDC-0570 and sotorasib showed superior efficacy compared to either single-therapy treatment. In addition, the combination group of GDC-0570 and sotorasib showed efficacy superior to single-therapy treatment.

[0137] Mice in all dose groups did not experience severe weight loss (defined as a weight loss of more than 20%), and there were no other abnormalities throughout the study.

[0138] Table 2: Effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant CRC PDX model CRC024

[0139]

[0140] “ / ” indicates not applicable

[0141] Table 3: Summary of efficacy relative to vehicle in the KRAS G12C mutant CRC PDX model CRC024

[0142]

[0143] “ / ” indicates not applicable;

[0144] P < 0.05 means statistically significant.

[0145] Table 4: Combination efficacy relative to single therapy in the KRAS G12C mutant CRC PDX model CRC024 Summary of the method

[0146]

[0147] P < 0.05 means statistically significant.

[0148] Example 2. Combination study of GDC-0570 and sotorasib in the less sensitive KRAS G12C mutant NSCLC cancer patient-derived xenograft (PDX) model LUN055

[0149] This study aimed to evaluate the in vivo combination antitumor efficacy of GDC-0570 and its combination with sotorasib in the LUN055 patient-derived xenograft (PDX) model of KRAS G12C mutant NSCLC. The LUN055 PDX model is a sotorasib-insensitive model derived from a 60-year-old Chinese male NSCLC patient. The KRAS G12C mutation in the NSCLC PDX model was confirmed by whole-exome sequencing and PCR sequencing. Tumor-bearing mice were divided into 4 groups, including a vehicle group, a GDC-0570 group at 300 mg / kg, a sotorasib group at 100 mg / kg, and a group of 300 mg / kg GDC-0570 plus 100 mg / kg sotorasib. The dosing solutions were administered orally daily. The study duration was 28 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0150] The GDC-0570 vehicle, GDC-0570 dosing solution, sotorasib vehicle, sotorasib dosing solution, measurement and calculation, data analysis were the same as in Example 1.

[0151] Study Results

[0152] Table 5-7 and Figure 2 showed the effects of GDC-0570 and sotorasib on tumor growth in the KRAS G12C mutant NSCLC PDX model LUN055.

[0153] Under the study conditions of the current KRAS G12C mutant NSCLC PDX model, the single-agent treatment groups of GDC-0570 (300 mg / kg) and sotorasib (100 mg / kg) did not show significant tumor growth inhibition. In contrast, the combination of 300 mg / kg GDC-0570 plus 100 mg / kg sotorasib treatment showed significant tumor suppression. In addition, the GDC-0570 and sotorasib combination group showed superior efficacy compared to single-agent treatment.

[0154] Mice in all dose groups did not experience severe weight loss (defined as more than 20% weight loss), and there were no other abnormalities throughout the study.

[0155] Table 5: Effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant NSCLC PDX model LUN055

[0156]

[0157] “ / ” indicates not applicable

[0158] Table 6: Summary of Efficacy Relative to Vehicle in the KRAS G12C-Mutant NSCLC PDX Model LUN055

[0159]

[0160] “ / ” indicates not applicable;

[0161] P < 0.05 indicates statistical significance.

[0162] Table 7: Relative Combination Efficacy in the KRAS G12C-Mutant NSCLC PDX Model LUN055 Summary of monotherapy

[0163]

[0164] P < 0.05 indicates statistical significance.

[0165] Example 3. In the Sotorasib-Sensitive KRAS G12C-Mutant NSCLC Patient-Derived Xenograft (PDX) Model LUN156, the Combination of GDC-0570 and Sotorasib Prevented Tumor Regrowth after 28 Days of Treatment

[0166] This study aimed to evaluate the in vivo antitumor efficacy of GDC-0570 and sotorasib in the NSCLC PDX model LUN156. The LUN156 PDX model is a sotorasib-sensitive model derived from a 73-year-old Chinese male NSCLC patient. The KRAS G12C mutation in LUN156 was confirmed by whole-exome sequencing and PCR sequencing.

