Pharmaceutical composition for preventing or treating cancer comprising SOS1 inhibitor and KRAS inhibitor

Abstract

The present invention relates to: a pharmaceutical composition for preventing or treating cancer, comprising an SOS1 inhibitor and a KRAS inhibitor; a combination for co-administration; a kit; and uses thereof, and can exhibit a remarkably excellent effect on the prevention and / or treatment of cancer compared to single administration.

Description

[0001]

Description of the Invention

[0002]

Title of Invention

[0003] Pharmaceutical composition for the prevention or treatment of cancer comprising S0S1 inhibitors and KRAS inhibitors

[0004]

Technology Field

[0005] The present invention relates to a combination therapy of S0S1 inhibitors and KRAS inhibitors useful for treating cancer. In particular, the present invention relates to pharmaceutical compositions, combinations, kits, and uses thereof comprising a compound (S0S1 inhibitor) that inhibits GTP-mediated nucleotide exchange of SOS1 (Son of seven less 1) and a KRAS (KRAS G12C, KRAS G12D, and / or KRAS G12V) inhibitor.

[0006] This research was conducted with funding from the Ministry of Science and ICT, the Ministry of Trade, Industry and Energy, and the Ministry of Health and Welfare, and with support from the National New Drug Development Project of the National New Drug Development Center (Project No.: RS-2023-00217674).

[0007]

Background Techniques

[0008] The Renin-Angiotensin System (RAS) family of proteins includes KRAS, NRAS, and HRAS, and their mutations exist in cells in either a Guanosine Triphosphate (GTP)-bound or Guanosine Diphosphate (GDP)-bound state. RAS family proteins inherently possess weak GTPase activity and slow nucleotide exchange rates. Binding of guanine nucleotide exchange factors (GEFs), such as Son of Seven Less 1 (SOSl), to RAS proteins promotes the release of GDP from the RAS proteins and enables GTP binding. When in the GTP-bound state, RAS family proteins become activated and interact with downstream effector proteins such as C-RAF and PI3K (Phosphatidyl Inositol-3-kinase). These pathways influence various cellular processes such as proliferation, survival, metabolism, motility, angiogenesis, immunity, and growth. Cancer-associated mutations in RAS family proteins inhibit GAP (GTPase-activating protein)-induced GTPase activity to increase GTP-bound active RAS family proteins. This leads to sustained activation of effector pathways (e.g., MEK / ERK, PI3K / AKT / mT0R, RalGDS pathways) downstream of RAS family proteins.

[0009] KRAS (Kirsten Rat Sarcoma 2 Viral Oncogene Homolog) is a small GTPase and one of the Ras family oncogenes. KRAS acts as a molecular switch that cycles between inactive (GDP-bound) and active (GTP-bound) states, and regulates various processes including cell proliferation by transmitting upstream cellular signals received from various tyrosine kinases to downstream pathways.

[0010] The role of activated KRAS in malignant tumors was observed 30 years ago. KRAS mutations (e.g., amino acids G12, G13, Q61, A146) are found in various human cancers, including lung, colorectal, and pancreatic cancers. Abnormal expression of KRAS accounts for up to 20% of all cancers. Oncogenic KRAS mutations that stabilize GTP binding and induce activation of KRAS and downstream signaling have been reported in 25–30% of lung adenocarcinomas. Single nucleotide substitutions resulting in missense mutations at codons 12 and 13 of the KRAS primary amino acid sequence account for approximately 40% of lung adenocarcinomas.

[0011] The role of KRAS in malignant tumors is well known, and with the discovery of frequent KRAS mutations (KRAS G12C, KRAS G12D, KRAS G12V) in various tumor types, KRAS has become a very promising target for anticancer therapy in the pharmaceutical industry. Despite research efforts to develop KRAS inhibitors as cancer treatments, no KRAS inhibitor has demonstrated sufficient efficacy to receive regulatory approval. Consequently, there is still a need to develop alternative approaches to maximize the efficacy and effectiveness of KRAS inhibitors as cancer treatments.

[0012] The SOS (Son of Seven less) protein exists in two isomorphic states, S0S1 and S0S2, but only S0S1 is phosphorylated by Doke. Growth factor-induced phosphorylation of S0S1 is largely mediated by Doke, which phosphorylates at least four serine residues in the C-terminal region of S0S1. This suggests that S0S1 plays an important role in the negative feedback regulation of the KRAS pathway.

[0013] S0S1 is the human homolog of the first discovered Drosophila protein, SOS. S0S1 has two sites that bind to RAS family proteins: a catalytic site that binds to GDP-binding RAS family proteins and an allosteric site that binds to GTP-binding RAS family proteins. KRAS proteins exchange GDP for GTP for activation, and S0S1, a GEF, activates KRAS by increasing the conversion to GTP.

[0014] Therefore, S0S1 activity regulates the active state of KRAS. Since S0S1 inhibitors significantly reduce pERK activity in cells of various KRAS mutants (KRAS G12C, KRAS G12D, KRAS G12V), S0S1 inhibitors exhibit pan-KRAS inhibitory activity regardless of the KRAS mutant type.

[0015] Accordingly, the inventors of the present invention completed the present invention by studying the combined administration of S0S1 inhibitors and KRAS inhibitors and confirming that the combination therapy described below has excellent efficacy as a preventive or therapeutic agent for cancer.

[0016] Prior art literature

[0017] Patent Literature Republic of Korea Published Patent No. 10-2025-0047111

[0018] Non-patent literature

[0019] Alamgeer et al. , Novel therapeutic targets in non-smal 1 cel 1 lung cancer , (2013) Current Op in Pharmcol . 13:394-401

[0020]

Description of the Invention

[0021]

Technical Challenges

[0022] The present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by the following chemical formula I or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered in combination with a KRAS inhibitor:

[0023] [Chemical formula U

[0024]

[0025] The present invention relates to (i) a first pharmaceutical composition comprising a compound represented by the formula I or a pharmaceutically acceptable salt thereof; and

[0026] (ii) Provides a combination of pharmaceutical compositions for co-administration for the prevention or treatment of cancer, comprising a second pharmaceutical composition comprising a KRAS inhibitor.

[0027] The present invention provides a kit for combination administration for the prevention or treatment of cancer comprising: (i) a first pharmaceutical composition comprising a compound represented by the formula I or a pharmaceutically acceptable salt thereof; and (ii) a second pharmaceutical composition comprising a KRAS inhibitor.

[0028] The present invention provides the use of the above pharmaceutical composition, combination, or kit for the prevention or treatment of cancer.

[0029] The present invention provides the use of the pharmaceutical composition, combination, or kit for the manufacture of a drug for the prevention or treatment of cancer.

[0030] The present invention provides a method for the prevention or treatment of cancer comprising the step of administering the above pharmaceutical composition, combination, or kit to an individual in need thereof.

[0031]

Technical Solution

[0032] The present invention will be described in more detail below. All combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the following detailed description.

[0033] Pharmaceutical composition for the prevention or treatment of cancer

[0034] The present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by the following formula I or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered in combination with a KRAS inhibitor:

[0035] [Chemical Formula I]

[0036]

[0037] In the present invention, the compound represented by the above formula I is named “N-((R)-1-(3-(difluoromethyl)-2-fluorphenyl)ethyl)-6-((S)-hexahydropyrazino[2,1-c][1,4]oxazine-8(1H)-yl)cinnoline-4-amine”.

[0038] In the present invention, “pharmaceutically acceptable” may mean physiologically acceptable and, when administered to an individual, not typically cause gastrointestinal disorders, allergic reactions such as dizziness, or similar reactions.

[0039] In the present invention, “pharmaceuticalally acceptable salt” refers to a salt commonly used in the pharmaceutical industry, and the pharmaceutically acceptable salt of the present invention may be prepared by conventional methods known to those skilled in the art. For example, the pharmaceutically acceptable salt may be an inorganic ionic salt prepared with calcium, potassium, sodium, or magnesium, etc.; an inorganic acid salt prepared with hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, or sulfuric acid, etc.; an organic acid salt prepared with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carboxylic acid, or vanillic acid, etc. Sulfonate salts prepared with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or naphthalenesulfonic acid; amino acid salts prepared with glycine, arginine, or lysine; and amine salts prepared with trimethylamine, trithylamine, ammonia, pyridine, or picolin, but the types of salts referred to in the present invention are not limited by these listed salts.

[0040] The pharmaceutically acceptable salt of the present invention can be prepared by conventional methods known to those skilled in the art.

[0041] In one embodiment, the pharmaceutically acceptable salt of the compound represented by the formula I may be a hydrochloric acid salt. Specifically, the hydrochloric acid salt may be a monohydrochloric acid salt, a 1.5-hydrochloric acid salt, or a dihydrochloric acid salt, and more specifically, the hydrochloric acid salt may be a dihydrochloric acid salt, but is not limited thereto.

[0042] Specifically, a pharmaceutically acceptable salt of the compound represented by Formula I above may be a compound represented by Formula II below, referred herein as an S0S1 inhibitor, compound A, and / or example compound:

[0043] [Chemical Formula II]

[0044]

[0045] N

[0046] According to one embodiment, the hydrochloride salt of the compound represented by Formula I of the present invention inhibits the binding, i.e., interaction, between KRAS mutant proteins (KRAS G12C mutant, KRAS G12D mutant, or KRAS G12V mutant) and S0S1 protein (see Experimental Example 1), and exhibits pan-KRAS inhibitory activity that inhibits the in vitro proliferation of cell lines having KRAS G12C mutant and KRAS G12D mutant (see Experimental Example 2). Furthermore, the hydrochloride salt of the compound represented by Formula I according to the present invention exhibits an excellent drug distribution profile compared to plasma in tumors of a KRAS G12C mutant pancreatic cancer xenograft model (see Experimental Example 3).

[0047] In one embodiment, the KRAS inhibitor may be a KRAS G12C inhibitor, a KRAS G12D inhibitor, or a KRAS G12V inhibitor.

[0048] In the present invention, “S0S1 inhibitor” and “KRAS inhibitor” refer to molecules capable of binding to S0S1 and KRAS, respectively, reducing the expression levels of S0S1 protein and KRAS protein, respectively, or neutralizing, blocking, inhibiting, reducing, or interfering with the biological activity of S0S1 and KRAS, respectively. Specifically, the “KRAS” includes a KRAS variant.

[0049] The above “activity” specifically includes the activity of S0S1 or KRAS, as well as biological activity mediated by S0S1 or KRAS. Specifically, “S0S1 inhibitors” and “KRAS inhibitors” each reduce or inhibit the expression levels of S0S1 protein and KRAS protein, or the biological activity mediated by S0S1 and KRAS, respectively, by at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more.

[0050] In the present invention, “inhibitor” may be used interchangeably with “inhibitory compound,” “inhibitor,” and “inhibitory compound.”

