Combination Therapies for the Treatment of Cancer

Abstract

The present disclosure provides methods of treating cancer using combination therapy of an SHP2 inhibitor and a KRAS G12C inhibitor.

Description

[Technical Field]

[0001] cross reference This application claims the advantages of U.S. Provisional Patent Application No. 63 / 124,669 filed on 11 December 2020, U.S. Provisional Patent Application No. 63 / 214,736 filed on 24 June 2021, U.S. Provisional Patent Application No. 63 / 277,555 filed on 9 November 2021, and U.S. Provisional Patent Application No. 63 / 283,035 filed on 24 November 2021, which are each incorporated herein by reference in their entirety. [Background technology]

[0002] Src homology-2 phosphatase (SHP2) is a non-receptor protein phosphatase that is ubiquitously expressed in various tissues and cell types (see the overview below, Tajan M etal., Eur J Med Genet 2016 58(10):509-25, Grossmann KS etal., Adv Cancer Res 2010 106:53-89). SHP2 consists of two Src homology 2 (N-SH2 and C-SH2) domains at its NH2 terminus, a catalytic PTP (protein tyrosine phosphatase) domain, and a regulatory C-terminal tail. In the ground state, intermolecular interactions between the SH2 domain and the PTP domain prevent substrate access to the catalytic pocket, maintaining SHP2 in a closed, self-inhibitory conformation. In response to stimulation, the SHP2-activating protein, which contains a phosphorylated tyrosine motif, binds to the SH2 domain, leading to exposure of the active site and enzymatic activation of SHP2. [Overview of the project]

[0003] The embodiments disclosed herein generally relate to compositions and methods relating to combination therapies for treating cancer that utilize SHP2 inhibitors in combination with KRAS G12C inhibitors, while providing an unexpected degree of synergy.

[0004] SHP2 plays a crucial role in fundamental cellular functions, including proliferation, differentiation, cell cycle maintenance, and motility. By dephosphorylating its associated signaling molecules, SHP2 modulates multiple intracellular signaling pathways in response to a wide range of growth factors, cytokines, and hormones. Cellular signaling processes involving SHP2 include the RAS-MAPK (mitogen-activated protein kinase), PI3K (phosphoinositol 3-kinase)-AKT, and JAK-STAT pathways.

[0005] SHP2 also plays a signal-enhancing role in this pathway, acting downstream of RTKs and upstream of RAS. One common mechanism of resistance to pharmacological inhibition of MAPK signaling involves RTK activation, which promotes the reactivation of MAPK signaling. RTK activation recruits SHP2 via direct binding and via adapter proteins. These interactions result in the conversion of SHP2 from a closed (inactive) conformation to an open (active) conformation. SHP2 is a key promoter of RAS signaling reactivation, evading pharmacological inhibition in both primary and secondary resistance. Inhibition of SHP2 often achieves an overall attenuation effect of stream RTK signaling that drives oncogenic signaling and adaptive tumor escape (see Prahallad, A. et al., Cell Reports 12, 1978-1985 (2015), Chen YN, Nature 535, 148-152 (2016)), and all of its teachings, including but not limited to all methods, compounds, compositions, data, etc., for use in any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety.

[0006] In addition to SHP2, the RAS-MAPK signaling pathway includes the RAS family of proteins. This family includes three related GTPases (K-, N-, and HRAS) that play a role in the signaling pathway. In particular, KRAS is known to have numerous mutations that indicate oncogenic status. KRAS variants, such as mutations occurring at amino acid residue 12 (i.e., G12X), are generally known to cause cancer. For example, the G12C mutation occurs in about 13% of NSCLC patients, as well as in 1%–3% of colorectal cancers and solid tumors.

[0007] In a first aspect, the Disclosure relates to a method for treating a subject having cancer, wherein the subject is given a therapeutically effective amount of Formula I:

[0008] [ka] The present invention provides a method comprising the step of administering a compound or a pharmaceutically acceptable salt thereof in combination with a KRAS inhibitor.

[0009] In some embodiments, the cancer contains the KRAS G12C mutation.

[0010] In some embodiments, the cancer is lung cancer.

[0011] In some embodiments, the cancer is non-small cell lung cancer.

[0012] In some embodiments, the cancer is esophageal cancer.

[0013] In some embodiments, the cancer is pancreatic ductal adenocarcinoma (PDAC).

[0014] In some embodiments, inhibitors of mutant KRAS are selected from the group consisting of AMG510 (sotorasib, LUMAKRAS®), MRTX849 (adagrasib), ARS-3248, GDC-6036, BI 1701963, tipifarnib, and BBP-454.

[0015] In some embodiments, the KRAS inhibitor is AMG510.

[0016] In some embodiments, the KRAS inhibitor is MRTX849.

[0017] In some embodiments, the KRAS inhibitor is ARS-3248.

[0018] In some embodiments, the KRAS inhibitor is BI 1701963.

[0019] In some embodiments, the method includes administering a third MAPK pathway inhibitor.

[0020] In some embodiments, the administration is oral.

[0021] In some embodiments, the dosage of the compound of formula I is in the range of 20 mg to 400 mg per day.

[0022] In some embodiments, the dosage of the KRAS inhibitor is in the range of 1 mg to 1,000 mg per day.

[0023] In a second aspect, the present disclosure provides a method for treating lung cancer or esophageal cancer in a subject, the method comprising orally administering to the subject a therapeutically effective amount of a compound of formula I:

[0024]

Chemical formula

[0025] In some embodiments, the compound of formula I is administered once or twice a day.

[0026] In some embodiments, AMG510 is administered once or twice a day.

[0027] In some embodiments, the subject is human.

[0028] In a third aspect, the disclosure relates to a method for treating a target cancer, wherein the target is given a therapeutically effective amount of formula I:

[0029] [ka] The present invention provides a method comprising the step of orally administering a compound or a pharmaceutically acceptable salt thereof in combination with MRTX849.

[0030] In some embodiments, the cancer is lung cancer, colorectal cancer, esophageal cancer, or breast cancer.

[0031] In some embodiments, the cancer is pancreatic ductal adenocarcinoma (PDAC).

[0032] In some embodiments, the compound of formula I is administered once or twice a day.

[0033] In some embodiments, MRTX849 is administered once or twice daily.

[0034] In some embodiments, the subject is human.

[0035] In some embodiments, a kit is provided comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a KRAS inhibitor.

[0036] In some embodiments, the compound of formula I and the KRAS inhibitor are located in separate packages.

[0037] In some embodiments, the kit further includes instructions for administering the contents of the kit to a subject for the treatment of cancer.

[0038] In some embodiments, the KRAS inhibitor is one or more of the following: AMG510, MRTX849, ARS-3248, GDC-6036, BI 1701963, tipifarnib, and BBP-454.

[0039] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as an oral composition.

[0040] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once or twice daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered twice daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a continuous 28-day cycle.

[0041] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in an amount of about 10 mg to about 140 mg.

[0042] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a three-week cycle comprising administering the compound for two weeks, followed by a one-week period without administration of the compound.

[0043] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a four-week cycle comprising administering the compound for three weeks, followed by a one-week period without administration of the compound.

[0044] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of 6 weeks. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of 8 weeks.

[0045] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered three times a week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first, third, and fifth days of the week.

[0046] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered four times a week.

[0047] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a three-week cycle comprising administering the compound for two weeks, followed by a one-week period without administration of the compound.

[0048] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a four-week cycle comprising administering the compound for three weeks, followed by a one-week period without administration of the compound.

[0049] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered twice daily for two days per week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of eight weeks. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first and second days of each week.

[0050] In some embodiments, the cancer is selected from lung cancer, gastric cancer, liver cancer, colon cancer, kidney cancer, breast cancer, pancreatic cancer, pancreatic ductal adenocarcinoma (PDAC), juvenile myelomonocytic leukemia, neuroblastoma, melanoma, and acute myeloid leukemia. [Brief explanation of the drawing]

[0051] [Figure 1] Figure 1 shows data demonstrating that the compound of formula I and AMG510 synergistically bind to inhibit cell proliferation in NCI-H358 cells with the KRAS G12C mutation. [Figure 2A] Figure 2A shows plots of activity percentage versus inhibitor concentration (logM) in NCI-H358 cells treated with the compound of formula I alone and in combination with AMG510. IC50 data for NCI-H358 cells treated with the compound of formula I alone and in combination with AMG510 are shown in the table. [Figure 2B] Figure 2B shows a bar graph of CTG activity percentages, indicating that AMG510 (1 nM) alone did not reduce cell viability in NCI-H358 cells. [Figure 3] Figure 3 shows data demonstrating that the compound of formula I and AMG510 synergistically bind to inhibit cell proliferation in the KRAS G12C mutant NCI-H2122. [Figure 4A] Figure 4A shows plots of activity percentage versus inhibitor concentration (logM) in NCI-H2122 cells treated with the compound of formula I alone and in combination with various concentrations of AMG510. [Figure 4B] Figure 4B shows a bar graph of CTG activity percentages, indicating that AMG510 (1 nM) alone did not reduce cell viability in NCI-H2122 cells. [Figure 5] Figure 5 shows a matrix representation of HSA synergies and antagonists, demonstrating that the compound of formula I and adagrasib are synergistically combined to inhibit cell proliferation in KRAS G12C mutant NCI-H358 cells. [Figure 6A] Figure 6A shows a plot of activity percentage versus inhibitor concentration (logM) in NCI-H358 cells treated with the compound of formula I alone (black circles) and in combination with 1 nM adagrasib (black squares). [Figure 6B] Figure 6B shows a bar graph of CTG activity percentages, indicating that 1 nM adagrasib alone did not reduce cell viability in NCI-H358 cells. [Figure 7]Figure 7 shows a matrix representation of HSA synergistic and antagonistic effects, demonstrating that the compound of formula I and adagrasib are synergistically combined to inhibit cell proliferation in KRAS G12C mutant NCI-H2122 cells. [Figure 8] Figure 8 shows a matrix representation of HSA synergistic and antagonistic effects, demonstrating that the compound of formula I and adagrasib are synergistically combined to inhibit cell proliferation in KRAS G12C mutant KYSE-410 cells. [Figure 9A] Figure 9A shows plots of activity percentage versus inhibitor concentration (logM) in NCI-H2122 cells treated with adagrasib compound I alone (black circles, line 1) and in combination with 1 nM (black squares, line 2), 5 nM (black circles, line 3), or 10 nM (black squares, line 4). [Figure 9B] Figure 9B shows bar graphs of percentage CTG activity, indicating that adagrasib alone at 1 nM, 5 nM, or 10 nM concentrations did not reduce cell viability in NCI-H2122 cells. [Figure 10A] Figure 10A shows plots of tumor volume (mm3) versus treatment duration (days) for KRAS G12C mutant CRC022 PDX tumor xenograft models treated with vehicle (black circles, line 1), adagrasib alone (30 mg / kg QD, solid triangle, line 2), compound of formula I alone at 10 mg / kg / dose BID (black circles, line 3), compound of formula I at 30 mg / kg QD (black triangle, line 4), compound of formula I (10 mg / kg / dose BID) and adagrasib at 30 mg / kg QD (black circles, line 5), and compound of formula I (30 mg / kg QD) and adagrasib at 30 mg / kg QD (black triangle, line 6). [Figure 10B]Figure 10B shows a plot of tumor volume (mm3) versus treatment duration (days) of KRAS G12C mutation H2122 CDX tumor xenograft models treated with vehicle (black circles, line 1), adagrasib alone (30 mg / kg QD, solid triangle, line 2), compound of formula I alone (10 mg / kg / dose BID, black circles, line 3), compound of formula I alone at 30 mg / kg QD (black triangle, line 4), compound of formula I (10 mg / kg / dose BID) and adagrasib at 30 mg / kg QD (black circles, line 5), and compound of formula I (30 mg / kg QD) and adagrasib at 30 mg / kg QD (black triangle, line 6). [Figure 11] Figure 11 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model SW1573. [Figure 12] Figure 12 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model NCI-H358. [Figure 13] Figure 13 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant esophageal squamous cell carcinoma CDX model KYSE-410. [Figure 14] Figure 14 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model CO-04-0310. [Figure 15] Figure 15 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model CR2528. [Figure 16A] Figure 16A shows graphs of tumor volume over treatment periods for regimens of compound I alone (30 mg / kg QD), sotorasib (100 mg / kg QD) alone, and a combination of compound I (30 mg / kg QD) and sotorasib (100 mg / kg QD) in the KRAS G12C mutant NSCLC CDX model NCI-H2122. [Figure 16B] Figure 16B shows plots of tumor volume versus treatment duration (days) for KEAP1 mutant and KRAS G12C mutant NSCLC CDX model NCI-H2122 tumor xenograft models, treated with vehicle (black circles, line 1), sotorasib alone (100 mg / kg QD, black circles, line 2), compound of formula I alone (10 mg / kg / dose BID, black circles, line 3), and a combination of compound of formula I (10 mg / kg / dose BID) and sotorasib (100 mg / kg QD, black circles, line 4). [Figure 17] Figure 17 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model CRC022. Detailed description of the invention

[0052] I. General This embodiment is a method for treating a subject having cancer, wherein the subject is given a therapeutically effective amount of formula I:

[0053] [ka] The present invention provides a method comprising the step of administering a compound of formula I or a pharmaceutically acceptable salt thereof in combination with an inhibitor of KRAS having a G12C mutation. The following examples demonstrate the unexpected synergistic effects of the combination. The combination therapies disclosed herein using a compound of formula I or a pharmaceutically acceptable salt thereof can show superior results compared to alternative combinations of SHP2 inhibitors used in combination with an inhibitor of KRAS having a G12C mutation. Furthermore, the combination of the SHP2 inhibitor compound of formula I and the inhibitor of KRAS G12C provides a method that allows the use of lower doses of either agent used alone in monotherapy, which may help reduce potential side effects. In particular, the combination therapy may be effective in cancer cells expressing the G12C mutation. Thus, such treatment is suitable for the use of companion diagnostics to assist in the selection of an appropriate patient population. These and other advantages will be recognized by those skilled in the art.