[0167] Tumor-bearing mice were divided into 4 groups, including a vehicle group, a 300 mg / kg GDC-0570 group, a 100 mg / kg sotorasib group, and a 300 mg / kg GDC-0570 plus 100 mg / kg sotorasib group. Administration was stopped after day 28, and tumor growth was continuously observed until day 45. Tumor volume was measured twice a week. Body weight was measured daily before administration.

[0168] The GDC-0570 vehicle, GDC-0570 dosing solution, sotorasib vehicle, sotorasib dosing solution, measurements and calculations, data analysis were similar to Example 1.

[0169] Study Results

[0170] Tables 8 - 11 and Figure 3 Show the effects of GDC-0570 and sotorasib on tumor growth in the sotorasib-sensitive KRAS G12C-mutant NSCLC PDX model.

[0171] Compared with the vehicle group, the single-agent treatment group of GDC-0570 at 300 mg / kg, the single-agent treatment group of sotorasib at 100 mg / kg, and their combination treatment all showed significant tumor growth inhibition (Tables 8 - 11). The combination group of GDC-0570 and sotorasib showed superior efficacy to the single-agent treatment of GDC-0570, which was statistically significant (Table 10). Although based on the p-value during the 28-day dosing period, the difference in TGI between the combination group and the sotorasib single-agent group was not statistically significant (Table 10), only 1 tumor showed complete regression in the sotorasib single-agent group, while all (5 / 5) tumors in the combination group completely regressed by day 28. In addition, after dosing was stopped on day 28, all (5 / 5) tumors in the GDC-0570 and sotorasib single-agent groups experienced regrowth, while only 1 small tumor reappeared in the combination group, which maintained a minimal volume until day 45. Therefore, the combination of GDC-0570 and sotorasib prevented the regrowth of tumors that occurred with sotorasib single-agent treatment after the end of dosing. In summary, the combination group of GDC-0570 and sotorasib showed excellent efficacy compared to single-agent treatment.

[0172] Mice in all dose groups did not experience severe weight loss (defined as a weight loss of more than 20%), and there were no other abnormalities throughout the study.

[0173] Table 8: Effects of GDC-0570 and sotorasib on tumor volume in the KRAS G12C mutant NSCLC PDX model LUN156

[0174]

[0175] “ / ” indicates not applicable

[0176] Table 9: Summary of efficacy relative to vehicle in the KRAS G12C mutant NSCLC PDX model LUN156

[0177]

[0178] “ / ” indicates not applicable;

[0179] P < 0.05 means statistically significant.

[0180] Table 10: Summary of combination efficacy relative to single-agent treatment in the KRAS G12C mutant NSCLC PDX model LUN156

[0181]

[0182] P < 0.05 means statistically significant.

[0183] Table 11: Summary of tumor regrowth in the combination group relative to the monotherapy group in the KRAS G12C mutant NSCLC PDX model LUN156

[0184]

[0185] "CR" indicates complete regression

[0186] Example 4. Combination study of GDC-0570 and sotorasib in the KRAS G12C NSCLC model LUN2156-44 with acquired resistance to sotorasib

[0187] This study aimed to evaluate the in vivo combination antitumor efficacy of GDC-0570 and its combination with sotorasib in a patient-derived xenograft (PDX) model LUN2156-44 of KRAS G12C mutant NSCLC. The LUN2156-44 PDX model is a sotorasib-resistant model derived from the LUN156 PDX model. The KRAS G12C mutation in LUN2156-44 was confirmed by whole exome sequencing and PCR sequencing. Tumor-bearing mice were divided into 6 groups, including a vehicle group, a group of 150 mg / kg GDC-0570, a group of 100 mg / kg sotorasib, a group of 50 mg / kg GDC-0570 plus 100 mg / kg sotorasib, a group of 100 mg / kg GDC-0570 plus 100 mg / kg sotorasib, and a group of 150 mg / kg GDC-0570 plus 100 mg / kg sotorasib. The dosing solution was administered orally daily. The study duration was 28 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0188] The GDC-0570 vehicle, GDC-0570 dosing solution, sotorasib vehicle, sotorasib dosing solution, measurement and calculation, data analysis were similar to those in Example 1.