[0051] In the present invention, “KRAS G12C”, “KRAS G12D”, and “KRAS G12V” refer to variants in which the 12th residue glycine (Gly, G) of the KRAS protein is substituted with cysteine ​​(Cys, C), aspartic acid (Asp, D), and valine (Vai, V), respectively. “KRAS G12C”, “KRAS G12D”, and “KRAS G12V” may be used interchangeably with “KRAS(G12C)”, “KRAS(G12D)”, and “KRAS(G12V)”, respectively. In one embodiment, the KRAS G12C inhibitor may be one or more selected from the group consisting of sotorasib, adagrasib, olomorasib, divarasib, glecirasib, calderasib, JNJ-74699157, ZG-19018, YL-15293, ICP-915, BI-1823911, BEBT-607, ERAS-3490, and BPI-421286. Specifically, the KRAS G12C inhibitor may be sotorasib or adagrasib.

[0052] In one embodiment, the KRAS G12D inhibitor may be one or more selected from the group consisting of MRTX1133, Zoldonrasib, HRS-4642, INCB161734, QTX3034, AZD0022, and BI-KRASG12D3. Specifically, the KRAS G12D inhibitor may be MRTX1133.

[0053] In one embodiment, a pharmaceutical composition comprising a compound represented by the formula I or a pharmaceutically acceptable salt thereof; and a KRAS inhibitor may each be formulated and administered simultaneously or sequentially.

[0054] In one embodiment, a pharmaceutical composition comprising a compound represented by the formula I or a pharmaceutically acceptable salt thereof; and a KRAS inhibitor may each be included in the form of a separate formulation.

[0055] In one embodiment, a pharmaceutical composition comprising a compound represented by the formula I or a pharmaceutically acceptable salt thereof; and a KRAS inhibitor may be included in a mixed form.

[0056] For example, the compound represented by the above formula I or its pharmaceutically acceptable salt and a KRAS inhibitor may each be independently formulated into separate unit dosage forms (e.g., tablets, capsules, etc.) and administered in combination. Additionally, the compound represented by the above formula I or its pharmaceutically acceptable salt and a KRAS inhibitor may be formulated into a complex formulation and administered simultaneously.

[0057] In the present invention, “administration” means introducing a specific substance to an individual by an appropriate method.

[0058] In the present invention, “individual” refers to all animals, including rats, mice, and livestock, including humans, that have developed or may develop a disease; specifically, it may be mammals including humans, but is not limited thereto.

[0059] The pharmaceutical composition according to the present invention comprises a compound represented by the formula I or a pharmaceutically acceptable salt thereof for administration; and may further comprise one or more pharmaceutically acceptable carriers in addition to the KRAS inhibitor.

[0060] Pharmaceutically acceptable carriers are those commonly used in the art, and specifically may be lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidine, cellulose, water, syrup, methylcellulose, methyl hydroxybenzone, propyl hydroxybenzone, talc, magnesium stearate, or mineral oil, but are not limited thereto. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, a dispersant, a stabilizer, etc. In addition, the pharmaceutical composition of the present invention may be formulated using pharmaceutically acceptable carriers and excipients into oral formulations such as tablets, powders, granules, pills, capsules, suspensions, emulsions, liquids, emulsions, syrups, etc., external formulations, suppositories, or sterile injectable solutions, and may be manufactured in a unit dose form or contained in a multi-dose container. The formulation may be manufactured by conventional methods used in the art for formulation or by the methods disclosed in Remington's Pharmaceutical Science (19th ed., 1995), and may be formulated into various formulations depending on each disease or component.

[0061] The pharmaceutical composition of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the intended method, and the dosage may vary depending on the patient's body weight, age, gender, health status, diet, time of administration, method of administration, excretion rate, and severity of the disease. The compound of the present invention, or its pharmaceutically acceptable salt and KRAS inhibitor may be administered once or several times a day, but is not necessarily limited thereto.

[0062] In the present invention, “concurrent administration” does not mean only simultaneous administration, but refers to any form of administration in which the compound of the present invention, its pharmaceutically acceptable salt, and the KRAS inhibitor act together on an individual even when administered at different times, so that each substance can perform a level equivalent to or greater than its inherent function.

[0063] In the present invention, “prevention” means any act of suppressing cancer in an individual or delaying the onset of cancer by administering a compound according to the present invention or a pharmaceutically acceptable salt thereof and a KRAS inhibitor in combination.

[0064] In the present invention, “treatment” means any act in which the symptoms of cancer in an individual are improved or beneficially altered by the combined administration of a compound according to the present invention, a pharmaceutically acceptable salt thereof, and a KRAS inhibitor.

[0065] In one embodiment, the cancer may include a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation. In one embodiment, the cancer is a cancer including a KRAS G12C mutation, and the KRAS inhibitor may be a KRAS G12C inhibitor.

[0066] In one embodiment, the cancer is a cancer containing a KRAS G12D mutation, and the KRAS inhibitor may be a KRAS G12D inhibitor.

[0067] In one embodiment, the cancer is a cancer containing a KRAS G12V mutation, and the KRAS inhibitor may be a KRAS G12V inhibitor.

[0068] In one embodiment, the cancer may be one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma, and solid tumors. Specifically, the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, or small intestine cancer.

[0069] In one embodiment, the cancer may be one or more selected from the group consisting of pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, colorectal cancer containing a KRAS G12C mutation, pancreatic cancer containing a KRAS G12D mutation, non-small cell lung cancer containing a KRAS G12D mutation, colorectal cancer containing a KRAS G12D mutation, pancreatic cancer containing a KRAS G12V mutation, non-small cell lung cancer containing a KRAS G12V mutation, and colorectal cancer containing a KRAS G12V mutation. Specifically, the cancer may be pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, or colorectal cancer containing a KRAS G12C mutation. In one embodiment, the KRAS inhibitor is a KRAS G12C inhibitor, and the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, or small intestine cancer. Specifically, the KRAS inhibitor is a KRAS G12C inhibitor, and the cancer may be pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, or colorectal cancer containing a KRAS G12C mutation. More specifically, the KRAS inhibitor is sotorasib or adagrasib, and the cancer may be pancreatic cancer with a KRAS G12C mutation, non-small cell lung cancer with a KRAS G12C mutation, or colorectal cancer with a KRAS G12C mutation.More specifically, if the KRAS inhibitor is Sotorasib or Adagrasib, the non-small cell lung cancer or colorectal cancer containing the KRAS G12C mutation may be advanced or metastatic non-small cell lung cancer; or advanced or metastatic colorectal cancer.

[0070] In one embodiment, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal adenocarcinoma, appendiceal cancer, or small intestine cancer. Specifically, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer may be pancreatic cancer containing a KRAS G12D mutation, non-small cell lung cancer containing a KRAS G12D mutation, or colorectal cancer containing a KRAS G12D mutation. More specifically, the KRAS inhibitor is MRTX1133, and the cancer may be pancreatic cancer containing a KRAS G12D mutation. In one embodiment, the KRAS inhibitor is a KRAS G12V inhibitor, and the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, or small intestine cancer. Specifically, the KRAS inhibitor is a KRAS G12V inhibitor, and the cancer may be pancreatic cancer containing a KRAS G12V mutation, non-small cell lung cancer containing a KRAS G12V mutation, or colorectal cancer containing a KRAS G12V mutation.

[0071] According to one embodiment, the compound of the present invention or its pharmaceutically acceptable salt and a KRAS inhibitor are combined and treated with KRAS G12C mutant cancer cell lines (pancreatic cancer, non-small cell lung cancer and colorectal cancer) and KRAS G12D mutant cancer cell lines (pancreatic cancer) to exhibit a synergistic effect on inhibiting cancer cell growth (see Experimental Examples 4 and 5).

[0072] According to one embodiment, the combined administration of the compound of the present invention or its pharmaceutically acceptable salt and a KRAS inhibitor exhibits superior inhibition of tumor growth compared to the single administration of each of the compound of the present invention and the KRAS inhibitor in both KRAS G12C mutant xenograft mouse models (pancreatic cancer, non-small cell lung cancer, colorectal cancer) and KRAS G12D mutant xenograft mouse models (pancreatic cancer), and shows a synergistic effect (see Experimental Examples 6 and 7).

[0073] In addition, the compound of the present invention or a pharmaceutically acceptable salt thereof acts as an S0S1 inhibitor and effectively inhibits the interaction between KRAS mutant proteins and S0S1 proteins, and exhibits pan-KRAS inhibitory activity that inhibits cell growth and proliferation of various KRAS mutant cell lines.

[0074] Accordingly, the compound of the present invention or a pharmaceutically acceptable salt thereof can be administered in combination with a KRAS inhibitor to exhibit excellent preventive or therapeutic effects against cancers with KRAS activity (e.g., KRAS G12C mutant cancer, KRAS G12D mutant cancer, or KRAS G12V mutant cancer). In particular, the present invention has confirmed unexpected synergistic effects resulting from the aforementioned combination administration.

[0075] In one embodiment, the combined administration of a pharmaceutical composition comprising a compound represented by Formula I or a pharmaceutically acceptable salt thereof and a KRAS inhibitor may exhibit a synergistic effect in the prevention or treatment of cancer compared to the single administration of each of the pharmaceutical composition and the KRAS inhibitor.

[0076] The pharmaceutical composition of the present invention can be administered in combination with a KRAS inhibitor to inhibit the binding between KRAS and S0S1 proteins.

[0077] The pharmaceutical composition of the present invention can be administered in combination with a KRAS inhibitor to inhibit the growth, proliferation, or growth of KRAS-mutated cancer cells.

[0078] The pharmaceutical composition of the present invention can be administered in combination with a KRAS inhibitor to inhibit tumor growth of KRAS-mutated cancer.

[0079] The pharmaceutical composition of the present invention can be administered in combination with a KRAS inhibitor to exhibit significantly superior preventive and / or therapeutic effects against KRAS-mutated cancer.

[0080] Combinations and kits for combined administration for the prevention or treatment of cancer

[0081] The present invention comprises ( i) a first pharmaceutical composition comprising a compound represented by the following formula I or a pharmaceutically acceptable salt thereof:

[0082] [Chemical Formula I]

[0083]

[0084] (ii) Provides a combination of pharmaceutical compositions for co-administration for the prevention or treatment of cancer, comprising a second pharmaceutical composition comprising a KRAS inhibitor.

[0085] The present invention comprises ( i) a first pharmaceutical composition comprising a compound represented by the following formula I or a pharmaceutically acceptable salt thereof:

[0086] [Chemical Formula I]

[0087]

[0088] (ii) Provides a kit for combination administration for the prevention or treatment of cancer comprising a second pharmaceutical composition comprising a KRAS inhibitor.

[0089] The above “low release,” “KRAS G12C,” “KRAS G12D,” “KRAS G12V,” “prevention,” “treatment,” “concurrent administration,” etc. may be within the aforementioned range.

[0090] In one embodiment, the pharmaceutically acceptable salt of the compound represented by Formula I may be a hydrochloric acid salt. Specifically, the hydrochloric acid salt may be a monohydrochloric acid salt, a 1.5-hydrochloric acid salt, or a dihydrochloric acid salt, and more specifically, the hydrochloric acid salt may be a dihydrochloric acid salt, but is not limited thereto. Specifically, the pharmaceutically acceptable salt of the compound represented by Formula I may be a compound represented by the following Formula II, which is referred to herein as an S0S1 inhibitor, compound A, and / or example compound:

[0091] [Chemical Formula II]

[0092] 2HCI

[0093] H

[0094]

[0095] In one embodiment, the KRAS inhibitor may be a KRAS G12C inhibitor, a KRAS G12D inhibitor, or a KRAS G12V inhibitor.