[0054] Human KRAS G12C mutant tumors retained significant endogenous nucleotide cycling between their active (GTP-bound) and inactive (GDP-bound) states. KRAS G12C inhibitors (G12Ci) showed promising activity by binding to the inactive state of KRAS (GDP-bound) and preventing its reactivation via nucleotide exchange. Negative feedback activation of RTK and one of its downstream mediator proteins, SHP2, acted as a potential adaptive resistance mechanism. SHP2 is required for guanine nucleotide cycling, and its activity promoted growth in KRAS G12C tumors.

[0055] II. Definition Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art, as the embodiments described herein cover. Furthermore, any method or material similar to or equivalent to the methods or materials described herein may be used in the implementation of the embodiments herein. For the purposes of the embodiments disclosed herein, the following terms are defined:

[0056] As used herein, “A,” “an,” or “the” include not only embodiments having one element but also embodiments having multiple elements. For example, the singular forms “a,” “an,” and “the” include multiple referents unless explicitly indicated otherwise in the context. Thus, for example, a reference to “cells” includes multiple such cells, and a reference to “drugs” includes one or more drugs known to those skilled in the art.

[0057] "Pharmacologically acceptable excipients" refer to substances that assist in the administration and absorption of active agents to a subject. Useful pharmaceutical excipients in this embodiment include, but are not limited to, binders, fillers, disintegrants, lubricants, surfactants, coatings, sweeteners, flavorings, and colorants. Those skilled in the art will recognize that other pharmaceutical excipients may be useful in this embodiment.

[0058] "To treat," "to treat," and "treatment" refer to any sign of success in treating or improving a damage, condition, or disease, including any objective or subjective parameters such as reduction, remission, decrease or impairment of symptoms, making the condition or disease more tolerable to the patient, slowing the rate of degeneration or decline, reducing the debilitation of the final degenerative point, or improvement of the patient's physical or mental health. Treatment or improvement of symptoms may be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric examination, and / or psychiatric evaluation.

[0059] "Administration" refers to oral administration, topical contact as a suppository, parenteral administration, intravenous, intraperitoneal, intramuscular, intrafocal, intranasal or subcutaneous administration, intrathecal administration, or implantation of a sustained-release device to the subject, such as a mini osmotic pump. In the context of the combination therapies disclosed herein, administration may occur at separate times, simultaneously, or substantially simultaneously.

[0060] As used herein, “concurrent administration” or “combined administration” means administering one or more of the compositions described herein simultaneously with, immediately before, or immediately after, the administration of one or more additional therapies. The compounds provided herein may be administered alone or concurrently to a patient. Concurrent administration means simultaneous or consecutive administration of individual or combined compounds (multiple compounds). Concurrent administration means administration of compounds on the same day, in the same week, and / or within the same treatment schedule. Compounds may have different administration schedules, but if they are administered within the same treatment schedule, they may still be concurrently administered. For example, palbociclib may be administered once daily for three weeks within a four-week treatment schedule, and the compound of Formula I is concurrently administered with palbociclib if administered at any time within the four-week treatment schedule.

[0061] The “therapeutic dose” refers to the dose administered that produces a therapeutic effect. The exact dose depends on the purpose of the treatment and can be determined by those skilled in the art using known techniques (see, for example, Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992), Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999), Pickar, Dosage Calculations (1999), and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins), and all of the teachings therein, including but not limited to all methods, compounds, compositions, data, etc., for use with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety. In sensitized cells, the therapeutic dose may often be lower than the conventional therapeutic dose for unsensitized cells.

[0062] "Inhibit," "block," and "inhibitor" refer to compounds or methods that partially or completely block or interfere with a particular action or function.

[0063] "Cancer" refers to all types of cancer, neoplasms, or malignant tumors found in mammals (e.g., humans), including but not limited to leukemia, lymphoma, carcinoma, and sarcoma. Examples of cancers that can be treated with the compounds or methods provided herein include brain tumors, gliomas, glioblastomas, prostate cancer, colorectal cancer, pancreatic cancer, medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, head cancer, Hodgkin's disease, and non-Hodgkin lymphoma. Other examples of cancers that can be treated with the compounds or methods provided herein include cancers of the thyroid, endocrine system, brain, breast, cervix, colon, head and neck, liver, kidney, lung, ovaries, pancreas, rectum, stomach, and uterus. Further examples include thyroid cancer, bile duct cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma (PDAC), cutaneous melanoma, colon adenocarcinoma, rectal adenocarcinoma, gastric adenocarcinoma, esophageal cancer, head and neck squamous cell carcinoma, invasive breast cancer, lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung cancer, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyoma, primary thrombosis, primary macroglobulinemia, primary brain tumor, malignant pancreatic insulanoma, malignant carcinoid, bladder cancer, pre-malignant cutaneous lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical carcinoma, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid cancer, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, or prostate cancer.

[0064] The term "KRAS G12C inhibitor" generally refers to any inhibitor of KRAS having a G12C mutation. Such inhibitors include those known in the art that covalently bind to a 12-cysteine ​​residue, such as AMG510 (Amgen) and MRTX849 (Mirati). Other examples of KRAS G12C inhibitors are disclosed in the pending U.S. Provisional Patent Applications No. 63 / 082,221 (TRICYCLIC PYRIDONES AND PYRIMIDONES, filed September 23, 2020) and No. 63 / 116,146 (PYRROLIDINE-FUSED HETEROCYCLES, filed November 19, 2020) (these are respectively incorporated herein in their entirety by reference). In some embodiments, one or more inhibitors listed in this paragraph and elsewhere in this specification, and those in incorporated applications, may be specifically excluded from one or more embodiments described herein, including, but not limited to, any methods, kits, and compositions.

[0065] "Subject" refers to an organism that is suffering from or prone to a disease or illness that can be treated by administration of the pharmaceutical composition provided herein. Non-limiting examples include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, cows, deer, horses, and other non-mammals. In some embodiments, the patient is human.

[0066] III. Method of Administration In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as an oral composition.

[0067] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once or twice daily. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a continuous 28-day cycle.

[0068] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily in an amount of about 10 mg to about 140 mg.

[0069] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily in a three-week cycle, which includes administering the compound for two weeks, followed by a one-week period without administering the compound.

[0070] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily in a four-week cycle, which includes administering the compound for three weeks, followed by a one-week period without administration of the compound.

[0071] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of 6 weeks. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of 8 weeks.

[0072] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered three times a week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first, third, and fifth days of the week.

[0073] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered four times a week.

[0074] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in a three-week cycle, which includes administering the compound for two weeks, followed by a one-week period without administration of the compound.

[0075] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in a 4-week cycle, which includes administering the compound for 3 weeks, followed by a 1-week period without administration of the compound.

[0076] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily for two days a week. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of eight weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered on the first and second days of each week.

[0077] In some embodiments, the cancer is selected from lung cancer, gastric cancer, liver cancer, colon cancer, kidney cancer, breast cancer, pancreatic cancer, pancreatic ductal adenocarcinoma (PDAC), juvenile myelomonocytic leukemia, neuroblastoma, melanoma, and acute myeloid leukemia.

[0078] IV. Combination Methods In another aspect, the present disclosure relates to a method for treating a subject having cancer, wherein the subject is given a therapeutically effective amount of formula I:

[0079] [ka] A method is provided comprising the step of administering a compound or a pharmaceutically acceptable salt thereof in combination with an inhibitor of KRAS G12C. As disclosed herein, significant synergistic effects have been observed beyond those expected for such co-administration. Any suitable inhibitor, including any of those disclosed herein, may be used. Examples include, but are not limited to, AMG510 (sotorasib, LUMAKRAS®), MRTX849 (adagrasib), ARS-3248, GDC-6036, BI 1701963, tipifarnib, and BBP-454. In some embodiments, one or more inhibitors listed in this paragraph and elsewhere herein may be specifically excluded from embodiments described herein, including, but not limited to, any methods, kits, and compositions.

[0080] In some embodiments, the methods disclosed herein are suitable for treating any cancer having a KRAS G12C mutation. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is pancreatic ductal adenocarcinoma (PDAC). In some embodiments, the cancer is non-small cell lung cancer (NSCLC). In some embodiments, the cancer is cholangiocarcinoma. As will be understood by those skilled in the art, tumors can metastasize from the first or primary locus of the tumor to one or more other body tissues or sites. In particular, metastases to the central nervous system (i.e., secondary CNS tumors), and especially to the brain (i.e., brain metastases), are well documented for tumors and cancers, such as breast, lung, melanoma, kidney, and colorectal cancer. Thus, the methods disclosed herein can also be used for treating metastases to other organs (i.e., metastatic tumor growth).

[0081] In some embodiments, the method involves administering a third MAPK pathway inhibitor. While not theoretically bound, suppression of MAPK signaling in cancer cells may result in downregulation of PD-L1 expression, increasing the likelihood of cancer cells being detected by the immune system. Such third MAPK pathway inhibitors may be based on other mutations in proteins in the MAPK pathway. In some embodiments, any MAPK pathway inhibitor can be used, including those targeting KRAS, NRAS, HRAS, PDGFRA, PDGFRB, MET, FGFR, ALK, ROS1, TRKA, TRKB, TRKC, EGFR, IGF1R, GRB2, SOS, ARAF, BRAF, RAFI, MEK1, MEK2, c-Myc, CDK4, CDK6, CDK2, ERK1, and ERK2. Exemplary MAPK pathway inhibitors include, but are not limited to, afatinib, osimertinib, erlotinib, gefitinib, lapatinib, neratinib, dacomitinib, vandetanib, cetuximab, panitumumab, nimotuzumab, necitumumab, trametinib, binimetinib, cobimetinib, selumetinib, urixertinib, LTT462, and LY3214996. In some embodiments, but are not limited, one or more of the above-listed inhibitors, including any methods, kits, and compositions, may be specifically excluded from the embodiments described herein.

[0082] The methods disclosed herein can be combined with other chemotherapeutic agents. Examples of such agents can be found in Cancer Principles and Practice of Oncology by VTDevita and S. Hellman (editors), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers, which is incorporated herein by reference in its entirety, including, but not limited to, all methods, compounds, compositions, data, etc., for use in any of the embodiments and disclosures herein. Those skilled in the art will be able to identify which combinations of agents are useful based on the specific characteristics of the drugs and the diseases involved.

[0083] In some embodiments, the method may include the simultaneous administration of at least one cytotoxic agent. The term “cytotoxic agent,” as used herein, refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., At211, 1131, 1125, Y90, Rel86, Re188, Sm153, Bi212, P32, Pb212, and radioisotopes of Lu), chemotherapeutic agents, growth inhibitors, enzymes such as nucleases and their fragments, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin, including their fragments and / or variants.

[0084] Examples of cytotoxic agents can be selected from antimicrotubule agents, platinum coordination complexes, alkylating agents, antibiotics, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signaling pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, apoptosis promoters, LDH-A inhibitors; fatty acid biosynthesis inhibitors; cell cycle signaling inhibitors; HDAC inhibitors; proteasome inhibitors; and cancer metabolism inhibitors.