[0189] Study results

[0190] Tables 12 - 14 and Figure 4 show the effects of GDC-0570 and sotorasib on tumor growth in the KRAS G12C mutant NSCLC PDX model LUN2156-44.

[0191] Compared with the sotorasib-sensitive PDX model LUN156 on day 28, the sotorasib-resistant PDX model LUN2156-44 reduced the single-agent activity of sotorasib from complete regression to 88% TGI, and reduced the single-agent activity of GDC-0570 from stasis to <50% TGI.

[0192] Measurements of tumor volume on day 28 showed that treatment with 150 mg / kg of GDC-0570 had 38% tumor growth inhibition (TGI) and 71% T / C ratio (%). Treatment with 100 mg / kg of sotorasib had 88% TGI and 33% T / C ratio (%). The combination of 50 mg / kg of GDC-0570 and 100 mg / kg of sotorasib showed 92% tumor regression and 16% T / C ratio (%). The combination of 100 mg / kg of GDC-0570 and 100 mg / kg of sotorasib showed 95% tumor regression and 1% T / C ratio (%). The combination of 150 mg / kg of GDC-0570 and 100 mg / kg of sotorasib showed 98% tumor regression and a T / C ratio (%) close to 0%. Treatment with 150 mg / kg GDC-0570 monotherapy showed no significant TGI compared to the vehicle group. The 100 mg / kg sotorasib monotherapy group and its combinations with 3 different doses of GDC-0570 showed significant tumor suppression. In addition, the combinations of 100 mg / kg of sotorasib with 50 mg / kg, 100 mg / kg, or 150 mg / kg of GDC-0570 showed superior efficacy compared to 100 mg / kg sotorasib monotherapy treatment. Complete regression was observed even at 50 mg / kg GDC-0570 among all tested combination doses. This combination reduced the dose of GDC-0570 required to achieve complete regression and produced a strong synergistic effect.

[0193] Mice in all dose groups did not experience severe weight loss (defined as weight loss exceeding 20%) and had no other abnormalities throughout the study.

[0194] Table 12: Effects of GDC-0570 and sotorasib on tumor volume in NSCLC PDX model LUN2156-44

[0195]

[0196] “ / ” indicates not applicable

[0197] Table 13: Summary of efficacy relative to vehicle in PDX model LUN2156-44

[0198]

[0199] “ / ” indicates not applicable;

[0200] P < 0.05 means statistically significant.

[0201] Table 14: Summary of combination efficacy relative to monotherapy in PDX model LUN2156-44

[0202]

[0203] P < 0.05 indicates statistical significance.

[0204] Example 5. Combinatorial study of GDC-0570 and GDC-6036 in the patient-derived xenograft (PDX) model CRC022 of KRAS G12C mutant colorectal cancer

[0205] This study aimed to evaluate the in vivo combinatorial antitumor efficacy of GDC-0570 and its combination with GDC-6036 in the patient-derived xenograft (PDX) model CRC022 of KRAS G12C mutant CRC. This PDX model was derived from a 49-year-old Chinese female CRC cancer patient. The KRAS G12C mutation in CRC#022 was confirmed by whole exome sequencing and PCR sequencing. Tumor-bearing mice were divided into 4 groups, including a vehicle group, a 100 mg / kg GDC-6036 QD group, a 300 mg / kg GDC-0570 QD group, and a group of 300 mg / kg GDC-0570 plus 100 mg / kg GDC-6036. The study lasted for 27 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0206] The GDC-0570 vehicle, GDC-0570 dosing solution, measurements and calculations, data analysis were similar to Example 1.