[0096] In one embodiment, the KRAS G12C inhibitor may be one or more selected from the group consisting of sotorasib, adagrasib, olomorasib, divarasib, glecirasib, calderasib, JNJ-74699157, ZG-19018, YL-15293, ICP-915, BI-1823911, BEBT-607, ERAS-3490, and BPI-421286. Specifically, the KRAS G12C inhibitor may be sotorasib or adagrasib.

[0097] In one embodiment, the KRAS G12D inhibitor may be one or more selected from the group consisting of MRTX1133, Zoldonrasib, HRS-4642, INCB161734, QTX3034, AZD0022, and BI-KRASG12D3. Specifically, the KRAS G12D inhibitor may be MRTX1133.

[0098] In one embodiment, the first pharmaceutical composition and the second pharmaceutical composition may be administered together. In one embodiment, the first pharmaceutical composition and the second pharmaceutical composition may each be formulated and administered simultaneously or sequentially.

[0099] In one embodiment, the first pharmaceutical composition and the second pharmaceutical composition may each be included in the form of separate formulations.

[0100] In one embodiment, the first pharmaceutical composition and the second pharmaceutical composition may be included in a mixed form.

[0101] For example, the first pharmaceutical composition and the second pharmaceutical composition may each be independently formulated into separate unit dosage forms (e.g., tablets, capsules, etc.) and administered together. Additionally, the first pharmaceutical composition and the second pharmaceutical composition may be formulated into a combined formulation and administered simultaneously.

[0102] In one embodiment, the cancer may include a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation.

[0103] In one embodiment, the cancer is a cancer containing a KRAS G12C mutation, and the KRAS inhibitor may be a KRAS G12C inhibitor.

[0104] In one embodiment, the cancer is a cancer containing a KRAS G12D mutation, and the KRAS inhibitor may be a KRAS G12D inhibitor.

[0105] In one embodiment, the cancer is a cancer containing a KRAS G12V mutation, and the KRAS inhibitor may be a KRAS G12V inhibitor.

[0106] In one embodiment, the cancer may be one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma, and solid tumors. Specifically, the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, or small intestine cancer.

[0107] In one embodiment, the cancer may be one or more selected from the group consisting of pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, colorectal cancer containing a KRAS G12C mutation, pancreatic cancer containing a KRAS G12D mutation, non-small cell lung cancer containing a KRAS G12D mutation, colorectal cancer containing a KRAS G12D mutation, pancreatic cancer containing a KRAS G12V mutation, non-small cell lung cancer containing a KRAS G12V mutation, and colorectal cancer containing a KRAS G12V mutation. Specifically, the cancer may be pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, or colorectal cancer containing a KRAS G12C mutation.

[0108] In one embodiment, the KRAS inhibitor is a KRAS G12C inhibitor, and the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, or small intestine cancer. Specifically, the KRAS inhibitor is a KRAS G12C inhibitor, and the cancer may be pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, or colorectal cancer containing a KRAS G12C mutation. More specifically, the KRAS inhibitor is sotorasib or adagrasib, and the cancer may be pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, or colorectal cancer containing a KRAS G12C mutation. More specifically, if the KRAS inhibitor is sotorasib or adagrasib, the non-small cell lung cancer or colorectal cancer containing a KRAS G12C mutation may be advanced or metastatic non-small cell lung cancer; or advanced or metastatic colorectal cancer.

[0109] In one embodiment, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal adenocarcinoma, appendiceal cancer, or small intestine cancer. Specifically, the KRAS inhibitor is a KRAS G12D inhibitor, and the cancer may be pancreatic cancer containing a KRAS G12D mutation, non-small cell lung cancer containing a KRAS G12D mutation, or colorectal cancer containing a KRAS G12D mutation. More specifically, the KRAS inhibitor is MRTX1133, and the cancer may be pancreatic cancer containing a KRAS G12D mutation.

[0110] In one embodiment, the KRAS inhibitor is a KRAS G12V inhibitor, and the cancer may be lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, or small intestine cancer. Specifically, the KRAS inhibitor is a KRAS G12V inhibitor, and the cancer may be pancreatic cancer containing a KRAS G12V mutation, non-small cell lung cancer containing a KRAS G12V mutation, or colorectal cancer containing a KRAS G12V mutation.

[0111] The compound of the present invention or a pharmaceutically acceptable salt thereof acts as an S0S1 inhibitor, effectively inhibiting the interaction between KRAS mutant proteins and S0S1 proteins, and exhibits pan-KRAS inhibitory activity that inhibits cell growth and proliferation of various KRAS mutant cell lines. Therefore, the compound of the present invention or a pharmaceutically acceptable salt thereof, when co-administered with a KRAS inhibitor, can exhibit excellent preventive or therapeutic effects against cancers with KRAS activity (e.g., KRAS G12C mutant cancer, KRAS G12D mutant cancer, or KRAS G12V mutant cancer). In particular, the present invention has confirmed unexpected synergistic effects resulting from the above co-administration.

[0112] In one embodiment, the combined administration of the first pharmaceutical composition and the second pharmaceutical composition may exhibit a synergistic effect in the prevention or treatment of cancer compared to the single administration of each of the first pharmaceutical composition and the second pharmaceutical composition.

[0113] The combination and kit for co-administration of the present invention can inhibit the binding between KRAS and S0S1 proteins.

[0114] The combination and kit for combined administration of the present invention can inhibit the growth, proliferation, or growth of KRAS mutant cancer cells.

[0115] The combination and kit for combined administration of the present invention can inhibit tumor growth of KRAS-mutated cancer.

[0116] The combination of substances and kits for combined administration of the present invention can exhibit significantly superior preventive and / or therapeutic effects against KRAS-mutated cancers.

[0117] Treatment methods and uses

[0118] The present invention provides the use of the above pharmaceutical composition, combination, or kit for the prevention or treatment of cancer.

[0119] The present invention provides a use of the pharmaceutical composition, combination, or kit for manufacturing a drug for the prevention or treatment of cancer. The present invention provides a method for the prevention or treatment of cancer comprising the step of administering the pharmaceutical composition, combination, or kit to an individual in need thereof.

[0120] In one embodiment, the administering step may be administering the pharmaceutical composition, combination, or kit to an individual requiring it in a therapeutically effective amount.

[0121] In the present invention, the term “therapeutic effective dose” refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment and that does not cause side effects. This amount may be determined by a person skilled in the art based on factors including the patient’s gender, age, weight, health status, type and severity of the disease, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration, release rate, duration of treatment, drugs used in combination or concurrently, and other factors well known in the medical field. It is desirable to apply a specific therapeutic effective dose for a specific patient differently based on various factors including the specific composition, the type and degree of response to be achieved, whether different preparations are used in some cases, the patient’s age, weight, general health status, gender and diet, time of administration, route of administration and release rate of the composition, duration of treatment, drugs used with or concurrently with the specific composition, and similar factors well known in the pharmaceutical field. The above “low release,” “KRAS G12C,” “KRAS G12D,” “KRAS G12V,” “prevention,” “treatment,” “concurrent administration,” etc. may be within the aforementioned range.

[0122] In addition, the present invention comprises the following items (1) to (48) relating to pharmaceutical compositions, combinations, kits and uses thereof. (1) The present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by the following chemical formula I or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered in combination with a KRAS inhibitor:

[0123] [Chemical Formula I]

[0124]

[0125] (2) In the above (1), the pharmaceutically acceptable salt of the compound represented by chemical formula I may be a hydrochloric acid salt.

[0126] (3) In the above (1) or (2), the hydrochloric acid salt may be a monohydrochloric acid salt, a 1.5-hydrochloric acid salt, or a dihydrochloric acid salt.

[0127] (4) In any one of (1) to (3) above, the KRAS inhibitor may be a KRAS G12C inhibitor, a KRAS G12D inhibitor, or a KRAS G12V inhibitor.

[0128] (5) In any one of (1) to (4) above, the KRAS G12C inhibitor may be one or more selected from the group consisting of Sotorasib, Adagrasib, Olomorasib, Divarasib, Glecirasib, Calderasib, JNJ-74699157, ZG-19018, YL-15293, I CP-915, BI-1823911, BEBT-607, ERAS-3490 and BPI-421286. (6) In any one of (1) to (4) above, the KRAS G12D inhibitor may be one or more selected from the group consisting of MRTX1133, Zoldonrasib, HRS-4642, INCB161734, QTX3034, AZD0022 and BI-KRASG12D3.

[0129] (7) A pharmaceutical composition comprising a compound represented by chemical formula I or a pharmaceutically acceptable salt thereof in any one of (1) to (6) above; and a KRAS inhibitor may each be formulated and administered simultaneously or sequentially.

[0130] (8) A pharmaceutical composition comprising a compound represented by chemical formula I or a pharmaceutically acceptable salt thereof in any one of (1) to (6) above; and a KRAS inhibitor may each be included in the form of a separate formulation or a mixed form.

[0131] (9) In any one of (1) to (8) above, the cancer may include a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation.

[0132] (10) In any one of (1) to (5) and (7) to (9) above, the cancer is a cancer containing a KRAS G12C mutation, and the KRAS inhibitor may be a KRAS G12C inhibitor.

[0133] (11) In any one of (1) to (4) and (6) to (9) above, the cancer is a cancer containing a KRAS G12D mutation, and the KRAS inhibitor may be a KRAS G12D inhibitor.

[0134] (12) In any one of (1) to (11) above, the cancer may be one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma and solid tumors. (13) In any one of (1) to (12) above, the cancer may be one or more selected from the group consisting of pancreatic cancer with a KRAS G12C mutation, non-small cell lung cancer with a KRAS G12C mutation, colorectal cancer with a KRAS G12C mutation, pancreatic cancer with a KRAS G12D mutation, non-small cell lung cancer with a KRAS G12D mutation, colorectal cancer with a KRAS G12D mutation, pancreatic cancer with a KRAS G12V mutation, non-small cell lung cancer with a KRAS G12V mutation, and colorectal cancer with a KRAS G12V mutation.

[0135] (14) The present invention comprises a first pharmaceutical composition comprising ( i ) a compound represented by the following formula I or a pharmaceutically acceptable salt thereof:

[0136] [Chemical Formula I]

[0137]

[0138] (ii) Provides a combination of pharmaceutical compositions for co-administration for the prevention or treatment of cancer, comprising a second pharmaceutical composition comprising a KRAS inhibitor.

[0139] (15) In the above (14), the pharmaceutically acceptable salt of the compound represented by chemical formula I may be a hydrochloric acid salt.

[0140] (16) In the above (14) or (15), the hydrochloric acid salt may be a monohydrochloric acid salt, a 1.5-hydrochloric acid salt, or a dihydrochloric acid salt. (17) In any one of the above (14) to (16), the KRAS inhibitor may be a KRAS G12C inhibitor, a KRAS G12D inhibitor, or a KRAS G12V inhibitor.

[0141] (18) In any one of (14) to (17) above, the KRAS G12C inhibitor may be one or more selected from the group consisting of Sotorasib, Adagrasib, Olomorasib, Divarasib, Glecirasib, Calderasib, JNJ-74699157, ZG-19018, YL-15293, I CP-915, BI-1823911, BEBT-607, ERAS-3490 and BPI-421286.