[0085] Chemotherapy agents include chemical compounds useful for treating cancer. Examples of chemotherapeutic agents include erlotinib (TARCEVA®, Genentech / OSI Pharm.), bortezomib (VELCADE®, Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX®, AstraZeneca), sunitinib (SUTENT®, Pfizer / Sugen), and letrozole (FEMARA®, Novartis). Imatinib mesylate (GLEEVEC®, Novartis), financiate (VATALANIB®, Novartis), oxaliplatin (ELOXATIN®, Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib (TYKERB®, GSK572016, Glaxo Smith Kline), ronafamib (SCH66336), sorafenib (NEXAVAR®, Bayer) Labs), gefitinib (IRESSA®, AstraZeneca), AG1478, alkylating agents, e.g., thiotepa and CYTOXAN® cyclophosphamide; alkyl sulfonates, e.g., busulfan, iprosulfan, and piposulfan; aziridines such as benzodopa, carboquan, metsuredopa, and uredopa; altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, etc. Ethyleneimines and methylamelamine, including trimethylomelamine; acetogenins (especially bratacin and bratacinone); camptothecin (including topotecan and irinotecan); briostatin; calcistatin; CC1065 (including its adzeresin, karzeresin and bizeresin synthetic analogs); cryptophycin (especially cryptophycin 1 and cryptophycin 8); corticosteroids (including prednisone and prednisolone); cyproterone acetate;5-α-reductase (including finasteride and dutasteride); vorinostat, romidepsin, panobinostat, valproic acid, mosetinostat, dorastatin; aldesleukin, talc duocalmycin (including synthetic analogs KW-2189 and CB1-TM1); eluterobin; cratistatin; sarcoditine; spongstatin; chlorambucil, chromafazine, chlorophosphamide, estramustine, ifosfamide, mechlore Nitrogen mustards such as tamin, mechloretamine oxide hydrochloride, melphalan, nobemubikin, fenestrine, prednimustine, trophosphamide, and uracil mustard; nitrosoureas, e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, e.g., engine antibiotics (e.g., calicheamicin, in particular calicheamicin γ11 and calicheamicin ω1) (Angew Chem. Int. Ed. Engl. 1994) 33:183-186); Dynemycin containing dynemycin A; Bisphosphonates, e.g., clodronate; Esperamycin; Similarly, neocardinostatin chromophores and related pigment proteins (endiin antibiotic chromophores), actinomycin, actinomycin, ausramycin, azaserin, bleomycin, kactinomycin, carabicin, aminomycin, cardinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), mo (Ruhorino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin such as mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, porphyromycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, yubenimex, dinostatin, zolubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU);Folic acid analogs, e.g., denopterin, methotrexate, pteropterin, trimethrexate; purine analogs, e.g., fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, e.g., acitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenal drugs such as aminoglutethimide, mitotane, trilostane; folic acid supplements, e.g., floric acid; acegraton; aldofsphamide glycosides; aminolevulinic acid; enyluracil; amsacrin; bestrabusil; bisanthren; edatraxate; defomin; demecolcine; diazicon; elf Ornithine; eriptinium acetate; epotilone; etoglucide; gallium nitrate; hydroxyurea; lentinan; ronidynin; mayansinoids, e.g., maytansine and anthamitosine; mitogwazone; mitoxantrone; mopidamnol; nitroserine; pentostatin; phenamet; pirarubicin; rosoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; PK (registered trademark) polysaccharide complex (JHS Natural) Products, Eugene, Oreg.); Lazoxane; Rhizoxin; Schizofuran; Spirogermanium; Tenuazonic Acid; Triadicone; 2,2',2”-Trichlorotriethylamine; Trichothecenes (especially T-2 toxin, Veraculine A, Loridine A, and Angidin); Urethanes; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacitosine; Arabinoside ("Ara-C"); Cyclophosphamide; Thiotepa; Taxoids, e.g., TAXOL (Paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ.), ABRAXANE® (Cremophor-free), Albumin-Modified Nanoparticle Formulations of Paclitaxel (American Pharmaceutical Partners, Schaumberg, IL), and TAXOTERE® (Docetaxel; Doxetaxel;Examples include Sanofi-Aventis; chlorambucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE® (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids, e.g., retinoic acid; and any pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0086] Chemotherapy agents include (i) antihormone agents that act to modulate or inhibit the hormonal effects on tumors, such as anti-estrogen and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (NOLVADEX®; including tamoxifen citrate), raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, and FARESTON® (toremifene citrate); and (ii) es in the adrenal gland. Aromatase inhibitors that inhibit the enzyme aromatase, which regulates trogen production, such as 4(5)-imidazole, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestany, fadrozol, RIVISOR® (borozol), FEMARA® (letrozole; Novartis), and ARIMIIDEX® (anastrozole; AstraZeneca); (iii) flutamide, nilutamide, bicalutamide, leuproli (iv) Antiandrogens such as buserelin, trypterine, medroxyprogesterone acetate, diethylstilbestrol, Premarin, fluoxymesterone, all trans-retinic acid, fenretinide, and troxacitabine (1,3-dioxolane nucleoside cytosine analog); (iv) Protein kinase inhibitors; (v) Lipid kinase inhibitors; (vi) Antisense oligonucleotides, in particular those that inhibit gene expression in signaling pathways involved in abnormal cell proliferation, e.g., PKC-α, Ralf, etc. (vii) Ribozymes such as VEGF expression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii) Vaccines such as gene therapy vaccines, e.g., ALLOVECTIN®, LEUVECTIN®, and VAXID®; PROLEUKIN®, rIL-2; Topoisomerase 1 inhibitors such as LURTOTECAN®; ABARELIX® rmRH; and (ix) pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0087] Chemotherapy agents include antibodies such as alemtuzumab (Campath) and bevacizumab (AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); panitumumab (VECTIBIX®, Amgen); rituximab (RITUXAN®, Genentech / Biogen Idee); pertuzumab (OMNITARG®, 2C4, Genentech); trastuzumab (HERCEPTIN®, Genentech); tositumomab (Bexxar, Corixia); and the antibody-drug conjugate gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Further humanized monoclonal antibodies that have therapeutic potential as drugs in combination with the compounds of the present invention include ipilimumab, rabetuzumab, lintuzumab, matuzumab, mepolizumab, motabizumab, motobizumab, natalizumab, nimotuzumab, norovizumab, numabizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pefstuzumab, pectuzumab, pexelizumab, and larivizumab. These include ranibizumab, recibizumab, recilizumab, recibizumab, loberizumab, luprizumab, cibrotuzumab, ciprizumab, sontuzumab, tacutuzumab tetraxetan, tadocizumab, talizumab, tefivazumab, tocilizumab, tralizumab, tucotsuzumab cermoloykin, tuccituzumab, umabizumab, urtoxazumab, ustekinumab, bicilizumab, and anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), which is a recombinant exclusive human full-length IgG1λ antibody genetically modified to recognize the interleukin-12 p40 protein.

[0088] Chemotherapy agents also include "EGFR inhibitors," which refer to compounds that bind to EGFR or its variant forms, or otherwise directly interact with it, preventing or reducing its signaling activity; these are also called "EGFR antagonists." Examples of such drugs include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRLHB 8506), MAb 455 (ATCC CRLHB 8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al.), as well as chimeric 225 (C225 or cetuximab; ERBUTIX®) and remodeled human 225 (H225) (WO96 / 40210, Imclone Systems). See Inc. for variants thereof; IMC-11F8 (Imclone), a fully human EGFR target antibody; antibodies that bind to type II variant EGFR (U.S. Patent No. 5,212,290); humanized and chimeric antibodies that bind to EGFR as described in U.S. Patent No. 5,891,996; and human antibodies that bind to EGFR, e.g., ABX-EGF or panitumumab (see WO98 / 50433, Abgenix / Amgen); EMD55900 (Stragliotto et al. Eur.J. Cancer). Examples include 32A:636-640(1996)); EMD7200 (matsuzumab), a humanized EGFR antibody that competes with both EGF and TGFα for EGFR binding (EMD / Merck); the human EGFR antibody, HuMax-EGFR (GenMab), also known as E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3, a fully human antibody described in US Patent No. 6,235,883; MDX-447 (Medarex Inc); and mAb806 or humanized mAb806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)).Anti-EGFR antibodies can be conjugated with cytotoxic agents, thus generating immunoconjugates (see, for example, EP659,439A2, Merck Patent GmbH). EGFR antagonists include U.S. Patent Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, 5,866, This includes small molecules such as compounds described in publications 572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008, and 5,747,498, as well as PCT publications WO98 / 14451, W098 / 50038, W099 / 09016, and WO99 / 24037.Specific small molecule EGFR antagonists include OSI-774 (CP-358774), erlotinib, TARCEVA® Genentech / OSI Pharmaceuticals); PD183805 (CI 1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca; ZM 105180((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-182(N8-(3-chloro-4-fluorophenyl)-N2-(l-methyl-piperidine-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166((R)-4-[4-[(l-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidine-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(l-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butinamide); EKB-569(N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide)(Wyeth); AG1478(Pfizer); AG1571(SU5271;Pfizer); Dual EGFR / HER2 tyrosine kinase inhibitors, such as lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine]). Each of the above references is incorporated herein by reference in its entirety, with respect to all of its teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein.

[0089] Chemotherapy agents also include "tyrosine kinase inhibitors," including the EGFR-targeted drugs mentioned in the previous paragraph; small molecule HER2 tyrosine kinase inhibitors such as TAK165 available from Takeda; CP-724, 714, oral selective inhibitors of ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual-HER inhibitors such as EKB-569 (available from Wyeth), which inhibits both HER2 and EGFR overexpressing cells and preferentially binds to EGFR but; lapatinib (GSK572016; available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033; Pharmacia); and ISIS, which inhibits Raf-1 signaling. Antisense agents available from Pharmaceuticals include Raf-1 inhibitors such as ISIS-5132; non-HER-targeted TK inhibitors, e.g., imatinib mesylate (GLEEVEC®, available from Glaxo SmithKline); multi-targeted tyrosine kinase inhibitors, e.g., sunitinib (SUTENT®, available from Pfizer); VEGF receptor tyrosine kinase inhibitors such as batalanib (PTK787 / ZK222584, available from Novartis / Schering); MAPK extracellular regulatory kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines, e.g., PD153035,4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyridopyrimidines; pyrrolopyrimidines, e.g., CGP59326, CGP60261, etc. CGP62706; pyrazolopyrimidine, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tilphostin containing the nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those that bind to nucleic acids encoding HER); quinoxaline (US Patent No. 5,804,396); triphostin (US Patent No. 5,804,396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering);pan-HER inhibitors, e.g., CI-1033 (Pfizer); Affinitac (ISIS 3521; ISIS / Lilly); Imatinib mesylate (GLEEVEC®); PKI166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering); INC-1C11 (Imclone); Rapamycin (Sirolimus, RAPAMUNE®); or U.S. Patent No. 5,804,396; WO1999 / 09016 (American Cyanamid); WO1998 / 43960 (American Cyanamid); WO1997 / 38983 (Warner As described in any of the following patent publications: Lambert); WO1999 / 06378 (Warner Lambert); WO1999 / 06396 (Warner Lambert); WO1996 / 30347 (Pfizer, Inc); WO1996 / 33978 (Zeneca); WO1996 / 3397 (Zeneca); and WO1996 / 33980 (Zeneca). Each of the above references is incorporated herein by reference in its entirety, with respect to all teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein.

[0090] Chemotherapy agents include dexamethasone, interferon, colchicine, methoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostin, arsenic trioxide, asparaginase, BCG Live, bevacuzimab, besarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexazoxane, epoetin alfa, erotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, and interferon alfa. This also includes Phalen, cyclosporine, nandrolone, nelarabine, nofetumomab, oprelbequine, parifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexeddisodium, plicamycin, porfimer sodium, quinacrine, rasburicase, salglamostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, barulurubicin, zoledronate, and zoledronic acid, as well as pharmaceutically acceptable salts thereof.

[0091] Chemotherapy agents include hydrocortisone, hydrocortisone acetate, cortisone acetate, thixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortone, and hydrocortisone-17-butyrate. Hydrocortisone-17-valerate, acromethasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, fluocortone caproate, fluocortone pivalate, and flupredniden acetate; phenylalanine-glutamine-glycine (FEG) and its D-isomer (FEG) (IMULAN Immunoselective anti-inflammatory peptides (ImSAIDs) such as BioTherapeutics, LLC; antirheumatic drugs, e.g., azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salt, hydroxychloroquine, leflunomidocycline, sulfasalazine; tumor necrosis factor alpha (TNFa) blockers, e.g., etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), golimumab (Simponi); interleukin-1 (IL-1) blockers, e.g., anakinra (Kineret); T-cell costimulatory blockers, e.g., abatacept (Ore) NCIs); interleukin-6 (IL-6) blockers, e.g., tocilizumab (ACTEMERA®); interleukin-13 (IL-13) blockers, e.g., lebrikizumab; interferon-alpha (IFN) blockers, e.g., lontalizumab; beta-7 integrin blockers such as rhuMAb beta-7; IgE pathway blockers such as anti-M1 prime; secretory homotrimer LTa3 and membrane-bound heterotrimer LTa / p2 blockers, e.g., antilymphotoxin α (LTa); radioactive isotopes (e.g., At211, 1131, 1125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, and radioactive isotopes of Lu);Other research agents such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3, olfamesyltransferase inhibitors (L-739749, L-744832); polyphenols, e.g., quercetin, resveratrol, piceatannol, epigallocatechin gallate, theaflavins, flavanols, procyanidins, betulinic acid, and their derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapacone; apachol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin; podophyllotoxin; tegafur (UFTORAL®); bexarotene (TARGRETIN®); clodronate (e.g., BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), pamidronate Bisphosphonates such as (AREDIA®), tildrone (SKELID®), or risedronate (ACTONEL®); and epidermal growth factor receptor (EGF-R); vaccines such as THERATOPE® vaccine; perifosin, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); CCI-779; tipifarnib (R11577); olafenib, ABT510; oblimersen sodium (GENASENSE) This includes Bcl-2 inhibitors such as (registered trademark)); pixantrone; farnesyltransferase inhibitors such as ronafamib (SCH6636, SARASARTM); and any pharmaceutically acceptable salts, acids, or derivatives of any of the above; two or more combinations of the above, such as CHOP, an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone; and FOLFOX, an abbreviation for treatment regimens with oxaliplatin (ELOXATIN®) in combination with 5-FU and leucovorin.