[0207] GDC-6036 vehicle: 0.5% methylcellulose (Methocel)

[0208] GDC-6036 dosing solution: Weigh an appropriate amount of GDC-6036 into a container. Add an appropriate volume of 0.5% methylcellulose to the container to prepare 20 mg / ml GDC-6032. Vortex and sonicate repeatedly (>1 hour) to obtain a homogeneous solution. The dosing solution was stored at 2 - 8 °C for up to one week.

[0209] Study results

[0210] Table 15-16 and Figure 5 show the effects of GDC-0570 and GDC-6036 on tumor growth in the KRAS G12C mutant colorectal PDX model.

[0211] On the 27th day of tumor volume measurement, the 100 mg / kg GDC-6036 QD group had 78% tumor growth inhibition (TGI) and 30% T / C ratio (%). The 300 mg / kg GDC-0570 QD group had 79% tumor growth inhibition (TGI) and 29% T / C ratio (%). The combination of 300 mg / kg of GDC-0570 plus 100 mg / kg of GDC-6036 had 99% tumor growth inhibition (TGI) and 11% T / C ratio (%).

[0212] Compared with the vehicle group, all single-agent groups and combination groups showed significant tumor suppression. The GDC-0570 single-agent group showed moderate to strong tumor suppression (70 - 80% TGI). The combination treatment of GDC-0570 and GDC-6036 showed superior efficacy compared to the single-agent treatment of GDC-0570 or GDC-6036.

[0213] Mice in all dose groups did not experience severe weight loss (defined as more than 20% body weight loss), and there were no other abnormalities throughout the study. The combination of GDC-0570 and GDC-6036 was well tolerated in mice (<5% body weight loss).

[0214] Table 15: Effect of GDC-0570 combination on tumor volume in CRC PDX model CRC022

[0215]

[0216] “ / ” indicates not applicable

[0217] Table 16: Summary of efficacy relative to vehicle in PDX model CRC022

[0218]

[0219] “ / ” indicates not applicable;

[0220] P < 0.05 means statistically significant.

[0221] Example 6. Dose-dependent single-agent activity study of GDC-0570 in KRAS G12C mutant NSCLC PDX model LUN156

[0222] This study aimed 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 5 dose groups, including a vehicle group, and groups of 50 mg / kg, 100 mg / kg, 200 mg / kg, and 300 mg / kg of GDC-0570. The dosing solution was administered orally daily. The study duration was 21 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0223] The GDC-0570 vehicle, measurements and calculations, data analysis were similar to those in Example 1.

[0224] GDC-0570 dosing solution: The 60 mg / ml GDC-0570 dosing solution of 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 one week.

[0225] Study Results

[0226] Table 17 - 18 and Figure 6 show the effect of GDC-0570 on tumor growth in the KRAS G12C mutant NSCLC PDX model LUN156.

[0227] Under the current study conditions in the LUN156 model, GDC-0570 at dose levels of 50, 100, 200, and 300 mg / kg all showed 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 weight loss exceeding 20%) or other abnormalities throughout the study, indicating that all doses of GDC-0570 were well tolerated.

[0228] Table 17: Effect of GDC-0570 on tumor volume in the NSCLC PDX model LUN156

[0229]

[0230] “ / ” indicates not applicable

[0231] Table 18: Effect of GDC-0570 on T / C ratio and TGI in the NSCLC PDX model LUN156

[0232]

[0233] “ / ” indicates not applicable;

[0234] P < 0.05 means statistically significant.

[0235] Example 7. Dose-Dependent Single Agent Activity Study of GDC-0570 in the KRAS G12D Mutation NSCLC PDX Model LUN#137

[0236] This study aimed to evaluate the in vivo anti-tumor 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. The KRAS G12D mutation in LUN#137 was confirmed by whole exome sequencing and PCR sequencing.