[0142] (19) In any one of (14) to (17) above, the KRAS G12D inhibitor may be one or more selected from the group consisting of MRTX1133, Zoldonrasib, HRS-4642, INCB161734, QTX3034, AZD0022 and BI-KRASG12D3.

[0143] (20) In any one of (14) to (19) above, the first pharmaceutical composition and the second pharmaceutical composition may each be formulated and administered simultaneously or sequentially.

[0144] (21) In any one of (14) to (19) above, the first pharmaceutical composition and the second pharmaceutical composition may each be included in the form of a separate formulation or a mixed form.

[0145] (22) In any one of (14) to (21) above, the cancer may include a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation.

[0146] (23) In any one of (14) to (18) and (20) to (22) above, the cancer is a cancer containing a KRAS G12C mutation, and the KRAS inhibitor may be a KRAS G12C inhibitor. (24) In any one of (14) to (17) and (19) to (22) above, the cancer is a cancer containing a KRAS G12D mutation, and the KRAS inhibitor may be a KRAS G12D inhibitor.

[0147] (25) In any one of the above (14) to (24), the cancer may be one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma and solid tumors.

[0148] (26) In any one of (14) to (25) above, the cancer may be one or more selected from the group consisting of pancreatic cancer with a KRAS G12C mutation, non-small cell lung cancer with a KRAS G12C mutation, colorectal cancer with a KRAS G12C mutation, pancreatic cancer with a KRAS G12D mutation, non-small cell lung cancer with a KRAS G12D mutation, colorectal cancer with a KRAS G12D mutation, pancreatic cancer with a KRAS G12V mutation, non-small cell lung cancer with a KRAS G12V mutation, and colorectal cancer with a KRAS G12V mutation.

[0149] (27) The present invention comprises a first pharmaceutical composition comprising a compound represented by the following formula I or a pharmaceutically acceptable salt thereof:

[0150] [Chemical Formula I]

[0151]

[0152] (ii) Provides a kit for combination administration for the prevention or treatment of cancer comprising a second pharmaceutical composition comprising a KRAS inhibitor.

[0153] (28) In the above (27), the pharmaceutically acceptable salt of the compound represented by chemical formula I may be a hydrochloric acid salt.

[0154] (29) In the above (27) or (28), the hydrochloric acid salt may be a monohydrochloric acid salt, a 1.5-hydrochloric acid salt, or a dihydrochloric acid salt.

[0155] (30) In any one of (27) to (29) above, the KRAS inhibitor may be a KRAS G12C inhibitor, a KRAS G12D inhibitor, or a KRAS G12V inhibitor.

[0156] (31) In any one of (27) to (30) above, the KRAS G12C inhibitor may be one or more selected from the group consisting of Sotoras ib, Adagras ib, Olomoras ib, Divaras ib, Gleciras ib, Calderas ib, JNJ-74699157, ZG-19018, YL-15293, I CP-915, BI-1823911, BEBT-607, ERAS-3490 and BPI-421286.

[0157] (32) In any one of (27) to (30) above, the KRAS G12D inhibitor may be one or more selected from the group consisting of MRTX1133, zodonrasib (Zo ldonrasib), HRS-4642, INCB161734, QTX3034, AZD0022 and BI-KRASG12D3. (33) In any one of (27) to (32) above, the first pharmaceutical composition and the second pharmaceutical composition may each be formulated and administered simultaneously or sequentially.

[0158] (34) In any one of (27) to (32) above, the first pharmaceutical composition and the second pharmaceutical composition may each be included in the form of a separate formulation or a mixed form.

[0159] (35) In any one of (27) to (34) above, the cancer may include a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation.

[0160] (36) In any one of the above (27) to (31) and (33) to (35), the cancer is a cancer containing a KRAS G12C mutation, and the KRAS inhibitor may be a KRAS G12C inhibitor.

[0161] (37) In any one of the above (27) to (30) and (32) to (35), the cancer is a cancer containing a KRAS G12D mutation, and the KRAS inhibitor may be a KRAS G12D inhibitor.

[0162] (38) In any one of the above (27) to (37), the cancer may be one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma and solid tumors.

[0163] (39) In any one of (27) to (38) above, the cancer may be one or more selected from the group consisting of pancreatic cancer with a KRAS G12C mutation, non-small cell lung cancer with a KRAS G12C mutation, colorectal cancer with a KRAS G12C mutation, pancreatic cancer with a KRAS G12D mutation, non-small cell lung cancer with a KRAS G12D mutation, colorectal cancer with a KRAS G12D mutation, pancreatic cancer with a KRAS G12V mutation, non-small cell lung cancer with a KRAS G12V mutation, and colorectal cancer with a KRAS G12V mutation.

[0164] (40) The present invention provides a pharmaceutical composition according to any one of (1) to (13) for the prevention or treatment of cancer.

[0165] (41) The present invention provides a combination of (14) to (26) according to any one of the above for the prevention or treatment of cancer.

[0166] (42) The present invention provides a kit according to any one of (27) to (39) for the prevention or treatment of cancer.

[0167] (43) The present invention provides a use of a pharmaceutical composition according to any one of (1) to (13) for the manufacture of a drug for the prevention or treatment of cancer.

[0168] (44) The present invention provides a use of a combination according to any one of (14) to (26) for the manufacture of a drug for the prevention or treatment of cancer.

[0169] (45) The present invention provides a use of a kit according to any one of (27) to (39) for the manufacture of a drug for the prevention or treatment of cancer.

[0170] (46) The present invention provides a method for preventing or treating cancer, comprising the step of administering a pharmaceutical composition according to any one of (1) to (13) to an individual who needs it.

[0171] (47) The present invention provides a method for preventing or treating cancer comprising the step of administering a combination according to any one of (14) to (26) to an individual in need thereof. (48) The present invention provides a method for preventing or treating cancer comprising the step of administering a kit according to any one of (27) to (39) to an individual in need thereof.

[0172]

Effects of the Invention

[0173] The present invention relates to a pharmaceutical composition for the prevention or treatment of cancer comprising an S0S1 inhibitor and a KRAS inhibitor, a combination for co-administration, a kit, and the use thereof, which can exhibit significantly superior effects in the prevention and / or treatment of cancer through a synergistic effect compared to the single administration of an S0S1 inhibitor and a KRAS inhibitor.

[0174]

Brief Description of the Drawing

[0175] Figure 1 shows the results of the 3D spheroid assay of S0S1 inhibitor compound A. IC5o (inhibition concentration 50) refers to the concentration that reduces cell growth by 50%.

[0176] Figure 2 shows the PK results of plasma and tumors in the group administered compound A alone.

[0177] Figure 3 shows the results of evaluating the cell growth inhibitory efficacy in KRASCG12C) mutant MIA PaCa-2 cell lines using the HSA synergy score when combined with a KRASCG12C) inhibitor and S0S1 inhibitor compound A.

[0178] Figure 4 shows the results of evaluating the cell growth inhibitory efficacy in the KRASCG12C) mutant NCI-H1373 cell line using the HSA synergy score when combined with a KRAS(G12C) inhibitor and S0S1 inhibitor compound A. Figure 5 shows the results of evaluating the cell growth inhibitory efficacy in the KRASCG12C) mutant SW837 cell line using the HSA synergy score when combined with a KRAS(G12C) inhibitor and S0S1 inhibitor compound A.

[0179] Figure 6 shows the results of evaluating the cell growth inhibitory efficacy in KRASCG12D) mutant AsPC-1 cell lines using the HSA synergy score when combined with a KRASCG12D) inhibitor and S0S1 inhibitor compound A.

[0180] Figure 7 shows the tumor growth curves in an MIA PaCa-2 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (9 mice per group; n=9). *** p < 0.001 indicates a significance level compared to the vehicle treatment group after one-way ANOVA analysis and post-hoc Tukey's test, and ### p < 0.01 indicates a significance level compared to the Adagrasib 10 mg / kg treatment group after unpaired two-way t-test. +++ p < 0.001 indicates a significant difference compared to the 25 mg / kg treatment group of Compound A, as determined by an independent sample t-test (unpaired two-way led T-test). QD (quaque die) means once a day.

[0181] Figure 8 shows the results of tumor weight measurements in an MIA PaCa-2 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered alone and in combination. Data are expressed as mean SEM (9 mice per group; n=9). *** p < 0.001 indicates a significance level compared to the vehicle-treated group after one-way ANOVA analysis and post-hoc Tukey's test, and ### p < 0.001 indicates a significance level compared to the Adagrasib 10 mg / kg treated group after unpaired two-way t-test. +++ p < 0.001 indicates a significant level compared to the 25 mg / kg compound A treatment group, based on an independent sample size, i.e., an unpaired two-way led T-test. QD (quaque die) means once a day.

[0182] Figure 9 shows the change in mouse body weight in an MIA PaCa-2 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed in mean SEM. QD (quake die) means once a day.

[0183] Figure 10 shows the tumor growth curves in an MIA PaCa-2 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered alone and in combination. Data are expressed as mean SEM (8-9 mice per group; n=8-9). *** p < 0.001 indicates a significance level compared to the vehicle-treated group after one-way ANOVA analysis and post-hoc Tukey's test, and ## p < 0.01 indicates a significance level compared to the Sotorasib 10 mg / kg-treated group after unpaired two-way t-test. +++ p < 0.001 indicates a significant difference compared to the 25 mg / kg treatment group of Compound A, as determined by an independent sample t-test (unpaired two-way led T-test). QD (quaque die) means once a day.

[0184] Figure 11 shows the results of tumor weight measurements in an MIA PaCa-2 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered alone and in combination. Data are expressed as mean SEM (8-9 mice per group; n=8-9). *** p < 0.001 indicates a significance level compared to the vehicle-treated group after one-way ANOVA analysis and post-hoc Tukey's test, and p < 0.05 indicates a significance level compared to the Sotorasib 10 mg / kg treated group after an unpaired two-way t-test. +++ p < 0.001 indicates a significant difference compared to the 25 mg / kg treatment group of Compound A, as determined by an independent sample t-test (unpaired two-way led T-test). QD (quaque die) means once a day.

[0185] Figure 12 shows the changes in mouse body weight in an MIA PaCa-2 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM. QD (quake die) means once a day.

[0186] Figure 13 shows tumor growth curves in an NCI-H1373 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). *** p < 0.01 indicates a significant level compared to the vehicle-treated group as determined by Tukey's multiple comparison test with one-way ANOVA analysis, and ### p < 0.001 indicates a significant level compared to the Adagrasib 10 mg / kg treatment group as determined by an unpaired two-way t-test. +++ p < 0.001 indicates a significant level compared to the compound A 50 mg / kg treatment group, based on an independent sample t-test (unpaired two-way led T-test). QD (quaque die) means once a day.

[0187] Figure 14 shows the results of tumor weight measurements in an NCI-H1373 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). * p < 0.05, *** p < 0.001 indicate the significance level compared to the vehicle-treated group as determined by Tukey's multiple comparison test along with one-way ANOVA analysis, and ### p < 0.001 indicates the significance level compared to the Adagrasib 10 mg / kg treatment group as determined by an unpaired two-way t-test. +++ p < 0.001 indicates a significant level compared to the compound A 50 mg / kg treatment group, as determined by an independent sample t-test (unpaired two-way led T-test). QD (quaque die) means once a day.