[0092] Chemotherapy agents also include nonsteroidal anti-inflammatory drugs (NSAIDs) that have analgesic, antipyretic, and anti-inflammatory effects. NSAIDs include non-selective inhibitors of the enzyme cyclooxygenase. Specific examples of NSAIDs include propionic acid derivatives such as aspirin, ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, and naproxen; acetic acid derivatives such as indomethacin, sulindac, etodolac, and diclofenac; enolic acid derivatives such as piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, and isoxicam; fenamic acid derivatives such as mefenamic acid, meclofenamic acid, flufenamic acid, and tolfenamic acid; and COX-2 inhibitors such as celecoxib, etoricoxib, lumiracoxib, parecoxib, lofecoxib, and vardecoxib. NSAIDs may be indicated for the relief of symptoms of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthritis, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headaches and migraines, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, ileus, and colitis.

[0093] In certain embodiments, chemotherapeutic agents include, but are not limited to, doxorubicin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, interferon, platinum derivatives, taxanes (e.g., paclitaxel, docetaxel), vinca alkaloids (e.g., vinblastine), anthracyclines (e.g., doxorubicin), epipodophilotoxin (e.g., etoposide), cisplatin, mTOR inhibitors (e.g., rapamycin), methotrexate, actinomycin D, drastatin 10, colchicine, trimethrexate, methoprine, cyclosporine, daunorubicin, teniposide, amphotericin, alkylating agents (e.g., chlorambucil), 5-fluorouracil, camptothecin, cisplatin, metronidazole, and imatinib mesylate. In other embodiments, the compounds disclosed herein are administered in combination with a biological agent such as bevacizumab or panitumumab.

[0094] In certain embodiments, the compounds disclosed herein, or pharmaceutically acceptable compositions thereof, include avalerix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG Live, bevacuzimab, fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, carsterone, capecitabine, camptothecin, carboplatin, carmustine, cetuximab, chlorambucil, cladribine, and clofalabi. Cyclophosphamide, Cytarabine, Dactinomycin, Darbepoetin alfa, Daunorubicin, Denileukin, Dexazoxane, Docetaxel, Doxorubicin (neutral), Doxorubicin hydrochloride, Dromostanolone propionate, Epirubicin, Epoetin alfa, Erotinib, Estramustine, Etoposide phosphate, Etoposide, Exemestane, Filgrastim, Furoxuridine, Fludarabine, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab, Goserelin acetate, Histrelin acetate, Hydroxyurea, Ibri Tumomab, idarubicin, ifosfamide, imatinib mesylate, interferon alpha-2a, interferon alpha-2b, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamizole, lomustine, megestrol acetate, melphalan, mercaptopurine, 6-MP, mesna, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone, nelarabine, nofetumomab, oprelbequin, oxaliplatin, paclitaxel, parifen Lumin, Pamidronate, Pegademase, Pegaspargase, Pegfilgrastim, Pemetrexeddisodium, Pentostatin, Pipobroman, Plicamycin, Porfimer sodium, Procarbazine, Quinacrine, Rasburicase, Rituximab, Salglamostim, Sorafenib, Streptozocin, Sunitinib maleate, Talc, Tamoxifen, Temozolomide, Teniposide, VM-26, Testolactone, Thioguanine, 6-TG, Thiotepa, Topotecan, Toremifene, Tositumomab, Trastuzumab, Tretinoin, ATRA,It is administered in combination with one or more antiproliferative agents or chemotherapeutic agents selected from uracil mustard, barrubicin, vinblastine, vincristine, vinorelbine, zoledronate, or zoledronic acid.

[0095] In some embodiments, the dosage of the compound of formula I may be any appropriate amount to treat cancer. For example, the dosage may be a daily dose between 1 mg and 500 mg by weight. As a further example, the daily dose may be in the range of about 20 mg to 400 mg (or any partial range or partial value (including endpoints) in that range). In some embodiments, the dosage range of the compound of formula I may be 10 mg to 300 mg. In some embodiments, the dosage range of the compound of formula I may be 10 mg to 100 mg. In some embodiments, the dosage range of the compound of formula I may be 5 mg to 50 mg. The daily dose can be achieved by administering a single dose (e.g., QD) or by providing a total daily dose through multiple doses over a day (e.g., BID, TID, QID, etc.). In some embodiments, the dosage of the KRAS inhibitor may be any appropriate amount. For example, it may be an amount in the range of 1 mg to 1,000 mg per day (or any partial range or partial value (including endpoints) in that range). Dosage of KRAS inhibitors may be the same as or less than the approved dosage for any given KRAS inhibitor, and may depend on the given indication. For example, AMG510 may be administered once daily in the range of 500 mg to 1,000 mg. For example, MRTX849 may be administered once daily in the range of 500 mg to 1,200 mg. It will be understood that each of the listed ranges above may include any subrange or subpoint within it, including the endpoint. It will be understood that each of the listed ranges above may include any partial range or partial point within it, including the endpoint. The general dose range for adults is generally 5 mg to 2 g / day. Tablets or other forms of presentation offered in individual units may conveniently contain units containing one or more compounds in amounts effective as such doses or multiples thereof, e.g., 5 mg to 500 mg, usually about 10 mg to 200 mg. The amount of active ingredient that can be combined with a carrier material to produce a single-dose formulation varies depending on the host being treated and the specific mode of administration. In some embodiments, administration is orally.

[0096] In some embodiments, a method is provided for treating lung cancer or esophageal cancer in a subject, comprising the step of orally administering a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof to the subject in combination with AMG510. In some embodiments, the compound of formula I is administered once or twice daily. In some embodiments, AMG510 is administered once or twice daily. The drugs may be administered concurrently, for example, as described herein.

[0097] In some embodiments, a method is provided for treating lung cancer, colorectal cancer, esophageal cancer, or breast cancer in a subject, comprising the step of orally administering a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof to the subject in combination with adagrasib. In some embodiments, the compound of formula I is administered once or twice daily. In some embodiments, adagrasib is administered once or twice daily. The drugs may be administered concurrently, for example, as described herein.

[0098] In some embodiments, the subject is human. In some embodiments, the subject is a non-human mammal such as a primate, rodent, dog, cat, or other small animal.

[0099] composition The compounds of formula I disclosed herein may exist as salts. This embodiment includes such salts that may be pharmaceutically acceptable. Examples of applicable salt forms include salts with hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate (e.g., (+)-tartrate, (-)-tartrate) or racemic mixtures thereof, succinate, benzoate, and amino acids such as glutamic acid. These salts can be prepared by methods known to those skilled in the art. Also included are base addition salts such as sodium, potassium, calcium, and ammonium, organic amino or magnesium salts, or similar salts. If the compounds of this embodiment contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogenic acid, phosphoric acid, monohydrogenic acid, dihydrogenic acid, sulfuric acid, monohydrogenic acid, hydroiodic acid, or phosphorous acid, and salts derived from organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids such as alginates and salts of organic acids such as glucuronic acid or galacturonic acid are also included. Certain compounds in this embodiment contain both basic and acidic functional groups that enable the conversion of the compound into either a base addition salt or an acid addition salt.

[0100] Other salts include acid or base salts of the compounds used in the methods of this embodiment. Examples of pharmaceutically acceptable salts include mineral acid salts (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid), organic acid salts (e.g., acetic acid, propionic acid, glutamic acid, citrate), and quaternary ammonium salts (e.g., methyl iodide, ethyl iodide). It is understood that pharmaceutically acceptable salts are nontoxic. Further information on suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa, 1985, which incorporates, by reference, all of its teachings, including but not limited to all methods, compounds, compositions, data, etc., for use with any of the embodiments and disclosures herein.

[0101] Pharmaceutically acceptable salts include salts of the active compound prepared with relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein. If the compounds of this embodiment contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts. If the compounds of this embodiment contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogenic acid, phosphoric acid, monohydrogenic acid, dihydrogenic acid, sulfuric acid, monohydrogenic acid, hydroiodic acid, and phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. This also includes salts of amino acids such as alginates, and salts of organic acids such as glucuronic acid or galacturonic acid (see, for example, Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19), and all teachings of which, without limitation, include all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety. Certain compounds in these embodiments contain both basic and acidic functional groups that enable the conversion of the compound into either a base-addition salt or an acid-addition salt.

[0102] The neutral form of the compound is preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0103] Certain compounds of this embodiment can exist in solvated forms, including non-solvated and hydrated forms. Generally, the solvated forms are equivalent to the non-solvated forms and are included within the scope of this embodiment. Certain compounds of this embodiment can exist in multiple crystalline or amorphous forms. Generally, all physical forms are equivalent to the uses intended by this embodiment and are intended to be within the scope of this embodiment.

[0104] Certain compounds of this embodiment have an asymmetric carbon atom (optical center) or a double bond and can be defined in terms of absolute stereochemistry as (R)-, (S)-, (D)-, or (L)- for amino acids. Enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomer forms, and individual isomers are included within the scope of this embodiment. The compounds of this embodiment do not include those known in the art to be too unstable to synthesize and / or isolate. This embodiment means that compounds are included in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or divided using conventional techniques.

[0105] Unless otherwise specified, the compounds of this embodiment may also contain non-natural proportion atomic isotopes in one or more of the atoms constituting such compounds. For example, the compounds of this embodiment may contain radioactive or stable isotopes, such as deuterium ( 2 H), tritium ( 3 H), Iodine-125( 125 I), Fluorine-18( 18 F), Nitrogen-15( 15 N), oxygen-17( 17 O), oxygen-18( 18O), carbon-13( 13 C), or carbon-14( 14 C) and the like. All isotope variations of the compounds of the embodiments of the present invention are included within the scope of the embodiments of the present invention, whether radioactive or not.

[0106] In addition to the salt form, this embodiment provides compounds in prodrug form. The prodrugs of the compounds described herein are compounds that readily undergo chemical change under physiological conditions to provide the compounds of the embodiments of the present invention. Further, the prodrug can be converted to the compound of the embodiment of the present invention by chemical or biochemical methods in an ex vivo environment. For example, the prodrug can be slowly converted to the compound of the embodiment of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0107] In some embodiments, pharmaceutical compositions comprising a compound of formula I and a pharmaceutically acceptable excipient are provided. In some embodiments, the pharmaceutical composition is configured as an oral tablet.

[0108] The compounds of the embodiments of the present invention can be prepared and administered in a wide variety of oral, parenteral, and topical dosage forms. Oral formulations include tablets, pills, powders, sugar-coated tablets, capsules, liquids, lozenges, gels, syrups, slurries, and suspensions suitable for patient intake. The compounds of the embodiments can also be administered by injection, i.e., intravenously, intramuscularly, intradermally, subcutaneously, intradurally, or intraperitoneally. Furthermore, the compounds described herein can be administered by inhalation, for example, intranasally. In addition, the compounds of the embodiments of the present invention can be administered transdermally. The compounds of formula I disclosed herein may also be administered by intraocular, intravaginal, and intrarectal routes, including suppositories, inhalants, powders, and aerosol formulations (for example, see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111, 1995), and all teachings, including but not limited to all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety. Accordingly, these embodiments also provide pharmaceutical compositions comprising one or more pharmaceutically acceptable carriers and / or excipients and either a compound of formula I or a pharmaceutically acceptable salt of a compound of formula I.

[0109] For preparing pharmaceutical compositions from the compounds of embodiments of the present invention, pharmaceutically acceptable carriers may be either solid or liquid. Preparations in solid form include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. The solid carrier may be one or more substances that also act as diluents, flavoring agents, surfactants, binders, preservatives, tablet disintegrants, or encapsulating materials. Further details regarding the techniques for formulation and administration are described in detail in scientific literature and patent documents, for example, the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's"), which, for use in conjunction with any of the embodiments and disclosures herein, incorporates herein by reference all of its teachings, including but not limited to all methods, compounds, compositions, data, etc.