[0237] Tumor-bearing mice were divided into 5 dose groups, including a vehicle group, a GDC-0570 group at 50 mg / kg, a GDC-0570 group at 100 mg / kg, a GDC-0570 group at 200 mg / kg, and a GDC-0570 group at 300 mg / kg. The dosing solution was administered orally daily. The study duration was 22 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0238] The GDC-0570 vehicle, measurements and calculations, data analysis were similar to Example 1. The GDC-0570 dosing solution was similar to Example 6.

[0239] Study Results

[0240] Table 19-20 and Figure 7 show the effect of GDC-0570 on tumor growth in the KRAS G12D mutation NSCLC PDX model LUN#137.

[0241] Under the current study conditions of the LUN#137 model, GDC-0570 showed dose-dependent single agent anti-tumor activity, and significant tumor inhibition was observed at the 200 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 weight loss exceeding 20%) or other abnormalities throughout the study, indicating that all doses of GDC-0570 were well tolerated.

[0242] Table 19: Effect of GDC-0570 on Tumor Volume in the NSCLC PDX Model LUN#137

[0243]

[0244] “ / ” indicates not applicable

[0245] Table 20: Effect of GDC-0570 on T / C Ratio and TGI in the NSCLC PDX Model LUN#137

[0246]

[0247] “ / ” indicates not applicable;

[0248] P < 0.05 indicates statistical significance.

[0249] Example 8. KRAS G12D Combination study of GDC-0570 and cobimetinib in the pancreatic cancer model PAN092

[0250] This study aimed to evaluate the in vivo combination antitumor efficacy of GDC-0570 and its combination with the MEK inhibitor cobimetinib in the KRAS G12D-mutant patient-derived xenograft (PDX) model PAN092 of pancreatic cancer. This PDX model was derived from a 65-year-old Chinese male pancreatic cancer patient. The KRAS G12D mutation in PAN092 was confirmed by whole exome sequencing and PCR sequencing. Tumor-bearing mice were randomly assigned to treatment groups of 5 mice per group, including a vehicle group, a cobimetinib group at 2.5 mg / kg QD, a GDC-0570 group at 300 mg / kg QD, and a group of 300 mg / kg of GDC-0570 plus 2.5 mg / kg of cobimetinib. The study duration was 27 days. Tumor volume was measured twice a week. Body weight was measured daily before dosing.

[0251] The GDC-0570 vehicle, GDC-0570 dosing solution, measurement and calculation, data analysis were similar to those in Example 1.

[0252] Cobimetinib vehicle: 0.5% methylcellulose / 0.2% Tween-80 (MCT)

[0253] Cobimetinib dosing solution: Weigh an appropriate amount of cobimetinib into a container. Add an appropriate volume of MCT to the container to prepare 0.5 mg / ml of cobimetinib. Vortex and sonicate repeatedly to obtain a homogeneous solution. The dosing solution is stored at 2 - 8 °C for up to one week.

[0254] Study results

[0255] Table 21 - 23 and Figure 8 show the effects of GDC-0570 and cobimetinib on tumor growth in the KRAS G12D-mutant pancreatic PDX model PAN092.

[0256] On the 27th day, the measurement results of tumor volume showed that cobimetinib in the 2.5 mg / kg QD group had 65% tumor growth inhibition (TGI) and a T / C ratio (%) of 47%. The 300 mg / kg QD GDC-0570 group had 55% tumor growth inhibition (TGI) and a T / C ratio (%) of 56%. The combination of 300 mg / kg of GDC-0570 plus 2.5 mg / kg of cobimetinib had >100% tumor growth inhibition (TGI) and a T / C ratio (%) of 19%.