[0188] Figure 15 shows the change in mouse body weight in an NCI-H1373 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed in mean SEM. QD (quaque die) means once a day.

[0189] Figure 16 shows the tumor growth curves in an NCI-H1373 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (9-10 mice per group; n=9-10). ** p < 0.01, *** p < 0.01 indicates the significance level compared to the vehicle treatment group as determined by Tukey's multiple comparison test with one-way AN0VA analysis, and ### p < 0.001 indicates the significance level compared to the Sotorasib 30 mg / kg treatment group as determined by an unpaired two-way t-test. +++ p < 0.001 indicates a significant level compared to the 50 mg / kg treatment group of Compound A, based on an independent sample amount, i.e., an unpaired two-way led T-test. QD (quaque die) means once a day.

[0190] Figure 17 shows the results of tumor weight measurements in an NCI-H1373 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered alone and in combination. Data are expressed as mean SEM (9-10 mice per group; n=9-10). * p < 0.05, *** p < 0.01 indicates a significant level compared to the vehicle treatment group as determined by Tukey's multiple comparison test with one-way AN0VA analysis, and ### p < 0.001 indicates a significant level compared to the Sotorasib 30 mg / kg treatment group as determined by an unpaired two-way t-test. A p < 0.001 value indicates a significant level compared to the 50 mg / kg treatment group of Compound A, based on an independent sample amount, i.e., an unpaired two-way led T-test. QD (quake die) means once a day.

[0191] Figure 18 shows the change in mouse body weight in an NCI-H1373 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed in mean SEM. QD (quaque die) means once a day.

[0192] Figure 19 shows tumor growth curves in a SW837 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). A p < 0.001 indicates a significant level compared to the vehicle treatment group as determined by Tukey's multiple comparison test with one-way AN0VA analysis, and a p < 0.001 indicates a significant level compared to the Adagrasib 10 mg / kg treatment group as determined by an unpaired two-way t-test. A p < 0.001 value indicates the significance level compared to the 100 mg / kg compound A treatment group, as determined by an independent sample quantity t-test (unpaired two-way led T-test). QD (quaque die) means once a day.

[0193] Figure 20 shows the results of tumor weight measurements in a SW837 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). *** p < 0.001 indicates a significant level compared to the vehicle treatment group in Tukey's multiple comparison test with one-way AN0VA analysis, and ### p < 0.001 indicates a significant level compared to the Adagrasib 10 mg / kg treatment group in an unpaired two-way led T-test. A p < 0.001 value indicates a significant level compared to the 100 mg / kg treatment group of compound A, based on an independent sample amount, i.e., an unpaired two-way led T-test. QD (quaque die) means once a day.

[0194] Figure 21 shows the change in mouse body weight in a SW837 xenograft model when the KRASCG12C inhibitor Adagrasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed in mean SEM. QD (quaque die) means once a day.

[0195] Figure 22 shows tumor growth curves in a SW837 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). A p < 0.001 indicates a significant level compared to the vehicle treatment group as determined by Tukey's multiple comparison test with one-way AN0VA analysis, and a p < 0.001 indicates a significant level compared to the Sotorasib 30 mg / kg treatment group as determined by an unpaired two-way t-test. A p < 0.001 value indicates a significant level compared to the 100 mg / kg treatment group of Compound A, based on an independent sample quantity t-test (unpaired two-way led T-test). QD (quake die) means once a day.

[0196] Figure 23 shows the results of tumor weight measurements in a SW837 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). *** p < 0.01 indicates a significant level compared to the vehicle-treated group using Tukey's multiple comparison test with one-way ANOVA analysis, and ### p < 0.001 indicates a significant level compared to the Sotorasib 30 mg / kg treatment group using an unpaired two-way t-test. +++ p < 0.001 indicates a significant level compared to the compound A 100 mg / kg treatment group as determined by an unpaired two-sided t-test. QD (quaque die) means once a day.

[0197] Figure 24 shows the change in mouse body weight in a SW837 xenograft model when the KRASCG12C inhibitor Sotorasib and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed in mean SEM. QD (quaque die) means once a day.

[0198] Figure 25 shows tumor growth curves in an AsPC-1 xenograft model when a KRASCG12D inhibitor and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). *** p < 0.01 indicates a significance level compared to the vehicle treatment group as determined by post-hoc Tukey's test after one-way ANOVA analysis, ## p < 0.01 indicates a significance level compared to the MRTX1133 10 mg / kg treatment group as determined by an unpaired two-way t-test, and +++ p < 0.001 indicates a significance level compared to the compound A 50 mg / kg treatment group as determined by an unpaired two-way t-test. QD (quaque die) means once a day, and BID (bis in die) means twice a day.

[0199] Figure 26 shows the results of tumor weight measurements in an AsPC-1 xenograft model when a KRASCG12D inhibitor and the S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed as mean SEM (10 mice per group; n=10). *** p < 0.01 indicates a significance level compared to the vehicle treatment group as determined by post-hoc Tukey's test after one-way ANOVA analysis, # p < 0.05 indicates a significance level compared to the MRTX1133 10 mg / kg treatment group as determined by an unpaired two-tai led T-test, +++ p < 0.001 indicates a significance level compared to the compound A 50 mg / kg treatment group as determined by an unpaired two-tai led T-test. QD (quaque die) means once a day, and BID (bis in die) means twice a day.

[0200] Figure 27 shows the change in body weight of mice in an AsPC-1 xenograft model when a KRASCG12D inhibitor and S0S1 inhibitor compound A were administered as monotherapy and combination therapy. Data are expressed in mean SEM. QD (quaque die) means once a day, and BID (bis in die) means twice a day.

[0201]

Form for carrying out the invention

[0202] The present invention will be explained in more detail below through examples. These examples are intended to explain the invention more specifically, and the scope of the invention is not limited by these examples.

[0203] <Preparation Example> Synthesis of compound (N-((R)-l-(3-(difluoromethyl)-2-fluorphenyl)ethyl)~6~((S)-hexahydropyrazino[2, l~c][1, 4]oxazine-8(1H)-yl)cinnoline-4~amine dihydrochloride; Compound A)

[0204] (1) Synthesis of compound I (Chemical Formula I)

[0205]

[0206] 50 g of 6-bromo-4-chlorosinoline, 46.6 g of (R)-1-(3-(difluoromethyl)-2-fluorphenyl) ethanamine, and 107 mL of thylamine were added to diisopropyl and stirred at 140°C for 3 hours. The reaction was terminated by adding 1 L of an aqueous NH4Cl1 solution. The product was extracted using 1 L of methyl chloride (MC). Subsequently, the same extraction process was performed two more times. Afterward, g of MgS0410 was added to the MC layer, stirred, and filtered. After concentrating the organic layer, 33.4 g of (R)-6-bromo-N-(1-(3-(difluoromethyl)-2-fluorphenyl)ethyl)cinnoline-4-amine was obtained by column chromatography (yield 41.1%).

[0207] MS (ESI+) m / z 396, 398 (M+H) +

[0208] Subsequently, 3.2 g of the obtained (R)-6-bromo-N-(1-(3-(difluoromethyl)-2-fluorphenyl)ethyl)cinnoline-4-amine, 0.74 g of Pd2(dba)3, 0.77 g of xphos, and 24 mL of tetrahydrofuran (THF) were added and stirred. 1.38 g of (S)-octahydropyrazino [2,1-c][1,4]oxazine was diluted in 8 mL of THF and added, followed by the addition of 32 mL of Li HMDS (1.0 M in THF). The mixture was stirred at 80°C for 2 hours. The reaction was terminated using 200 mL of an aqueous solution of NH4Cl. The product was extracted using 200 mL of a mixed organic solvent (methyl chloride (MC): isopropyl alcohol (IPA) = 3:1). The same extraction process was then performed two more times. 40.6 g of MgS0 was added to the organic layer, stirred, and filtered. After concentrating the organic layer, column chromatography yielded 1.48 g of the desired compound, N-((R)-1-(3-(difluoromethyl)-2-fluorphenyl)ethyl)-6-((S)-hexahydropyrazino[2,1-c][1,4]oxazine-8(1H)-yl)cinnoline-4-amine (yield 40.1%).

[0209]

[0210] NMR (400 MHz, DMSO— d6) 5 8.10 (s, 1H) , 7.96 (d, J = 9.2 Hz, 1H) , 7.72 (dd, J = 2.4 Hz, 6.8 Hz, 1H) , 7.59 (t , J = 7.2 Hz, 1H) , 7.54 (t , J = 7.2 Hz, 1H) , 7.46 (d, J = 2.4 Hz, 1H) , 7.42 (s, 0.25H) , 7.34 (s, 0.25H) , 7.31 (d, J = 3.2 Hz, 1H) , 7.28 (d, J = 4.0 Hz, 1H) , 7.27 (s, 0.25H) , 7.14 (s, 0.25 H) , 5.25-5.18 (m, 1H) , 4.02 (d, J = 11.6 Hz, 1H) , 3.85-3.79 (m, 3H) , 3.61-3.55 (m, 1H) , 3.25 (t , J = 10.4 Hz , 2H) , 3.00-2.92 (m, 2H) , 2.74 (d, J = 11.2 Hz , 1H) , 2.41-2.24 (m, 3H) , 1.70 (d, J = 6.8 Hz , 3H)

[0211] MS (ESI+) m / z 458 (M+H) +

[0212] (2) 실시에 화합물 (화학식 I

[0213]

[0214] Dihydrochloride of compound; preparation of compound A) 200 mL of acetone and 10.0 g of N-((R)-1-(3-(difluoromethyl)-2-fluorphenyl)ethyl)-6-((S)-hexahydropyrazino [2,1-c][1,4]oxazine-8(1H)-yl)cinnoline-4-amine were added to a reactor, and the mixture was heated to 40–45°C. A solution of 4.13 mL of a 35–37% aqueous hydrochloric acid solution diluted with 200 mL of acetone was slowly added dropwise over 30 minutes at the same temperature, stirred for 1 hour, cooled to 20–25°C, and stirred for an additional 1 hour. Subsequently, the resulting solid was filtered and vacuum dried at 40 to 60 °C to obtain 9.51 g of N-((R)-1-(3-(difluoromethyl)-2-fluorphenyl)ethyl)-6-((S)-hexahydropyrazino[2,1-c][1,4]oxazine-8(1H)-yl)cinnoline-4-amine dihydrochloride anhydride (Form 1) (yield 82%).

[0215] <Experimental Example 1> Evaluation of the Interaction Inhibitory Ability of KRAS(G12C, G12D, G12V)-SOS1 and KRAS(G12C)-SOS2

[0216] To confirm the inhibitory ability of the S0S1 inhibitory compound according to the present invention (Example compound; Compound A) on the interaction between KRASCG12C, G12D, G12V) proteins and S0S1 protein, and the inhibitory ability on the interaction between KRAS(G12C) protein and S0S2 protein, it was evaluated using an HTRF-based KRASCG12C, G12D, G12V)-SOS1 and KRAS(G12C)-S0S2 interaction inhibitory efficacy evaluation method.