[0110] In the case of a powder, the carrier is a finely ground solid that is a mixture with the finely ground active ingredient. In the case of a tablet, the active ingredient is mixed in appropriate proportions with a carrier having the required binding properties and additional excipients as needed, and then compressed into the desired shape and size.

[0111] Powders, capsules, and tablets preferably contain 5% or 10% to 70% of the active compound. Suitable carriers include magnesium carbonate, magnesium stearate, talc, sugars, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low-melting-point wax, and cocoa butter. The term “preparation” is intended to include formulations of the active compound having an encapsulating material as a carrier, in which the active ingredient, with or without other excipients, is surrounded by a carrier, and thus provides a capsule associated with the carrier. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0112] Suitable solid excipients include, but are not limited to, sugars containing lactose, sucrose, mannitol, or sorbitol; starches derived from corn, wheat, rice, potato, or other plants; celluloses such as methylcellulose, hydroxypropyl methylcellulose, and sodium carboxymethylcellulose; and gums containing gum arabic and gum tragacanth; in addition, carbohydrate or protein fillers containing proteins such as gelatin and collagen. If desired, disintegrants or solubilizers such as cross-linked polyvinylpyrrolidone, agar, alginic acid, or salts thereof such as sodium alginate may be added.

[0113] The sugar-coated tablet core is provided with suitable coatings such as concentrated sugar solutions that may also contain gum arabic, talc, polyvinylpyrrolidone, Carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablet or sugar-coated tablet coating for product identification or to characterize the amount of the active compound (i.e., dosage). The pharmaceutical preparations disclosed herein may also be used orally using, for example, push-fit capsules made of gelatin, and soft-seal capsules made of gelatin and a coating (e.g., glycerol or sorbitol). Push-fit capsules may contain the compound of formula I mixed with a filler or binder such as lactose or starch, a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the compound of formula I may be dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin, or liquid polyethylene glycol, with or without a stabilizer.

[0114] Liquid preparations include solutions, suspensions, and emulsions, such as water or water / propylene glycol solutions. For parenteral injection, liquid preparations may be formulated in solution in an aqueous polyethylene glycol solution.

[0115] An aqueous solution suitable for oral use can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavorings, stabilizers, and thickeners as desired. Aqueous suspensions suitable for oral use can be prepared by dispersing the micronized active ingredient in water with a viscous material such as natural or synthetic rubber, resin, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth rubber, and acacia rubber, and a dispersant or wetting agent such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of alkylene oxide and fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide and long-chain aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensation product of ethylene oxide and a partial ester derived from fatty acid and hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensation product of ethylene oxide and a partial ester derived from fatty acid and hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavoring agents, and one or more sweeteners, such as sucrose, aspartame, or saccharin. The formulation can be adjusted in terms of volumetric osmolality.

[0116] This also includes solid preparations intended to be converted into liquid preparations for oral administration immediately before use. Such liquid preparations include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active ingredient, colorants, flavorings, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.

[0117] The oily suspension can be formulated by suspending the compound of Formula I in a vegetable oil such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin; or in a mixture thereof. The oily suspension may contain a thickener such as beeswax, solid paraffin, or cetyl alcohol. Sweeteners such as glycerol, sorbitol, or sucrose can be added to provide an oral preparation with a pleasant mouthfeel. These formulations can be preserved by adding antioxidants such as ascorbic acid. For an example of an injectable oil vehicle, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997, which discloses any embodiment of any of the methods, compounds, compositions, and data, without being particularly limited, encompassing all of the invention. The pharmaceutical formulations disclosed herein may also be in the form of an oil-in-water emulsion. The oil phase may be the vegetable oil or mineral oil described above, or a mixture thereof. Suitable emulsifiers include naturally occurring rubbers, such as acacia rubber and tragacanth rubber; naturally occurring phosphatides, such as soy lecithin; esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate; and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. Emulsions may also contain sweeteners and flavorings, as in the case of syrup and elixir formulations. Such formulations may also contain lubricants, preservatives, or colorants.

[0118] Pharmaceutical formulations of the compounds of formula I disclosed herein can be provided as salts and can be formed with bases, i.e., cationic salts, such as alkali and alkaline earth metal salts, such as sodium, lithium, potassium, calcium, magnesium, and ammonium salts, such as ammonium, trimethylammonium, diethylammonium, and tris-(hydroxymethyl)-methylammonium salts.

[0119] Pharmaceutical preparations are preferably in unit dosage forms. In such forms, the preparation is subdivided into unit doses containing an appropriate amount of the active ingredient. A unit dosage form may be a packaged preparation, and the packaging contains separate amounts of the preparation, such as packaged tablets, capsules, and powders in vials or ampoules. Alternatively, a unit dosage form may be a capsule, tablet, cachet, or lozenge itself, or an appropriate number of any of these in packaged form.

[0120] The amount of the active ingredient in a unit dose preparation may vary or be adjusted from 0.1 mg to 10,000 mg, more typically from 1.0 mg to 1,000 mg, and most typically from 10 mg to 500 mg, depending on the specific use and the potency of the active ingredient. The composition may also contain other suitable therapeutic agents if desired.

[0121] The administration regimen also takes into account pharmacokinetic parameters well known in the art, namely absorption rate, bioavailability, metabolism, clearance, etc. (for example, all teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety: Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci. 84:1144-1146; Rohatagi (1995) Pharmazie See 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24:103-108; the latest Remington's, supra). State-of-the-art technology enables clinicians to determine the dosage regimen for each individual patient, GR and / or MR modulator, and disease or illness being treated.

[0122] A single or multiple dose of a compound formulation of formula I can be administered according to the patient's needs and acceptable dosage and frequency. The formulation should provide a sufficient amount of the active agent to effectively treat the disease condition. Accordingly, in one embodiment, a pharmaceutical formulation for oral administration of a compound of formula I is a daily dose of about 0.5 to about 30 mg / kg body weight / day, including all partial ranges and partial values ​​therein, including the endpoint. In an alternative embodiment, the dose is about 1 mg to about 20 mg / kg body weight / patient / day. In particular, lower doses can be used when the drug is administered into an anatomically occluded site such as the cerebrospinal fluid (CSF) space, into the bloodstream, into a body cavity, or into the lumen of an organ, in contrast to oral administration. Substantially higher doses can be used for topical administration. Practical methods for preparing formulations containing a compound of formula I for parenteral administration are known or evident to those skilled in the art and are described in detail in publications such as Remington's, supra. See also Nieman, In “Receptor Mediated Antisteroid Action,” Agarwal, et al., eds., De Gruyter, New York (1987), which is incorporated herein by reference in its entirety for all teachings, including but not limited to all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein.

[0123] In some embodiments, co-administration involves administering one active agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours (or any sub-range of time or sub-value of time within 24 hours) of the second active agent. Co-administration involves administering two active agents simultaneously, nearly simultaneously (e.g., within approximately 1, 5, 10, 15, 20, or 30 minutes of each other (or, e.g., any sub-range of time or sub-value of time from 0 to 30 minutes)), or sequentially in any order. In some embodiments, co-administration can be achieved by co-formulation, i.e., by preparing a single pharmaceutical composition containing both active agents. In some embodiments, the active agents can be formulated separately. In some embodiments, the activators and / or adjuvants can be linked or conjugated to each other. At least one dose of the drugs can be administered, for example, simultaneously. At least one dose of the drugs can be administered, for example, within a few minutes or less than an hour of each other. At least one dose of the drug can be administered, for example, at different times, but on the same day or on different days.

[0124] A pharmaceutical composition comprising a compound of formula I disclosed herein may, after being formulated in one or more acceptable carriers, be placed in a suitable container and labeled for treatment of the indicated conditions. Such labeling for the administration of a compound of formula I may include, for example, instructions regarding the amount, frequency, and method of administration.

[0125] Administration of pharmaceuticals The administration regimens of the compounds described herein will naturally vary depending on known factors such as the recipient's species, age, sex, health status, medical condition, and weight; the nature and severity of symptoms; the type of concurrent treatment; the frequency of treatment; the route of administration; the patient's renal and hepatic function; and the desired effect, as well as the pharmacodynamic characteristics of the specific drug and its mode and route of administration. Clinicians can determine and prescribe the effective dose of the drug necessary to prevent, counteract, or halt the progression of a disease or disorder.

[0126] As a general guideline, the daily oral dose of each active ingredient, when used for the indicated effect, is between about 0.001 and about 1000 mg / kg body weight, preferably between about 0.01 and about 100 mg / kg body weight / day, and most preferably between about 0.1 and about 20 mg / kg / day. In some embodiments, the compound of formula (I) may be administered in doses of about 10 mg / day to about 200 mg / day. In some embodiments, the compound of formula (I) may be administered in doses of approximately 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 110 mg / day, 120 mg / day, 130 mg / day, 140 mg / day, 150 mg / day, 160 mg / day, 170 mg / day, 180 mg / day, 190 mg / day, or 200 mg / day. The dose may be any value or a partial range within the listed ranges.

[0127] Depending on the patient's condition and the intended therapeutic effect, the frequency of administration of the therapeutic agent may vary, for example, from once to six times per day. That is, the frequency of administration may be QD, i.e., once per day; BID, i.e., twice per day; TID, i.e., three times per day; QID, i.e., four times per day; five times / day or six times / day. In another embodiment, the frequency of administration may be BIW, i.e., twice per week; TIW, i.e., three times per week; or QIW, i.e., four times per week.

[0128] Depending on the patient's condition and the intended therapeutic effect, the treatment cycle may include periods during which the drug is not administered. As used herein, “interval administration” refers to the administration of the drug and the subsequent elimination day or week. For example, the treatment cycle may be three weeks long, including two weeks of administration of the drug followed by one week without administration. In some embodiments, the treatment cycle is four weeks long, including three weeks of administration followed by one week without administration of the drug.

[0129] As used herein, the term “treatment cycle” refers to a predetermined period of time for administering a therapeutic agent. Typically, the patient is examined at the end of each treatment cycle to assess the therapeutic effect.

[0130] In one embodiment, each treatment cycle has a duration of approximately 3 days or more. In another embodiment, each treatment cycle is approximately 3 to 60 days. In another embodiment, each treatment cycle is approximately 5 to 50 days. In another embodiment, each treatment cycle is approximately 7 to 28 days. In another embodiment, each treatment cycle is 28 days. In one embodiment, the treatment cycle is approximately 29 days. In another embodiment, the treatment cycle is approximately 30 days. In another embodiment, the treatment cycle is approximately 31 days. In another embodiment, the treatment cycle has a duration of approximately 1 month. In another embodiment, the treatment cycle is of any length from 3 to 8 weeks. In another embodiment, the treatment cycle is of any length from 3 to 6 weeks. In yet another embodiment, the treatment cycle is 3 weeks. In another embodiment, the treatment cycle is 1 month. In another embodiment, the treatment cycle is 4 weeks. In another embodiment, the treatment cycle is 5 weeks. In another embodiment, the treatment cycle is 6 weeks. In another embodiment, the treatment cycle is 7 weeks. In another embodiment, the treatment cycle is 8 weeks. The duration of the treatment cycle may include any value or subrange within the enumerated range, including the endpoint.

[0131] As used herein, the term “co-administration” refers to the coordinated administration of (a) an additional therapeutic agent and (b) a compound of formula (I), or its salts, solvates, esters, and / or prodrugs. For example, co-administration may be simultaneous administration, sequential administration, overlapping administration, interval administration, consecutive administration, or a combination thereof.

[0132] In some embodiments, the administration regimen for the compound of formula (I) is once daily over a continuous 28-day cycle. In some embodiments, the once-daily administration regimen for the compound of formula (I) may be, but are not limited to, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, or 60 mg / day. The compound of formula (I) may be administered once daily in any dose from 20 mg to 60 mg. The dose may be any value or a sub-range within the enumerated range.

[0133] In some embodiments, the administration regimen for the compound of formula (I) is twice daily over a continuous 28-day cycle. In some embodiments, the twice-daily administration regimen for the compound of formula (I) may be, but are not limited to, 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, or 100 mg / day. The compound of formula (I) can be administered twice daily in any dose from 20 mg to 80 mg. In some embodiments, the compound of formula (I) may be administered in any dose from 10 mg / day to 100 mg / day. The dose may be any value or a sub-range within the enumerated range.

[0134] In some embodiments, the administration regimen of the compound of formula (I) may be 20 mg to 60 mg / day once daily for two weeks, followed by a one-week break during the six-week period (e.g., two weeks of administration, one week of no administration). In some embodiments, the administration regimen of the compound of formula (I) may be 10 mg to 100 mg twice daily for two weeks, followed by a one-week break during the six-week period (e.g., two weeks of administration, one week of no administration).