[0257] Compared with the vehicle group, all single-agent groups and combination groups showed significant tumor suppression. The cobimetinib single-agent group showed a moderate tumor suppression effect (65% TGI). The GDC-0570 single-agent group also showed a moderate tumor suppression effect (55% TGI). The combination treatment of GDC-0570 and cobimetinib showed significantly superior efficacy compared to the single-agent treatment of GDC-0570 or cobimetinib.

[0258] Table 21: Effect of GDC-0570 combination on tumor volume in PC PDX model PAN092

[0259]

[0260] “ / ” indicates not applicable

[0261] Table 22: Summary of efficacy relative to vehicle in PDX model PAN092

[0262]

[0263] “ / ” indicates not applicable; P < 0.05 means statistically significant.

[0264] Table 23: Summary of efficacy among treatment groups in PDX model PAN092

[0265]

[0266] P < 0.05 means statistically significant.

[0267] Example 9. KRAS G12D Combination study of GDC-0570 and MRTX1133 in pancreatic cancer model PAN092

[0268] This study aimed to evaluate the in vivo combined anti-tumor efficacy of GDC-0570 and its combination with the KRAS G12D inhibitor MRTX1133 in the KRAS G12D mutant patient-derived xenograft (PDX) model PAN092 of pancreatic cancer. This PDX model was derived from a 65-year-old Chinese male pancreatic cancer patient. The KRAS G12D mutation in PAN092 was confirmed by whole exome sequencing and PCR sequencing. Tumor-bearing mice were randomly divided into treatment groups of 5 mice each, including a vehicle group, an MRTX1133 group at 10 mg / kg QD, a GDC-0570 group at 300 mg / kg QD, and a group of 300 mg / kg of GDC-0570 plus 10 mg / kg of MRTX1133. The study lasted for 27 days. Tumor volumes were measured twice a week. Body weights were measured daily before dosing.

[0269] The GDC-0570 vehicle, the GDC-0570 dosing solution, the measurements and calculations, and the data analysis were similar to those in Example 1.

[0270] MRTX1133 vehicle: 10% Captisol in 50 mM citrate buffer, pH 5.0

[0271] MRTX1133 dosing solution: Weigh an appropriate amount of MRTX1133 into a container. Add an appropriate volume of the vehicle to the container to prepare a 2 mg / ml MRTX1133 formulation. Vortex and sonicate repeatedly (>1 hour) to obtain a homogeneous solution. The dosing solution was stored at 2 - 8°C for up to one week.

[0272] Study results

[0273] Table 24 - 25 and Figure 9 show the effects of GDC-0570 and MRTX1133 on tumor growth in the KRAS G12D mutant pancreatic PDX model.

[0274] Tumor volume measurement results on day 27 showed that MRTX1133 in the 10 mg / kg QD group had 74% tumor growth inhibition (TGI) and 40% T / C ratio (%). The 300 mg / kg QD GDC-0570 group had 55% tumor growth inhibition (TGI) and 56% T / C ratio (%). The combination of 300 mg / kg of GDC-0570 plus 10 mg / kg of MRTX1133 had 89% tumor growth inhibition (TGI) and 27% T / C ratio (%).

[0275] Compared with the vehicle group, all single-agent groups and combination groups showed significant tumor suppression. The GDC-0570 and MRRTX1133 single-agent groups showed weak to moderate tumor suppression (74% and 55% TGI, respectively). The combination treatment of GDC-0570 and MRRTX1133 showed efficacy superior to that of GDC-0570 single-agent treatment.

[0276] Table 24: Effect of GDC-0570 combination on tumor volume in the PC PDX model PAN092

[0277]

[0278] “ / ” indicates not applicable

[0279] Table 25: Summary of efficacy relative to vehicle in the PDX model PAN092

[0280]

[0281] “ / ” indicates not applicable; P < 0.05 means statistically significant.