[0217] Specifically, to evaluate the inhibitory effect of the interaction between KRAS (G12C, G12D, G12V) proteins and S0S1 proteins, KRAS (G12C, G12D, G12V) proteins were reacted with an α-GST Tb antibody for 1 hour, then a 1.5-fold solution of S0S1 was prepared in a buffer and 10 M was dispensed into the reaction plate. Subsequently, GTP was added to the KRAS / α-GST Tb antibody. Afterward, the reaction was started by dispensing 5 M of the S0S1 mixture with the compound reacted for 15 minutes. The exchange reaction of GDP to GTP-DY-647P1 via SOS1 was measured using an HTRF® compatible reader (PHERAstar, BMG Labtech, Germany) with Ex / Em=(337 / 665; 337 / 620), and the compound concentration with no SOS1 reaction or the highest concentration was used as a blank. IC50 was measured using a sigmoidal dose response curve (variation slope, var i ab 1 e slope).

[0218] The inhibitory effect of the interaction between KRAS(G12C) protein and S0S2 protein was measured using the same method as above, except that KRAS(G12C) was used instead of KRAS(G12C, G12D, G12V) protein and S0S2 protein was used instead of S0S1 protein.

[0219] 【Table 11

[0220] Target inhibition (IC50, nM) KRAS(G12C)- KRAS(G12D)- KRAS(G12V)- KRAS(G12C)- S0S1 S0S1 S0S2 compounds

[0221] 24.48 16.39 33.48 >10 , 000 A

[0222]

[0223] As shown in Table 1 above, it was confirmed that compound A exhibits inhibitory activity against the interaction between KRAS (G12C, G12D, G12V) proteins and S0S1 proteins, and that it does not inhibit the interaction between KRAS (CG12C) and S0S2 (IC50 >10,000 nM). Through the above results, it was confirmed that compound A of the present invention is a selective inhibitor of S0S1 and exhibits pan-KRAS inhibitory activity.

[0224] <Experimental Example 2> Evaluation of 3D Spheroid Assay

[0225] The cell growth inhibitory efficacy of the S0S1 inhibitory compound according to the present invention (Example compound; Compound A) on various KRAS mutant cancer cell lines was evaluated.

[0226] Three KRAS mutant cancer cell lines (MIA PaCa-2, Calu-1, AsPC-1) were seeded at a density of 1,000 cells / well in 96-well ultra-low adsorption culture plates. The cells were cultured under conditions of 37 °C and 5% CO2 for 3 to 4 days, during which the solution was repeatedly replaced with one containing the compound of the example and fresh medium. Culture was then continued until day 7 to form 3D cell spheroids. Subsequently, 50 M Cell Titer-Gio 3D reagent was added, and cell lysis was performed for 5 minutes. After stabilization for 25 minutes, luminescence was measured, and the IC50 was determined using GraphPad Prism 7 software.

[0227] As shown in Figure 1, it was confirmed that compound A inhibits cell growth in a concentration-dependent manner in various KRAS variant cancer (G12C, G12D) cell lines.

[0228] <Experimental Example 3> Ex vivo PK evaluation

[0229] To evaluate the PK profile of the S0S1 inhibitor compound according to the present invention (Example compound; Compound A), an ex vivo PK measurement experiment was performed by administering Compound A of the present invention as monotherapy to a KRAS(G12C) mutation pancreatic cancer MIA PaCa-2 xenograft model. Specifically, 50 mg / kg of Compound A was orally administered once daily, five times a week for 24 days to a KRAS(G12C) mutation pancreatic cancer MIA PaCa-2 xenograft model. The experiment was conducted with 31 mice, and on day 24, three mice were selected at each time point from among those whose tumors were close to the average of each group to evaluate their blood PK profiles.

[0230] To evaluate the PK profile in blood over time, collected blood was separated into plasma, and the plasma and tumor tissue were stored in an ultra-low temperature freezer at -70°C until analysis. To analyze the concentration of compound A in mouse plasma and tumor tissue, the samples were pretreated by the protein precipitation method, and then compound A was quantitatively analyzed using LC-MS / MS.

[0231] When compound A was orally administered to a KRASCG12C mutation pancreatic cancer MIA PaCa-2 xenograft mouse model, the drug was detected in the plasma for 8 hours and in the tumor for 24 hours, showing a drug distribution nearly twice as high in the tumor compared to the plasma (Fig. 2). Through the above results, it was confirmed that when the compound of the embodiment of the present invention is orally administered, it exhibits excellent drug distribution in both the plasma and the tumor, and in particular, shows a high drug concentration in the tumor for a long time compared to the plasma.

[0232] <Experimental Example 4> Evaluation of Cell Growth Inhibitory Ability of Combination Therapy with KRAS(G12C) Inhibitor and S0S1 Inhibitor on KRAS(G12C) Mutant Cell Lines

[0233] To evaluate the cell growth inhibitory efficacy of the combination therapy of a KRAS(G12C) inhibitor and an S0S1 inhibitor on KRAS(G12C) mutant cell lines, a cell viability assay was performed.

[0234] The FDA-approved KRAS(G12C) inhibitors used were 'Adagrasib' and 'Sotorasib'. 'Adagrasib' (Cas No. 2326521-71-3) was purchased from IKSChem for research purposes, and 'Sotorasib' (Cas No. 2296729-00-3) was purchased from Habotech for research purposes.

[0235] Three KRASCG12C) mutant cancer human cell lines (MIA PaCa-2, pancreatic cancer; NCI-H1373, non-small cell lung cancer; SW837, colorectal cancer) were cultured in media suitable for their respective growth conditions containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS). After culturing for 24 hours in 96-cell culture plates with an optimal cell count per cell in each medium, the cells were cultured for 3 days at 37 °C and 5% CO2 conditions in cell culture media containing concentration-dependent combinations of the KRASCG12C) inhibitor and compound A of the present invention or dimethyl sulfoxide (DMSO) vehicle (Table 2).

[0236] [Table 2]

[0237] Cell count in culture medium of KRAS(G12C) mutant cancer cell line

[0238] MIA PaCa-2 DMEM 3 X 10 3 cel ls / wel l NCI-H1373 RPMI-1640 3 3 cel ls / wel l SW837 RPMI-1640 2.5 4 cel ls / wel l

[0239]

[0240] To confirm cell viability, a solution of cell counting assay kit-8 (CCK-8, Dojindo) was added to each of the cultured cells and reacted for 4 hours at 37 °C under 5% CO2 conditions, after which the absorbance was measured at the 450 band. The cell growth inhibitory efficacy of combination therapy according to concentration combinations of a KRASCG12C inhibitor and Compound A of the present invention was evaluated using the HSA synergy score, which was calculated using the SynergyFinder program. The synergistic effect on cell growth inhibition was evaluated based on the following criteria:

[0241] • HSA Synergy Score > 10: Increase (synergist ic)

[0242] • -10 < HSA Synergy Score < 10: Additive

[0243] • HSA Synergy Score < -10: Antagonistic

[0244] The results of evaluating the cell growth inhibitory efficacy of combination therapy of a KRASCG12C inhibitor and compound A of the present invention for each cell line using the HSA synergy score are shown in Figures 3 to 5.

[0245] As a result of co-administration of compound A with the KRAS G12C inhibitors Adagrasib or Sotorasib in MIA PaCa-2 cell lines (pancreatic cancer), the portion with an HSA synergy score greater than 10 was confirmed for each combination with Adagrasib or Sotorasib, confirming that there is a synergistic effect on inhibiting cancer cell growth when compound A is co-administered with a KRAS G12C inhibitor (Fig. 3).

[0246] As a result of co-administration of compound A with the KRAS G12C inhibitors Adagrasib or Sotorasib in the NCI-H1373 cell line (non-small cell lung cancer), the portion with an HSA synergy score greater than 10 was confirmed for each combination with Adagrasib or Sotorasib, confirming that there is a synergistic effect on inhibiting cancer cell growth when compound A is co-administered with a KRAS G12C inhibitor (Fig. 4).

[0247] As a result of co-administration of Compound A with the KRAS G12C inhibitors Adagrasib or Sotorasib in SW837 cell lines (colorectal cancer), the HSA synergy score exceeding 10 was confirmed for each co-administration with Adagrasib or Sotorasib, confirming that there is a synergistic effect on inhibiting cancer cell growth when Compound A is co-administered with a KRAS G12C inhibitor (Fig. 5). Therefore, the co-administration of Compound A and the KRAS G12C inhibitor of the present invention exhibits a high synergistic effect on inhibiting the growth of KRAS(G12C) mutant cancer cells. Accordingly, it can be effectively used as a cancer treatment.

[0248] <Experimental Example 5> Evaluation of Cell Growth Inhibitory Ability of Combination Therapy of KRAS(G12D) Inhibitor and S0S1 Inhibitor on KRAS(G12D) Mutant Cell Lines

[0249] To evaluate the cell growth inhibitory efficacy of the combination therapy of a KRAS(G12D) inhibitor and an S0S1 inhibitor on KRAS(G12D) mutant cell lines, a cell viability assay was performed.

[0250] The KRAS(G12D) inhibitor 'MRTX1133' (Cas No. 2621928-55-8) was purchased from Angene for research purposes and used.

[0251] Human cell lines of KRASCG12D) mutant cancer (AsPC-1, pancreatic cancer) were cultured in media suitable for growth conditions containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS). After culturing the optimal number of cells per cell in each medium in 96-cell culture plates for 24 hours, the cells were cultured for 3 days at 37 °C and 5% CO2 conditions in cell culture media containing concentration-dependent combinations of the KRASCG12D) inhibitor and compound A of the present invention or dimethyl sulfoxide (DMSO) vehicle (Table 3).

[0252] [Table 3]

[0253] Cell count in culture medium of KRAS(G12D) mutant cancer cell line

[0254]

[0255] AsPC-1 RPMI-1640 5 X 10 3 cel ls / wel l

[0256]

[0257] To confirm cell viability, the cell counting assay kit-8 (CCK-8, Do j indo) solution was added to the above cells and incubated for 4 hours under 37 °C and 5% CO2 conditions, after which absorbance was measured at 450 counts. The cell growth inhibitory efficacy of the combination therapy of each compound was evaluated using the HSA synergy score, which was calculated using the SynergyFinder program. The synergistic effect on cell growth inhibition was evaluated based on the following criteria:

[0258] • HSA Synergy Score > 10: Increase (synergist ic)

[0259] - -10 < HSA Synergy Score < 10: Additive

[0260] • HSA Synergy Score < -10: Antagonistic

[0261] As shown in Figure 6, when compound A and the KRAS G12D inhibitor MRTX1133 were administered in combination to AsPC-1 cell lines (pancreatic cancer), a region with an HSA synergy score greater than 10 was identified, confirming that there is a synergistic effect on inhibiting cancer cell growth when compound A and the KRAS G12D inhibitor are administered in combination.

[0262] Therefore, the combined administration of compound A of the present invention and a KRAS G12D inhibitor exhibits a high synergistic effect in inhibiting the growth of KRASCG12D-mutated cancer cells. Accordingly, it can be effectively used as a cancer treatment agent.