[0135] In some embodiments, the administration regimen of the compound of formula (I) may be once daily at a dose of 20 mg to 60 mg per day for 3 weeks, followed by a 1-week break during the 8-week period (e.g., 3 weeks of administration, 1 week of no administration). In some embodiments, the administration regimen of the compound of formula (I) may be twice daily at a dose of 10 mg to 100 mg per day for 3 weeks, followed by a 1-week break during the 8-week period (e.g., 8 weeks of administration, 1 week of no administration).

[0136] In some embodiments, the administration regimen of the compound of formula (I) may be twice daily on days 1 and 2, weekly for 8 weeks. In some embodiments, the dosage of the compound of formula (I) may be, but is not limited to, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg.

[0137] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily in a three-week cycle, which includes administering the compound for two weeks, followed by a one-week period without administering the compound.

[0138] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily in a four-week cycle, which includes administering the compound for three weeks, followed by a one-week period without administration of the compound.

[0139] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of 6 weeks. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered over a period of 8 weeks.

[0140] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered three times a week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first, third, and fifth days of the week.

[0141] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered four times a week.

[0142] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in a three-week cycle, which includes administering the compound for two weeks, followed by a one-week period without administration of the compound.

[0143] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in a 4-week cycle, which includes administering the compound for 3 weeks, followed by a 1-week period without administration of the compound.

[0144] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily for two days a week. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of eight weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered on the first and second days of each week.

[0145] If the compound of formula I is administered multiple times a week, the dose may be administered on any day or combination of days within the week. For example, a three-times-a-week administration may include administrations on days 1, 3, and 5; days 1, 2, and 3; days 1, 3, and 5; and so on. A two-days-a-week administration may include administrations on days 1 and 2; days 1 and 3; days 1 and 4; days 1 and 5; days 1 and 6; days 1 and 7; and so on.

[0146] cancer In some embodiments, the cancer has a G12C KRAS mutation. In some embodiments, the cancer has a G12D KRAS mutation. In some embodiments, the cancer has a G12RKRAS mutation. In some embodiments, the cancer has a G12S KRAS mutation. In some embodiments, the cancer has a G12V KRAS mutation. In some embodiments, the cancer has a G12WKRAS mutation. In some embodiments, the cancer has a G13D KRAS mutation. In some embodiments, the cancer has an H95D KRAS mutation. In some embodiments, the cancer has an H95Q KRAS mutation. In some embodiments, the cancer has an H95RKRAS mutation. In some embodiments, the cancer has a Q61H KRAS mutation. In some embodiments, the cancer has a G12D KRAS mutation. In some embodiments, the cancer has a Q61K KRAS mutation. In some embodiments, the cancer has a Q61RNRAS mutation. In some embodiments, the cancer has an R68S KRAS mutation.

[0147] In some embodiments, the cancer is non-small cell lung cancer (NSCLC). In some embodiments, the NSCLC is KRAS G12C mutant NSCLC. In some embodiments, the NSCLC is KRAS G12D mutant NSCLC. In some embodiments, the NSCLC is KRAS G12S mutant NSCLC. In some embodiments, the NSCLC is KRAS G12V mutant NSCLC. In some embodiments, the NSCLC is KRAS G13D mutant NSCLC. In some embodiments, the NSCLC is KRAS Q61H mutant NSCLC. In some embodiments, the NSCLC is KRAS Q61K mutant NSCLC. In some embodiments, the NSCLC is KRAS G12R mutant NSCLC. In some embodiments, the NSCLC is KRAS G12W mutant NSCLC. In some embodiments, the NSCLC is KRAS H95D mutant NSCLC. In some embodiments, NSCLC is the KRAS H95Q mutant NSCLC. In some embodiments, NSCLC is the KRAS H95R mutant NSCLC. In some embodiments, NSCLC is the KRAS G12D mutant NSCLC. In some embodiments, NSCLC is the KRAS R68S mutant NSCLC.

[0148] In some embodiments, the cancer is aKRAS-treated G12C NSCLC. In some embodiments, the cancer is KRAS-treated G12D NSCLC. In some embodiments, the cancer is KRAS-treated G12SNSCLC. In some embodiments, the cancer is aKRAS-treated G12V NSCLC. In some embodiments, the cancer is aKRAS-treated G13D NSCLC. In some embodiments, the cancer is KRAS-treated Q61HNSCLC. In some embodiments, the cancer is KRAS-treated Q61KNSCLC. In some embodiments, the cancer is NRAS-treated Q61R NSCLC. In some embodiments, the cancer is aKRAS-treated G12R NSCLC. In some embodiments, the cancer is KRAS-treated G12WNSCLC. In some embodiments, the cancer is KRAS-treated H95D NSCLC. In some embodiments, the cancer is KRAS-treated H95Q NSCLC. In some embodiments, the cancer is aKRAS-treated H95R NSCLC. In some embodiments, the cancer is KRAS-treated G12D NSCLC. In some embodiments, the cancer is KRAS-treated R68S NSCLC.

[0149] In some embodiments, the cancer is colorectal cancer (CRC). In some embodiments, the CRC is a KRAS variant CRC. In some embodiments, the CRC is a KRAS G12C variant CRC. In some embodiments, the CRC is a KRAS G12D variant CRC. In some embodiments, the CRC is a KRAS G12S variant CRC. In some embodiments, the CRC is a KRAS G12V variant CRC. In some embodiments, the CRC is a KRAS G13D variant CRC. In some embodiments, the CRC is a KRAS Q61H variant CRC. In some embodiments, the CRC is a KRAS Q61K variant CRC. In some embodiments, the CRC is an aNRAS variant CRC. In some embodiments, the CRC is an NRAS Q61R variant CRC.

[0150] In some embodiments, the cancer has one or more acquired mutations. In some embodiments, the acquired mutations result from a first-line treatment. In some embodiments, the first-line treatment is a KRAS inhibitor. In some embodiments, the KRAS inhibitor is a KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is adagrasib. In some embodiments, the KRAS G12C inhibitor is sotorasib. In some embodiments, the cancer is a solid tumor carcinoma. In some embodiments, the cancer is NSCLC.

[0151] In some embodiments, the acquired mutation is an acquired KRAS mutation. In some embodiments, the acquired mutation is KRAS G12C. In some embodiments, the acquired mutation is KRAS G12D. In some embodiments, the acquired mutation is KRAS G12R. In some embodiments, the acquired mutation is KRAS G12V. In some embodiments, the acquired mutation is KRAS G12W. In some embodiments, the acquired mutation is KRAS G13D. In some embodiments, the acquired mutation is KRAS H95D. In some embodiments, the acquired mutation is KRAS H95D. In some embodiments, the acquired mutation is KRAS H95Q. In some embodiments, the acquired mutation is KRAS H95R. In some embodiments, the acquired mutation is KRAS Q61H. In some embodiments, the acquired mutation is KRAS R68S.

[0152] In some embodiments, the acquired mutation is an acquired MAPK pathway mutation. In some embodiments, the acquired MAPK pathway mutation is MAP2K1 K57N. In some embodiments, the acquired MAPK pathway mutation is MAP2K1 K57T. In some embodiments, the acquired MAPK pathway mutation is CCDC6-RET. In some embodiments, the acquired MAPK pathway mutation is RITI P128L. In some embodiments, the acquired MAPK pathway mutation is PTEN G209V. In some embodiments, the acquired MAPK pathway mutation is BRAF V600E. In some embodiments, the acquired MAPK pathway mutation is MAP2K1 199_K104del. In some embodiments, the acquired MAPK pathway mutation is MAP2K1 K57N. In some embodiments, the acquired MAPK pathway mutation is EML4-ALK. In some embodiments, the acquired MAPK pathway mutation is EGFR A289A. In some embodiments, the acquired MAPK pathway mutation is FGFR3-TACC3. In some embodiments, the acquired MAPK pathway mutation is AKAP9-BRAF. In some embodiments, the acquired MAPK pathway mutation is RAFI-CCDC176. In some embodiments, the acquired MAPK pathway mutation is RAF1-TRAK1. In some embodiments, the acquired MAPK pathway mutation is NRAS Q61K. In some embodiments, the acquired MAPK pathway mutation is MAP2K1 E102 1103DEL. In some embodiments, the acquired MAPK pathway mutation is NRF1-BRAF.

[0153] In some embodiments, the acquired mutation is a KRAS G12C reactivation mutation. In some embodiments, the KRAS G12C reactivation mutation is an RKRAS G12C gene amplification. In some embodiments, the KRAS G12C reactivation mutation is NF1 R22637(LoF).

[0154] In some embodiments, the acquired mutation is a non-G12C activated KRAS mutation. In some embodiments, the non-G12C activated KRAS mutation is KRAS G12D. In some embodiments, the non-G12C activated KRAS mutation is KRAS G12R. In some embodiments, the non-G12C activated KRAS mutation is KRAS G12V. In some embodiments, the non-G12C activated KRAS mutation is KRAS G12W. In some embodiments, the non-G12C activated KRAS mutation is KRAS G13D. In some embodiments, the non-G12C activated KRAS mutation is KRAS Q61H. In some embodiments, the non-G12C activated KRAS mutation is KRAS Q61K.

[0155] In some embodiments, the acquired mutation is a hindering KRAS G12C mutation. In some embodiments, the hindering KRAS G12C mutation is KRAS R68S. In some embodiments, the hindering KRAS G12C mutation is KRAS H95D. In some embodiments, the hindering KRAS G12C mutation is KRAS H95Q. In some embodiments, the hindering KRAS G12C mutation is KRAS H95R. In some embodiments, the hindering KRAS G12C mutation is KRAS Y96C.

[0156] In some embodiments, the acquired mutation is an RTK-activating mutation. In some embodiments, the RTK-activating mutation is EGFR A289V. In some embodiments, the RTK-activating mutation is RET M918T. In some embodiments, the RTK-activating mutation is MET gene amplification. In some embodiments, the RTK-activating mutation is EML-ALK. In some embodiments, the RTK-activating mutation is CCDC6-RET. In some embodiments, the RTK-activating mutation is FGFR3-TACC3.

[0157] In some embodiments, the acquired mutation is a downstream RAS / MAPK activating mutation. In some embodiments, the downstream RAS / MAPK activating mutation is BRAF V600E. In some embodiments, the downstream RAS / MAPK activating mutation is MAP2K I99_K104del. In some embodiments, the downstream RAS / MAPK activating mutation is MAP2K1 I99_K104del. In some embodiments, the downstream RAS / MAPK activating mutation is MAP2K1 E102_I103del. In some embodiments, the downstream RAS / MAPK activating mutation is an RAF fusion.

[0158] In some embodiments, the acquired mutation is a parallel pathway activation mutation. In some embodiments, the parallel pathway activation mutation is PIK3CA H1047R. In some embodiments, the parallel pathway activation mutation is PIK3R1S361fs. In some embodiments, the parallel pathway activation mutation is PTENN48K. In some embodiments, the parallel pathway activation mutation is PTEN G209V. In some embodiments, the parallel pathway activation mutation is RIT1P128L.

[0159] Kits and products Some embodiments relate to kits and products comprising a compound of formula I and / or at least one KRAS G12C inhibitor. For example, a kit or product may include a package or container having the compound of formula I. Such kits and products may further include a product insert or label having approved drug administration and indication information, including a method of using the compound of formula I in combination with a separately provided KRAS G12C inhibitor. The kit can be used in a method of treating cancer as described herein.

[0160] In some embodiments, a kit or product may include both a compound of formula I and at least one KRAS G12C inhibitor. In some embodiments, the KRAS G12C inhibitor is, for example, AMG510. In some embodiments, the KRAS G12C inhibitor is, for example, MRTX849. Such a kit may include one or more containers or packages, one or more containers or packages may include one or both combination drugs together in a single container and / or package, or in separate packages / containers. In some cases, the two drugs are packaged separately but contained in a single package, container or box. Such kits and products may further include a product insert or label having approved drug administration and indication information, including a method of using the compound of formula I in combination with a KRAS G12C inhibitor. The kit may be used in a method of treating cancer as described herein. [Examples]

[0161] General procedure All starting materials and solvents were obtained from commercial sources or prepared according to literature references.

[0162] Example 1 This embodiment demonstrates a synergistic combination of the compound of formula I and a KRAS G12C inhibitor.

[0163] Cell proliferation assay Cells (2000 cells per well) were seeded in 100 μl of cell culture medium on a 96-well plate and treated with compound I alone or with a fixed concentration of AMG510. On day 5, 50 μl of CellTiter-Glo (CTG) reagent (Promega) was added, and the plates were cultured for 10 minutes with gentle shaking. After 10 minutes of culture, the luminescence signal was determined according to the manufacturer's instructions (Promega), and the graph was plotted using Prism GraphPad.