[0282] Example 10. Combination study of GDC-0570 and cobimetinib in the KRAS G12V mutant colorectal cancer PDX model CRC051

[0283] This study aimed to evaluate the in vivo combination anti-tumor efficacy of GDC-0570 and its combination with cobimetinib in the KRAS G12V mutant colorectal patient-derived xenograft (PDX) model CRC051. This PDX model was from a Chinese colorectal cancer patient. The KRAS G12V mutation in CRC051 was confirmed by whole exome sequencing and PCR sequencing. Tumor-bearing mice were divided into 4 groups, including a vehicle group, a cobimetinib group at 2.5 mg / kg QD, a GDC-0570 group at 300 mg / kg QD, and a group of 300 mg / kg of GDC-0570 plus 2.5 mg / kg of cobimetinib. The study duration was 27 days. Tumor volume was measured twice a week. Body weight was measured daily before drug administration.

[0284] The GDC-0570 vehicle, GDC-0570 dosing solution, measurement and calculation, data analysis were the same as in Example 1.

[0285] The cobimetinib vehicle and cobimetinib dosing solution were the same as in Example 8.

[0286] Study results

[0287] Tables 26-28 and Figure 10Show the effects of GDC-0570 and cobimetinib on tumor growth in a KRAS G12V mutant colorectal PDX model.

[0288] The measurement results of tumor volume on day 27 showed that the 2.5 mg / kg QD cobimetinib group had a 4% tumor growth inhibition (TGI) and a 97% T / C ratio (%). The 300 mg / kg QD GDC-0570 group had a 52% tumor growth inhibition (TGI) and a 63% T / C ratio (%). The combination of 300 mg / kg of GDC-0570 plus 2.5 mg / kg of cobimetinib had a >100% tumor growth inhibition (TGI) and a 26% T / C ratio (%).

[0289] Compared with the vehicle group, only the combination group showed significant tumor growth inhibition. Neither of the single-therapy groups showed significant tumor inhibition.

[0290] Compared with GDC-0570 or cobimetinib monotherapy, the combination therapy of GDC-0570 and cobimetinib showed superior efficacy.

[0291] Table 26: Effects of GDC-0570 combination on tumor volume in CRC PDX model CRC051

[0292]

[0293] “ / ” indicates not applicable

[0294] Table 27: Summary of efficacy relative to vehicle in PDX model CRC051

[0295]

[0296] Table 28: Summary of efficacy among treatment groups in PDX model CRC051

[0297]

[0298] P < 0.05 means statistically significant.

[0299] Although the foregoing has been described in considerable detail for purposes of clear understanding by way of illustration and example, such description and examples should not be construed as limiting the scope of the disclosure. All patents and scientific literature cited herein are hereby expressly incorporated by reference in their entirety.

Claims

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

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

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

6. The method according to any one of claims 1-5, wherein the KRAS inhibitor is selected from 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 inhibitor, pan-KRAS inhibitor, KRAS-SOS1 interaction inhibitor or KRAS signaling inhibitor.

7. The method according to claim 6, wherein the KRAS G12C inhibitor is selected from sotorasib (AMG 510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI 1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, and pharmaceutically acceptable salts thereof; wherein the KRAS G12D inhibitor is selected from MRTX1133, JAB-22000, RMC-9805 (RM-036), and pharmaceutically acceptable salts thereof; wherein the KRAS G13C inhibitor is selected from RMC-8839 and pharmaceutically acceptable salts thereof; wherein the KRAS G12V inhibitor is selected from JAB-23000 and pharmaceutically acceptable salts thereof; wherein the pan-KRAS inhibitor is selected from RMC-6236, BBP-454, and pharmaceutically acceptable salts thereof; wherein the KRAS-SOS1 interaction inhibitor is selected from BI 1701963, BAY-293, RMC-5845, BI-3406, SDGR5, and pharmaceutically acceptable salts thereof; wherein the KRAS signaling inhibitor is selected from cobimetinib and pharmaceutically acceptable salts thereof.