[0263] <Experimental Example 6> Evaluation of tumor growth inhibition ability of KRAS(G12C) inhibitor and S0S1 inhibitor monotherapy and combination therapy in a KRAS(G12C) mutation xenograft model. The tumor growth inhibition efficacy was confirmed in a KRASCG12C) mutation xenograft model using monotherapy of the S0S1 inhibitor compound (Compound A) according to the present invention and combination therapy with a KRASCG12C) inhibitor.

[0264] The FDA-approved KRAS(G12C) inhibitors 'Adagrasib' and 'Sotorasib' were used, and a xenograft model study was conducted to evaluate the tumor growth inhibitory efficacy.

[0265] (1) KRAS(G12C) mutation pancreatic cancer (MIA PaCa-2) xenograft model study

[0266] 5–6 week old female athymic nude mice were used as experimental animals. Human pancreatic cancer cells, MIA PaCa-2, were mixed with PBS and Matri Gel in a 1:1 ratio and injected subcutaneously into the right lateral side of the mice using 1x1O. 7 Cells / 200 mcg / mice were transplanted. When the tumor size reached approximately 100–200 mnF, mice were grouped according to the average tumor size, with 8–9 mice used per group. All test substances (KRAS(G12C) inhibitor monotherapy group; Compound A monotherapy group; KRAS(G12C) inhibitor + Compound A combination therapy group; Vehicle therapy group) were administered orally once daily, five times a week. In the combination therapy groups, the KRAS(G12C) inhibitor was administered first, followed by sequential administration of Compound A. Tumor size was measured twice a week using a caliper to determine the length and width of the tumor, and the tumor volume (mm²) 3) is 0.5 x (major axis) x (minor axis) 2 It was calculated as follows.

[0267] The study of the KRAS(G12C) mutant pancreatic cancer (MIA PaCa-2) xenograft model was evaluated by measuring the tumor growth inhibition rate, changes in tumor weight, and changes in mouse body weight in the xenograft model. 1) The results of the evaluation of the tumor growth inhibition rate in the monotherapy and combination therapy xenograft models of adagrasib and Compound A are shown in Figure 7. 24 days after administration of the test substance, the adagrasib (KRAS(G12C) inhibitor) monotherapy group and the adagrasib + Compound A combination therapy group showed significant tumor growth inhibition compared to the vehicle administration group. The combination therapy group of Adagrasib + Compound A showed significant tumor growth inhibition when compared to the Adagrasib monotherapy group and the Compound A monotherapy group, respectively, and a synergistic effect between the KRASCG12C inhibitor and Compound A was confirmed.

[0268] The results of measuring changes in tumor weight in a xenograft model are shown in Figure 8. After 24 days of administering the test substance, the group administered adagrasib alone and the group administered adagrasib + compound A showed significant inhibition of tumor weight compared to the vehicle administration group. The group administered adagrasib + compound A also showed significant inhibition of weight growth when compared to the group administered adagrasib alone and the group administered compound A alone, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and compound A.

[0269] As shown in Figure 9, all test substance administration groups showed body weight changes similar to the vehicle administration groups without body weight loss caused by the test substance. Accordingly, it was confirmed that all test substances were non-toxic to mice.

[0270] 2) The results of evaluating the tumor growth inhibition rates in xenograft models of sotorasib and compound A monotherapy and combination therapy are shown in Fig. 10. After 24 days of administering the test substance, the sotorasib + compound A combination therapy group showed significant tumor growth inhibition compared to the vehicle therapy group. The sotorasib + compound A combination therapy group showed significant tumor growth inhibition when compared to the sotorasib monotherapy group and the compound A monotherapy group, respectively, and a synergistic effect between the KRASCG12C inhibitor and compound A was confirmed.

[0271] The results of measuring changes in tumor weight in a xenograft model are shown in Fig. 11. After 24 days of administering the test substance, the combination therapy group of Sotorasib + Compound A showed significant inhibition of tumor weight compared to the vehicle therapy group. The combination therapy group of Sotorasib + Compound A also showed significant inhibition of weight growth when compared to the Sotorasib monotherapy group and the Compound A monotherapy group, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and Compound A.

[0272] As shown in Fig. 12, all test substance administration groups showed a change in body weight similar to the vehicle administration group without drug-induced weight loss. Accordingly, it was confirmed that all test substances were non-toxic to mice.

[0273] Based on the above results, it was confirmed that the combined administration of a KRAS(G12C) inhibitor and Compound A exhibited superior synergistic effects in inhibiting tumor growth and reducing tumor weight in a KRAS(G12C) mutant pancreatic cancer (MIA PaCa-2) xenograft model compared to the monotherapy of the KRAS(G12C) inhibitor and Compound A, respectively. Additionally, no significant decrease in mouse body weight was observed, confirming that there was no toxicity. (2) Study of a KRAS(G12C) mutant lung cancer (NCI-H1373) xenograft model

[0274] 5–6 week old female athymic nude mice were used as experimental animals. 5 x 10⁶ human non-small cell lung cancer cells (NCI-H1373) were mixed with PBS and Matrigel in a 1:1 ratio and injected subcutaneously into the right lateral side of the mice. 6 Cells were transplanted at a rate of 100 m / mouse. When the tumor size reached approximately 100–200 mnf, mice were grouped according to the average tumor size, with 9–10 mice used per group. All test substances were administered orally once daily, five times a week. In the combination therapy group, the KRASCG12C inhibitor was administered first, followed by the sequential administration of Compound A. Tumor size was measured twice a week using a caliper to determine the length and width, and the tumor volume (mm²) 3 ) is 0.5 x (major axis) x (minor axis) 2 It was calculated as follows.

[0275] The study of the KRASCG12C) mutation lung cancer (NCI-H1373) xenograft model was evaluated by measuring the tumor growth inhibition rate, changes in tumor weight, and changes in mouse body weight in the xenograft model.

[0276] 1) Monotherapy and combination therapy of Adagrasib and Compound A

[0277] The results of the evaluation of tumor growth inhibition rates in a xenograft model are shown in Fig. 13. After 22 days of administration of the test substance, the group administered compound A alone, the group administered adagrasib alone, and the group administered adagrasib + compound A showed significant tumor growth inhibition compared to the vehicle administration group. The group administered adagrasib + compound A also showed significant tumor growth inhibition when compared to the group administered adagrasib alone and the group administered compound A alone, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and compound A.

[0278] The results of measuring changes in tumor weight in a xenograft model are shown in Fig. 14. After 22 days of administering the test substance, the group administered compound A alone, the group administered adagrasib alone, and the group administered adagrasib + compound A showed significant inhibition of tumor weight compared to the vehicle administration group. The group administered adagrasib + compound A also showed significant inhibition of weight growth when compared to the group administered adagrasib alone and the group administered compound A alone, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and compound A.

[0279] As shown in Fig. 15, all test substance administration groups showed body weight changes similar to the vehicle administration groups without body weight loss caused by the test substance. Accordingly, it was confirmed that all test substances were not toxic to mice.

[0280] 2) Sotorasib and Compound A alone and in combination

[0281] The results of the evaluation of tumor growth inhibition rates in the xenograft model are shown in Fig. 16. Twenty-two days after administration of the test substance, the groups administered Compound A alone, Sotorasib alone, and the combination of Sotorasib and Compound A showed significant tumor growth inhibition compared to the vehicle administration group. The combination of Sotorasib and Compound A also showed significant tumor growth inhibition when compared to the Sotorasib alone group and the Compound A alone group, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and Compound A. The results of measuring changes in tumor weight in the xenograft model are shown in Fig. 17. Twenty-two days after administration of the test substance, the groups administered Compound A alone, Sotorasib alone, and the combination of Sotorasib and Compound A showed significant tumor weight inhibition compared to the vehicle administration group. The combination therapy group of Sotorasib + Compound A showed significant weight growth inhibition when compared to the Sotorasib monotherapy group and the Compound A monotherapy group, respectively, and the synergistic effect of the KRASCG12C inhibitor and Compound A was confirmed.

[0282] As shown in Fig. 18, all test substance administration groups showed body weight changes similar to the vehicle administration groups without body weight loss caused by the test substance. Accordingly, it was confirmed that all test substances were not toxic to mice.

[0283] Through the above results, it was confirmed that the combined administration of a KRASCG12C inhibitor and compound A showed excellent synergistic effects in inhibiting tumor growth and reducing tumor weight in a KRAS(G12C) mutation lung cancer (NCI-H1373) xenograft model compared to the single administration of the KRASCG12C inhibitor and compound A, respectively, and it was confirmed that there was no toxicity as no significant decrease in mouse body weight was observed.

[0284] (3) KRAS(G12C) mutation colorectal cancer (SW837) xenograft model study

[0285] 5–6 week old female athymic nude mice were used as experimental animals. Human colorectal cancer cells, SW837, were mixed with PBS and Matri Gel in a 1:1 ratio and injected 5 x 10⁻¹⁰ into the subcutaneous tissue of the right lateral side of the mice. 6 Cells were transplanted at a rate of 200 m / mouse. When the tumor size reached approximately 100–200 mF, mice were grouped according to the average tumor size, with 10 mice per group. All test substances were administered orally once daily, five times a week. In the combination therapy group, the KRASCG12C inhibitor was administered first, followed by the sequential administration of Compound A. Tumor size was measured twice a week using a caliper to determine the length and width, and the tumor volume (mm²) 3 ) is 0.5 x (major axis) x (minor axis) 2 It was calculated as follows.

[0286] The study of the KRASCG12C) variant colorectal cancer (SW837) xenograft model was evaluated by measuring the tumor growth inhibition rate, changes in tumor weight, and changes in mouse body weight in the xenograft model.

[0287] 1) Monotherapy and combination therapy of Adagrasib and Compound A

[0288] The results of the evaluation of tumor growth inhibition rates in a xenograft model are shown in Fig. 19. 18 days after administration of the test substance, the combination therapy group of Adagrasib + Compound A showed significant tumor growth inhibition compared to the vehicle therapy group. The combination therapy group of Adagrasib + Compound A also showed significant tumor growth inhibition when compared to the Adagrasib monotherapy group and the Compound A monotherapy group, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and Compound A.

[0289] The results of measuring changes in tumor weight in a xenograft model are shown in Fig. 20. Eighteen days after administration of the test substance, the group administered with the combination of Adagrasib and Compound A showed significant inhibition of tumor weight compared to the vehicle administration group. The group administered with the combination of Adagrasib and Compound A also showed significant inhibition of weight growth when compared to the Adagrasib monotherapy group and the Compound A monotherapy group, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and Compound A. As shown in Fig. 21, all test substance administration groups showed weight changes similar to the vehicle administration group without weight loss caused by the test substance. Accordingly, it was confirmed that all test substances were non-toxic to mice.

[0290] 2) Monotherapy and combination therapy of Sotorasib and Compound A

[0291] The results of the evaluation of tumor growth inhibition rates in a xenograft model are shown in Fig. 22. After 22 days of administering the test substance, the group administered compound A alone, the group administered sotorasib alone, and the group administered sotorasib + compound A showed significant tumor growth inhibition compared to the vehicle administration group. The group administered sotorasib + compound A also showed significant tumor growth inhibition when compared to the sotorasib alone group and the group administered compound A alone, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and compound A.