[0164] Combined cell proliferation assay Cells (2000 cells per well) were seeded in 100 μl of cell culture medium on a 96-well plate. Cells were treated with AMG510 and the compound of formula I at concentrations ranging from 0 to 10 μM using the Tecan D300e Digital Dispenser combination matrix protocol. On day 5, 50 μl of CellTiter-Glo (CTG) reagent (Promega) was added, and the plate was cultured for 10 minutes with gentle shaking. After 10 minutes of culture, the luminescence signal was determined according to the manufacturer's instructions (Promega), and combination data was generated using the standard HSA model with Combinenefit software. Combination synergies are represented by positive numbers in the results table. Negative numbers represent combination antagonisms.

[0165] The results of these experiments are shown in Figures 1-4, and will be discussed further below.

[0166] Figure 1 shows data demonstrating that the compound of formula I and AMG510 synergistically bind to inhibit cell proliferation in NCI-H358 cells with the KRAS G12C mutation.

[0167] Figure 2A shows plots of activity percentage versus inhibitor concentration (logM) in NCI-14358 cells treated with the compound of formula I alone and in combination with AMG510. IC50 data for NCI-H358 cells treated with the compound of formula I alone and in combination with AMG510 are shown in the table.

[0168] Figure 2B shows a bar graph of CTG activity percentages, indicating that AMG510(InM) alone did not reduce cell viability in NCI-H358 cells.

[0169] Figure 3 shows data demonstrating that the compound of formula I and AMG510 synergistically bind to inhibit cell proliferation in the KRAS G12C mutant NCI-H2122.

[0170] Figure 4A shows plots of activity percentage versus inhibitor concentration (logM) in NCI-142122 cells treated with the compound of formula I alone and in combination with various concentrations of AMG510.

[0171] Figure 4B shows a bar graph of CTG activity percentages, indicating that AMG510(InM) alone did not reduce cell viability in NCI-H2122 cells.

[0172] In summary, this dataset demonstrates that the combination of the compound of formula I with a KRAS G12C inhibitor provides synergistic inhibition of KRAS G12C-mutated cancer cell viability. The activity of the compound of formula I can be synergistically enhanced by combining it with a KRAS G12C inhibitor in cells with KRAS G12C mutations.

[0173] Example 2 The results of these experiments are shown in Figures 7-10, and will be discussed further below.

[0174] Figure 5 shows a matrix representation of HSA synergies and antagonists, demonstrating that the compound of formula I and adagrasib are synergistically combined to inhibit cell proliferation in KRAS G12C mutant NCI-H358 cells.

[0175] Figure 6A shows a plot of activity percentage versus inhibitor concentration (logM) in NCI-1358 cells treated with the compound of formula I alone (black circles) and in combination with 1 nM adagrasib (black squares).

[0176] Figure 6B shows a bar graph of CTG activity percentages, indicating that 1 nM adagrasib alone did not reduce cell viability in NCI-H358 cells.

[0177] [Table 1]

[0178] Figure 7 shows a matrix representation of HSA synergistic and antagonistic effects, demonstrating that the compound of formula I and adagrasib are synergistically combined to inhibit cell proliferation in KRAS G12C mutant NCI-H2122 cells.

[0179] Figure 8 shows a matrix representation of HSA synergistic and antagonistic effects, demonstrating that the compound of formula I and adagrasib are synergistically combined to inhibit cell proliferation in KRAS G12C mutant KYSE-410 cells.

[0180] Figure 9A shows plots of activity percentage versus inhibitor concentration (logM) in NCI-H2122 cells treated with adagrasib compound I alone (black circles, line 1) and in combination with 1 nM (black squares, line 2), 5 nM (black circles, line 3), or 10 nM (black squares, line 4).

[0181] Figure 9B shows bar graphs of percentage CTG activity, indicating that adagrasib alone at 1 nM, 5 nM, or 10 nM concentrations did not reduce cell viability in NCI-H2122 cells.

[0182] [Table 2]

[0183] Figure 10A shows the tumor volume (mm²) of a KRAS G12C mutant CRC022 PDX tumor xenograft model treated with the vehicle (black circle, line 1), adagrasib alone (30 mg / kg QD, solid triangle, line 2), the compound of formula I alone at 10 mg / kg / dose BID (black circle, line 3), the compound of formula I at 30 mg / kg QD (black triangle, line 4), a combination of the compound of formula I (10 mg / kg / dose BID) and adagrasib at 30 mg / kg QD (black circle, line 5), and a combination of the compound of formula I (30 mg / kg QD) and adagrasib at 30 mg / kg QD (black triangle, line 6). 3 This shows a plot of the relationship between the treatment period (days) and the treatment period.

[0184] Figure 10B shows the tumor volume (mm²) of KRAS G12C mutant H2122 CDX tumor xenograft models treated with the vehicle (black circle, line 1), adagrasib alone (30 mg / kg QD, solid triangle, line 2), compound of formula I alone (10 mg / kg / dose BID, black circle, line 3), compound of formula I alone at 30 mg / kg QD (black triangle, line 4), compound of formula I (10 mg / kg / dose BID) and adagrasib at 30 mg / kg QD (black circle, line 5), and compound of formula I (30 mg / kg QD) and adagrasib at 30 mg / kg QD (black triangle, line 6). 3 This shows a plot of the relationship between the treatment period (days) and the treatment period.

[0185] Example 3: The combination of the compound of formula I and adagrasib is KRAS G12C It showed a synergistic effect on cell proliferation in cells. Cell lines were obtained from ATCC (NCI-H358#CRL-5807 and NCI-H2122#CRL-5985). KYSE-410 was obtained from Millipore Sigma (#94072023). Cells were cultured in RPMI containing 10% FBS and penicillion / stremtomycin and maintained at 37°C / 5% CO2.

[0186] Cell proliferation assay Cells (2000 cells per well) were seeded in 100 μl of cell culture medium in a 96-well plate. Using the Tecan D300e Digital Dispenser combination matrix protocol, cells were treated with the compound of formula I and adagrasib at concentrations ranging from 0 to 10 μM. On day 5, 50 μl of CellTiter-Glo (CTG) reagent (Promega) was added, and the plates were cultured for 10 minutes with gentle shaking. After 10 minutes of culture, the luminescence signal was determined according to the manufacturer's instructions (Promega), and combined data was generated using Combinefit software.

[0187] Adagrasib-sensitive cells, NCI-H358, were dispensed into 96-well plates. After overnight incubation, the compound of formula I and adagrasib were added to the cells using the Tecan D300e Digital Dispenser combination matrix protocol. After 5 days of incubation, the CellTiter-Glo assay was performed, and the combined synergistic effect was calculated using the standard HSA model with Combinefit software. Data were represented by the area and intensity of color codes, with synergistic effects represented by blue, additives by green, and antagonistic effects by red. The combined data were analyzed using NSCLC KRAS. G12C A synergistic effect on cell viability was observed in mutant cells NCI-H358 (Figure 5).

[0188] The combinations were studied by cell viability assays. NCI-H358 cells were dispensed into 96-well plates. After overnight culture, cells were treated with either the compound of formula I alone or in combination with the compound of formula I and fixed final concentration adagrasib (1 nM), and the CellTiter-Glo assay was performed after 5 days. Treatment with adagrasib (1 nM) alone did not inhibit cell viability in NCI-H358 (Figure 6A). However, fixed concentration adagrasib (1 nM) increased the sensitivity of NCI-H358 cells to the compound of formula I (Figure 6A). The IC50 of the compound of formula I decreased to approximately one-third after treatment with a final concentration of 1 nM adagrasib.

[0189] KRAS G12C KRAS is moderately sensitive to inhibitors. G12CTo maximize the therapeutic response in cells, the inventors tested combinations of compound I and adagrasib in NCI-H2122 and KYSE-410 cells. Cells were dispensed into 96-well plates. After overnight incubation, compound I and adagrasib were added to the cells using the Tecan D300e Digital Dispenser combination matrix protocol. After 5 days of incubation, the CellTiter-Glo assay was performed, and the combined synergy was calculated using the standard HSA model with Combinefit software. Data are represented by the area and intensity of color codes, with synergy represented by dark gray, additive by light gray, and antagonist by gray. Combination data were obtained for NCI-H2122 and KRAS G12C The drug showed a synergistic effect on cell viability in mutated KYSE-410 cells (Figures 7 and 8).

[0190] The combinations were confirmed by a follow-up viability assay. NCI-H2122 cells were dispensed onto 96-well plates. After overnight culture, cells were treated with either the compound of formula I alone or a combination of the compound of formula I and adagrasib at final fixation concentrations (1, 5, and 10 nM), and the CellTiter-Glo assay was performed after 5 days. Treatment with adagrasib alone showed less than 10% inhibition of cell viability in NCI-H2122 (Figure 9B), while fixation concentrations of adagrasib dose-dependently increased the sensitivity of the compound of formula I in NCI-H2122 cells (Figure 9A). The IC50 of the compound of formula I decreased dose-dependently from 2785 nM to 190 nM with adagrasib treatment. Overall, the data suggest that the combination of the compound of formula I and adagrasib is beneficial for KRAS G12C This suggests that it is effective in mutant human cancer cells.

[0191] Example 4 - Phase 1b / 2 trial of a drug targeting the mitogenic factor-activated protein kinase pathway in a patient with advanced non-small cell lung cancer. This study includes: 1) evaluating the safety and tolerability of the formula I compound in combination with other cancer therapies in study participants with advanced non-small cell lung cancer (NSCLC); 2) determining the maximum tolerated dose (MTD) and / or recommended dose (RD) of the formula I compound when administered in combination with other cancer therapies; 3) evaluating the antitumor activity of the formula I compound in combination with other cancer therapies; and 4) evaluating the pharmacokinetic (PK) profiles of the formula I compound and other cancer therapies when administered in combination.

[0192] The Phase 1b / 2 trial will include evaluating the safety, tolerability, and antitumor activity of compound I in combination with other cancer therapies in trial participants with advanced NSCLC. The study will include a dose-escalation cohort in which compound I and sotorasib will be administered to trial participants with advanced NSCLC bearing the Kirsten rat sarcoma G12C mutation (KRAS G12Cm). Compound I will be administered orally in combination with sotorasib to study participants with KRAS G12Cm NSCLC at successively escalating doses until unacceptable toxicity, disease progression, or withdrawal of consent. As dose escalation continues, the compound I administered orally at the RD identified from each dose-escalation cohort will be evaluated in trial participants with advanced EGFRm or KRAS G12Cm NSCLC.

[0193] Inclusion criteria Age: 18 years or older.

[0194] They are willing and capable of providing written informed consent.

[0195] Histologically or cytologically confirmed NSCLC is associated with the presence of EGFR mutations that make the patient sensitive to EGFR inhibitors, or KRAS G12C mutations.

[0196] Response Criteria for Solid Tumors (RECIST) v1.1.

[0197] Appropriate bone marrow and organ function. Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. Grade 0: Fully active and capable of performing all pre-disease outcomes without limitation; Grade 1: Physically strenuous activities are limited, but the person is able to walk and perform simple, sedentary tasks such as light housework or office work.

[0198] We are willing to follow all visits, assessments, and procedures required by all protocols.

[0199] They can swallow oral medication.

[0200] Exclusion criteria Concurrent treatment with any systemic anticancer therapy for NSCLC, including any approved drug or investigational drug.

[0201] For participants with KRAS G12Cm NSCLC, prior treatment with SHP2, ERK, or a KRAS G12C inhibitor (depending on which cohort is being considered for enrollment).

[0202] Palliative radiotherapy within 7 days of registration.

[0203] History of unacceptable toxicity to treatment with sotorasib.

[0204] Major surgery performed within 28 days of registration.

[0205] Unresolved toxicity from prior systemic therapy exceeding NCI Common Terminology Criteria for Adverse Events (CTCAE) Grade 1 at the time of enrollment, excluding toxicity not considered a safety risk (e.g., alopecia, vitiligo, and Grade 2 neuropathy due to prior chemotherapy).

[0206] A history of another malignant tumor within five years of the first administration, except for patients who have been disease-free for more than two years after treatment with curative intent, or who have a carcinoma in situ.

[0207] Except for patients who have completed definitive therapy (surgery or radiotherapy), symptomatic and unstable brain metastases or spinal cord compressions are defined as those who have not received steroids and have a stable neurological condition for at least two weeks after completion of definitive therapy and steroids.

[0208] Interstitial lung disease (ILD), drug-induced ILD, or a history of radiation pneumonitis or clinically active ILD requiring steroid treatment.

[0209] Cardiovascular dysfunction or clinically significant cardiovascular disease.

[0210] Retinal pigment epithelial detachment (RPED), central serous retinopathy, retinal vein occlusion (RVO), or a history or current evidence of predisposition to RPED or RVO.

[0211] Evidence of a serious or uncontrolled systemic disease, or any other significant clinical impairment or laboratory findings that would make the patient unsuitable to participate in the study.