8. The method according to claim 6, wherein the PIM kinase inhibitor is selected from GDC-0570, GNE-1571, GNE-5775, GDC-0339 and GNE-5652, and pharmaceutically acceptable salts thereof; and the KRAS G12C inhibitor is selected from sotorasib (AMG 510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI 1823911, 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 1-8, 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. The method of any one of claims 1-9, wherein the cancer has a KRAS p.G12C, G12V, G12D, G13C, G13D, Q61H, Q61L, Q61R, K117N mutation or a combination thereof.

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

12. The method of any one of claims 1-11, wherein the cancer is selected from lung cancer, colorectal cancer, pancreatic cancer, appendiceal 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 cancer, cervical cancer, gastroenteropancreatic neuroendocrine tumor, endometrioid carcinoma, germ cell tumor, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord-stromal tumor, intrahepatic cholangiocarcinoma, histiocytosis, anal cancer, melanoma, mature B-cell neoplasm, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland adenocarcinoma, breast cancer, renal cell carcinoma, and bone cancer.

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

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

15. The method according to claim 13, wherein the acquired KRAS inhibitor resistance is selected from resistance to sotorasib (AMG 510), ARS-853, ARS-1620, ARS-3248 (JNJ-74699157), adagrasib (MRTX849), MRTX1257, LY3499446, LY3537982, BI 1823911, RG6330 (GDC-6036), RMC-6291, RMC-6236, AZD4625, D-1553, JDQ443, MK-1084, MRTX1133, JAB-22000, RMC-9805 (RM-036), RMC-8839, JAB-23000, cobimetinib, RMC-6236, BBP-454, BI 1701963, 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 as described in any one of claims 2-5, and the KRAS inhibitor is as described in any one of claims 6-8.

17. The combination according to claim 16, wherein the combination is used for treating cancer as described in any one of claims 10-12.

18. Use of a combination of a PIM kinase inhibitor and a KRAS inhibitor in the preparation of a medicament for treating cancer, wherein the PIM kinase inhibitor is as described in any one of claims 2-5, and the KRAS inhibitor is as described in any one of claims 6-8.

19. The use according to claim 18, wherein the cancer is as described in any one of claims 10-15.

20. A pharmaceutical composition comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable excipient, wherein the PIM kinase inhibitor is as described in any one of claims 2-5, and the KRAS inhibitor is as described in any one of claims 6-8.

21. The pharmaceutical composition according to claim 20, wherein the pharmaceutical composition is used for treating cancer as described in any one of claims 10-15.

22. A kit comprising a PIM kinase inhibitor, a KRAS inhibitor and a pharmaceutically acceptable excipient, wherein the PIM kinase inhibitor is as described in any one of claims 2-5, and the KRAS inhibitor is as described in any one of claims 6-8.

23. The kit according to claim 22, wherein the kit is used for treating cancer as described in any one of claims 10-15.

24. A method for treating cancer with KRAS mutations in human subjects in need thereof, the method 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 for use as a medicament for treating cancer with KRAS mutations; or the 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 preparation of a medicament for treating cancer with KRAS mutations.

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

26. The method or compound or use according to claim 24 or 25, wherein the cancer is resistant to KRAS inhibitors, and the KRAS inhibitor resistance includes intrinsic resistance, acquired resistance or adaptive resistance.

27. The method or compound or use according to claims 24-26, wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, pancreatic cancer, appendiceal 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 cancer, cervical cancer, gastroenteropancreatic neuroendocrine tumor, endometrioid cancer, germ cell tumor, esophagogastric cancer, bladder cancer, ovarian cancer, sex cord stromal tumor, intrahepatic cholangiocarcinoma, histiocytosis, anal cancer, melanoma, mature B-cell tumor, soft tissue sarcoma, gastrointestinal stromal tumor, head and neck cancer, glioma, prostate cancer, salivary gland cancer, breast cancer, renal cell cancer and bone cancer.

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