[0292] The results of measuring changes in tumor weight in a xenograft model are shown in Fig. 23. 22 days after administration of the test substance, the Sotorasib monotherapy group and the Sotorasib + Compound A combination therapy group showed significant inhibition of tumor weight compared to the vehicle therapy group. The Sotorasib + Compound A combination therapy group also showed significant inhibition of weight growth when compared to the Sotorasib monotherapy group and the Compound A monotherapy group, respectively, confirming the synergistic effect of the KRASCG12C inhibitor and Compound A.

[0293] As shown in Fig. 24, all test substance administration groups showed a change in body weight similar to the vehicle administration group without any weight loss caused by the test substance. Accordingly, it was confirmed that all test substances were non-toxic to mice. Through the above results, it was confirmed that the combined administration of a KRAS(G12C) inhibitor and Compound A exhibited superior synergistic effects in inhibiting tumor growth and reducing tumor weight in a KRAS(G12C) mutation colorectal cancer (SW837) xenograft model compared to the single administration of the KRAS(G12C) inhibitor and Compound A, respectively, and it was confirmed that there was no toxicity as no significant decrease in mouse body weight was observed.

[0294] <Experimental Example 7> Evaluation of Tumor Growth Inhibitory Ability of KRAS(G12D) Inhibitors and S0S1 Inhibitors in Monotherapy and Combination Therapy in a KRAS(G12D) Mutation Xenograft Model

[0295] The tumor growth inhibitory efficacy of the S0S1 inhibitory compound (Compound A) according to the present invention was confirmed in a KRASCG12D) mutation xenograft model in combination therapy with a KRASCG12D) inhibitor.

[0296] The KRASCG12D inhibitor 'MRTX1133' was used, and a xenograft model study was performed to evaluate the tumor growth inhibitory efficacy.

[0297] 5–6 week old female athymic nude mice were used as experimental animals. Human pancreatic cancer cells, AsPC-1, were mixed with PBS and Matri Gel in a 1:1 ratio and injected subcutaneously into the right lateral side of the mice using 1x1O. 7Cells were transplanted into 200 mice. When the tumor size reached approximately 100–200 mF, mice were grouped according to the average tumor size, with 10 mice used per group. MRTX1133 was administered intraperitoneally twice daily, five times a week, while all other test substances were administered orally once daily, five times a week. In the combination therapy group, the KRASCG12D inhibitor was administered first, followed by sequential administration of Compound A. Tumor size was measured twice a week using a caliper to determine the length and width of the tumor, and the tumor volume (mm²) 3 ) is 0.5 x (major axis) x (minor axis) 2 It was calculated as follows.

[0298] Studies on the KRASCG12D) variant pancreatic cancer (AsPC-1) xenograft model were evaluated by measuring the tumor growth inhibition rate, changes in tumor weight, and changes in mouse body weight in the xenograft model.

[0299] The results of the evaluation of tumor growth inhibition rates in a xenograft model are shown in Fig. 25. After 24 days of drug administration, the MRTX1133 monotherapy group and the MRTX1133 + compound A combination therapy group showed significant tumor growth inhibition compared to the vehicle therapy group. The MRTX1133 + compound A combination therapy group also showed significant tumor growth inhibition when compared to the MRTX1133 monotherapy group and the compound A monotherapy group, respectively, confirming the synergistic effect of the KRASCG12D inhibitor and compound A.

[0300] The results of measuring changes in tumor weight in a xenograft model are shown in Fig. 26. After 24 days of administering the test substance, the MRTX1133 monotherapy group and the MRTX1133 + compound A combination therapy group showed significant inhibition of tumor weight compared to the vehicle therapy group. The MRTX1133 + compound A combination therapy group also showed significant inhibition of weight growth when compared to the MRTX1133 monotherapy group and the compound A monotherapy group, respectively, confirming the synergistic effect of the KRASCG12D inhibitor and compound A.

[0301] As shown in Fig. 27, all test substance administration groups showed a change in body weight similar to the vehicle administration group without any weight loss caused by the test substance. Accordingly, it was confirmed that all test substances were non-toxic to mice. Through these results, it was confirmed that the combined administration of a KRAS(G12D) inhibitor and Compound A exhibited superior synergistic effects in inhibiting tumor growth and reducing tumor weight in a KRAS(G12D) mutation pancreatic cancer (AsPC-1) xenograft model compared to the single administration of the KRAS(G12D) inhibitor and Compound A, respectively, and it was confirmed that there was no toxicity as no significant decrease in mouse body weight was observed.

Claims

【Scope of Claim】 【Claim 11 A pharmaceutical composition for the prevention or treatment of cancer comprising a compound represented by the following chemical formula I or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered in combination with a KRAS inhibitor: [Chemical Formula I]

2. In claim 1, the pharmaceutically acceptable salt of the compound represented by chemical formula I is a hydrochloric acid salt, a pharmaceutical composition.

3. In paragraph 2, the above-mentioned hydrochloric acid salt is a monohydrochloric acid salt, 1.5-hydrochloric acid salt, or dihydrochloric acid salt, a pharmaceutical composition. 【claim Paragraph 4] In paragraph 1, the KRAS inhibitor is a KRAS G12C inhibitor, a KRAS G12D inhibitor, or is a pharmaceutical composition that is a KRAS G12V inhibitor.

5. In claim 4, the KRAS G12C inhibitor is one or more selected from the group consisting of Sotoras ib, Adagras ib, Olomoras ib, Divaras ib, Gleciras ib, Calderas ib, JNJ-74699157, ZG-19018, YL-15293, ICP-915, BI-1823911, BEBT-607, ERAS-3490, and BPI-421286, a pharmaceutical composition.

6. In claim 4, the pharmaceutical composition wherein the KRAS G12D inhibitor is one or more selected from the group consisting of MRTX1133, Zoldonrasib (Zo ldonras ib), HRS-4642, INCB161734, QTX3034, AZD0022, and BI-KRASG12D3. 【claim Paragraph 7] A pharmaceutical composition according to claim 1 comprising a compound represented by chemical formula I or a pharmaceutically acceptable salt thereof; and a KRAS inhibitor each formulated and administered simultaneously or sequentially. 【claim Paragraph 8] A pharmaceutical composition according to claim 1, comprising a compound represented by chemical formula I or a pharmaceutically acceptable salt thereof; and a pharmaceutical composition wherein the KRAS inhibitor is each included in the form of a separate formulation or a mixed form. 【Claim this In claim 1, the cancer is a pharmaceutical composition comprising a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation.

10. A pharmaceutical composition according to claim 1, wherein the cancer is a cancer containing a KRAS G12C mutation, and the KRAS inhibitor is a KRAS G12C inhibitor.

11. A pharmaceutical composition according to claim 1, wherein the cancer is a cancer containing a KRAS G12D mutation, and the KRAS inhibitor is a KRAS G12D inhibitor.

12. A pharmaceutical composition according to claim 1, wherein the cancer is one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma, and solid tumors.

13. A pharmaceutical composition according to claim 1, wherein the cancer is one or more selected from the group consisting of pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, colorectal cancer containing a KRAS G12C mutation, pancreatic cancer containing a KRAS G12D mutation, non-small cell lung cancer containing a KRAS G12D mutation, colorectal cancer containing a KRAS G12D mutation, pancreatic cancer containing a KRAS G12V mutation, non-small cell lung cancer containing a KRAS G12V mutation, and colorectal cancer containing a KRAS G12V mutation.

14. (i) A first pharmaceutical composition comprising a compound represented by the following chemical formula I or a pharmaceutically acceptable salt thereof: [Chemical Formula I] % ( ii ) A second pharmaceutical composition comprising a KRAS inhibitor A combination of pharmaceutical compositions for concomitant administration for the prevention or treatment of cancer, including

15. In paragraph 14, the pharmaceutically acceptable salt of the compound represented by chemical formula I is a combination of hydrochloric acid salts.

16. In paragraph 15, the above hydrochloric acid salt is a combination of monohydrochloric acid, 1.5-hydrochloric acid, or dihydrochloric acid salt.

17. In paragraph 14, the KRAS inhibitor is a combination of a KRAS G12C inhibitor, a KRAS G12D inhibitor, or a KRAS G12V inhibitor.

18. In paragraph 17, the KRAS G12C inhibitor is a combination of one or more selected from the group consisting of Sotoras ib, Adagras ib, Olomoras ib, Divaras ib, Gleciras ib, Calderas ib, JNJ-74699157, ZG-19018, YL-15293, ICP-915, BI-1823911, BEBT-607, ERAS-3490, and BPI-421286. 【Claim 1 is In paragraph 17, the KRAS G12D inhibitor is a combination of one or more selected from the group consisting of MRTX1133, Zoldonrasib, HRS-4642, INCB161734, QTX3034, AZD0022, and BI-KRASG12D3.

20. In paragraph 14, the cancer is a combination comprising a KRAS G12C mutation, a KRAS G12D mutation, or a KRAS G12V mutation.

21. In paragraph 14, cancer is a combination of one or more selected from the group consisting of lung cancer, lung adenocarcinoma, non-small cell lung cancer, squamous cell lung cancer, pancreatic cancer, pancreatic adenocarcinoma, colorectal cancer, colorectal cancer, colorectal adenocarcinoma, rectal cancer, rectal cancer adenocarcinoma, appendiceal cancer, small intestine cancer, esophageal cancer, bladder cancer, ovarian cancer, uterine cancer, endometrial cancer, liver cancer, hepatobiliary cancer, cholangiocarcinoma, and solid tumors.

22. In claim 14, the cancer is a combination of one or more selected from the group consisting of pancreatic cancer containing a KRAS G12C mutation, non-small cell lung cancer containing a KRAS G12C mutation, colorectal cancer containing a KRAS G12C mutation, pancreatic cancer containing a KRAS G12D mutation, non-small cell lung cancer containing a KRAS G12D mutation, colorectal cancer containing a KRAS G12D mutation, pancreatic cancer containing a KRAS G12V mutation, non-small cell lung cancer containing a KRAS G12V mutation, and colorectal cancer containing a KRAS G12V mutation.

23. (i) A first pharmaceutical composition comprising a compound represented by the following chemical formula I or a pharmaceutically acceptable salt thereof: [Chemical Formula I] (ii) A second pharmaceutical composition comprising a KRAS inhibitor A kit for combination administration for the prevention or treatment of cancer containing

24. Use of a pharmaceutical composition according to Article 1 for the prevention or treatment of cancer.

25. Use of the combination of substances pursuant to Paragraph 14 for the prevention or treatment of cancer.

26. Use of the kit for the prevention or treatment of cancer pursuant to Paragraph 23.

27. Use of a pharmaceutical composition according to claim 1 for the manufacture of a drug for the prevention or treatment of cancer.

28. Use of the combination pursuant to Paragraph 14 for the manufacture of a drug for the prevention or treatment of cancer 【Claim 2 Use of the kit according to paragraph 23 for the manufacture of a drug for the prevention or treatment of cancer.

30. A method for the prevention or treatment of cancer comprising the step of administering a pharmaceutical composition according to claim 1 to an individual in need thereof.

31. A method for the prevention or treatment of cancer comprising the step of administering the combination according to Paragraph 14 to an individual in need thereof.

32. A method for the prevention or treatment of cancer comprising the step of administering a kit according to paragraph 23 to an individual in need thereof.

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