[0212] Pregnant or breastfeeding women.

[0213] Contraindications to the use of sotorasib on a local label basis.

[0214] Example 5 - In vivo studies of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model SW1573. material and method Vehicle / control samples were prepared in 100 mM acetic acid in deionized water, adjusted to pH 4.8-5.0, and stored under ambient conditions throughout a 27-day administration period in mice. Test samples of the compound of Formula 1 were prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination agent sotarasib was prepared in a vehicle of 50% / 50% w / w PEG400 / PG, adjusted weekly with HCl, and stored at 2-8°C.

[0215] Female Balb / c nude mice were purchased from Vital River (Beijing, China). The mice were 6–8 weeks old at the time of transplantation. The mice were moved to an animal room in an animal facility and allowed to acclimate to their new environment for at least 3 days before the start of any experiment. All procedures related to animal handling, care, and treatment in this study were carried out in accordance with guidelines approved by the Institutional Animal Care and Use Committee (IACUC) of GenenDesign (Shanghai, China). Furthermore, all parts of this study were conducted in accordance with research protocols approved by the research director and applicable standard operating procedures (SOPs) at GenenDesign.

[0216] CDX preparation SW1573 was a human NSCLC cell line carrying the KRAS G12C mutation. The cell line was purchased from ATCC. Early passaged SW1573 cells were maintained in vitro as a monolayer culture at 37°C in L-15 medium supplemented with 10% fetal bovine serum (FBS) in 5% CO2 in the air. The medium was changed every 2-3 days, and tumor cells were routinely passaged with trypsin-EDTA at 80-90% confluence, not exceeding 5 passages. Cells growing in the exponential growth phase were harvested and counted for tumor inoculation into mice.

[0217] Before subcutaneous transplantation, mice were anesthetized with isoflurane. 5 × 10⁶ grafts were mixed with 50% Matrigel. 6A 200 μL cell suspension containing SW1573 tumor cells was subcutaneously transplanted into the right flank of mice using a syringe. Animal health and tumor growth were monitored daily after transplantation. Tumor volume was measured twice weekly using a caliper if the xenografted tumor was palpable and measurable. The average subcutaneous tumor volume was approximately 200 mm³. 3 (150~263mm 3 When the tumor size reached a certain range, mice with tumors were randomized to different groups (n=8 mice / group). The randomization day is denoted as treatment day 0.

[0218] treatment Mice with tumors were treated on the day of randomization. The treatment start day was designated as treatment day 0. Mice were administered orally as a vehicle control, monotherapy with the compound of formula I at 10 mg / kg / dose BID and 30 mg / kg QD, and monotherapy with sotorasib at 30 and 100 mg / kg QD. Mice were also treated in two combination treatment groups of the compound of formula I and sotorasib: one group received the combination of the compound of formula I at 10 mg / kg / dose BID and sotorasib at 100 mg / kg QD, and the other group received the combination of the compound of formula I at 30 mg / kg QD and sotorasib at 100 mg / kg QD. The dose volume was 5 mL / kg, and the interval between BID regimens was 8 hours. This study was terminated when the termination criteria defined in the study protocol were met.

[0219] result Figure 11 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model SW1573. No significant changes in body weight were observed in either the control or treatment groups.

[0220] knot As shown in Figure 11, the combination of the compound of formula I and sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant NSCLC CDX model SW1573.

[0221] Example 6: In vivo studies of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model, NCI-H358. material and method Vehicle / control: 100 mM acetic acid in deionized water was prepared by adjusting the pH to 4.8-5.0 and stored under ambient conditions throughout 28 days of administration to mice. Test samples of the compound of formula I were prepared in a vehicle of 100 mM acetic acid buffer throughout 28 days of administration and stored under ambient conditions. The combination agent sotarasib was prepared in a vehicle of 50% PEG400 / 50% PG, adjusted with HCl throughout 28 days of administration and stored at 2-8°C.

[0222] Female nude (Nu / Nu) mice were purchased from Jackson Laboratory (USA). The mice were 6-8 weeks old at the time of transplantation. The mice were moved to an animal room in the animal facility and allowed to acclimate to the new environment for 3 days before the start of any experiment.

[0223] CDX preparation NCI-H358 was a human NSCLC cell line carrying the KRAS G12C mutation. The cell line was purchased from ATCC. Early passaged NCI-H358 cells were maintained in vitro as monolayer cultures in RPMI1640 medium supplemented with 10% fetal bovine serum (FBS) at 37°C in 5% CO2 air. The medium was refreshed every 2-3 days, and tumor cells were routinely passaged at 80-90% confluence with trypsin-EDTA, not exceeding 5 passages. Cells growing in the exponential growth phase were harvested and counted for tumor inoculation into mice.

[0224] Before subcutaneous transplantation, mice were anesthetized with isoflurane. 5 × 10⁶ grafts were mixed with 50% Matrigel.6 A 200 μL cell suspension containing NCI-H358 tumor cells was subcutaneously transplanted into the right flank of mice using a syringe. The animals' health and tumor growth were monitored daily after transplantation. Tumor volume was measured twice weekly using a caliper if the xenografted tumor was palpable and measurable. Subcutaneous tumor volume was approximately 300 mm³. 3 (150~390mm 3 When the mean of the range was reached, mice with tumors were randomized to different groups (n=10 mice / group). The randomization day is denoted as treatment day 0.

[0225] treatment Treatment was initiated the day after randomization. The treatment initiation day was designated as Treatment Day 1. Mice were administered orally by vehicle, monotherapy with the compound of formula I at 10 mg / kg / dose BID and 30 mg / kg QD, and monotherapy with sotorasib at 10 mg / kg QD. Mice were also treated in two combination treatment groups: one group received the compound of formula I at 10 mg / kg / dose BID and sotorasib at 10 mg / kg QD, and the other group received the compound of formula I at 30 mg / kg QD and sotorasib at 10 mg / kg QD. The administration volume was 5 mL / kg, and the interval between BID regimens was 8 hours. The compound of formula I was administered first, followed by sotorasib 1 hour later in the combination treatment groups. This study was terminated when the termination criteria defined in the study protocol were met.

[0226] result Figure 12 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model NCI-H358. No significant changes in body weight were observed in either the control or treatment groups.

[0227] knot As shown in Figure 12, the combination of the compound of formula I and sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant NSCLC CDX model NCI-H358.

[0228] Example 7: In vivo studies of formula I compound alone, sotorasib alone, and formula 1 in combination with sotorasib in the KRAS G12C mutant esophageal squamous cell carcinoma CDX model, KYSE-410. material and method Vehicle / control: 100 mM acetic acid in deionized water was prepared by adjusting the pH to 4.8-5.0 and stored under ambient conditions throughout 21 days of administration in mice. Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination agent sotorasib was prepared in a vehicle of 50% w / w polyethylene glycol 400 (PEG400) and 50% w / w propylene glycol (PG) and stored at 2-8°C.

[0229] Female nude (Nu / Nu) mice were purchased from Jackson Laboratory (USA). The mice were 6-7 weeks old at the time of transplantation. The mice were moved to the animal room of the animal facility and allowed to acclimate to their new environment for at least 3 days before the start of any experiment. All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with guidelines approved by the Institutional Animal Care and Use Committee (IACUC) of Explora BioLabs (San Diego, California). In addition, all parts of this study conducted at Explora BioLabs adhered to research protocols approved by the research director and applicable standard operating procedures (SOPs).

[0230] CDX preparation KYSE-410, a human esophageal squamous cell carcinoma cell line carrying the KRAS G12C mutation, was purchased from ATCC and cultured in a medium containing RPMI-1640 10% fetal bovine serum (FBS) at 37°C in 5% CO2 in air. The medium was changed every 2-3 days, and tumor cells were routinely subcultured with trypsin-EDTA at 80-90% confluence. Cells growing in the exponential growth phase were harvested and counted for inoculation.

[0231] Prior to subcutaneous transplantation, mice were anesthetized with isoflurane. A 200 μL cell suspension containing 4x10⁶ KYSE-410 tumor cells mixed with 50% Matrigel was subcutaneously transplanted into the right flank of the mice using a syringe. The health of the animals and tumor growth were monitored daily after transplantation. Tumor volume was measured twice weekly using a caliper if the xenografted tumor was palpable and measurable. The average subcutaneous tumor volume was approximately 196 mm³. 3 (150~300mm 3 When the tumor size reached a certain range, mice with tumors were randomized to different groups (n=10 mice / group). The randomization day is denoted as treatment day 0.

[0232] treatment Treatment was initiated on the day after randomization. The treatment initiation day was designated as treatment day 1. Mice were administered orally by vehicle control solution, compound I of formula I alone at 10 mg / kg / dose BID, compound I of formula I at 30 mg / kg QD, or sotarasib at 100 mg / kg QD. Two additional groups received combination treatment with compound I of formula I and sotarasib; one group received compound I of formula I at 10 mg / kg / dose BID, and the other group received compound I of formula I at 30 mg / kg QD; both combination groups received sotarasib at 100 mg / kg QD. The administration volume was 5 mL / kg, and the interval between BID regimens was 8 hours. In the combination groups, sotarasib was administered 1 hour after the first BID or QD dose of compound I. The study was concluded on treatment day 21 as defined in the study protocol.

[0233] result Figure 13 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant esophageal squamous cell carcinoma CDX model KYSE-410. No significant weight changes were observed in the control or treatment groups.

[0234] knot As shown in Figure 13, the combination of the compound of formula I and sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant esophageal squamous cell carcinoma CDX model KYSE-410.

[0235] Example 8: In vivo studies of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model, CO-04-0310. material and method Vehicle / control: 100 mM acetic acid in deionized water was prepared by adjusting the pH to 4.8-5.0 and stored under ambient conditions throughout 28 days of administration in mice. Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination agent sotorasib was prepared in a vehicle of 50% w / w polyethylene glycol 400 (PEG400) and 50% w / w propylene glycol (PG) and stored at 2-8°C.

[0236] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were transferred to a special pathogen-free (SPF) environment within the barium facility and allowed to acclimate for at least three days before the start of any experiment. All procedures related to animal handling, care, and treatment in this study were carried out in accordance with guidelines approved by the WuXi AppTec Institutional Animal Care and Use Committee (IACUC). During the study, animal care and use were conducted in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). In addition, all parts of this study conducted at WuXi AppTec adhered to research protocols approved by the research director and applicable standard operating procedures (SOPs).

[0237] Preparation of PDX The CO-04-0310 PDX model was established for preclinical efficacy studies at WuXi AppTec. This PDX model was derived from an 82-year-old female Chinese CRC patient. The KRAS G12C mutation in the PDX model CO-04-0310 was confirmed by whole exome sequencing and PCR sequencing. Mouse skin was cleaned with appropriate surgical scrub and alcohol on the right flank. Tumor fragments (15-30 mm) were collected from the PDX model. 3 The tumor was subcutaneously transplanted into the right flank of female Balb / c nude mice using an 18g trocar needle. The tumor size ranged from 100 to 218 mm in volume. 3 When the target was reached, mice with tumors were randomly divided into test groups, with 8 mice in each group. The randomization day is denoted as treatment day 0.

[0238] treatment Treatment was initiated on the day after randomization. The treatment initiation day was designated as treatment day 1. Mice were administered orally with vehicle control solution, compound I of formula I alone at 10 mg / kg / dose BID, compound I of formula I alone at 30 mg / kg QD, and sotarasib at 30 mg / kg QD. Two further groups received combination treatment with compound I of formula I and sotarasib: one group received compound I of formula I at 10 mg / kg / dose BID and sotarasib at 30 mg / kg QD, and the other group received compound I of formula I at 30 mg / kg QD and sotarasib at 30 mg / kg QD. The administration volume was 5 mL / kg, and the interval between BID regimens was 8 hours. This study was concluded on treatment day 28 as defined in the study protocol.

[0239] result Figure 14 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model CO-04-0310. No significant changes in body weight were observed in the control or treatment groups.

[0240] knot As shown in Figure 14, the combination of the compound of formula I and sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant CRC PDX model CO-04-0310.

[0241] Example 9: In vivo studies of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model, CR2528. material and method Vehicle / control: 100 mM acetic acid in deionized water was prepared by adjusting the pH to 4.8-5.0 and stored under ambient conditions throughout 24 days of administration to mice. Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination agent sotorasib was prepared in a vehicle of 50% w / w polyethylene glycol 400 (PEG400) and 50% w / w propylene glycol (PG) and stored at 2-8°C.

[0242] Female Balb / c nude mice were purchased from SPF (Beijing) Laboratory Animal Technology Co. Ltd. (Beijing, China). The mice were 7-9 weeks old at the time of transplantation. The mice were transferred to a specific pathogen-free (SPF) environment in the animal facility and allowed to acclimate to the new environment for at least 3 days before the start of any experiment. All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with guidelines approved by the Institutional Animal Care and Use Committee (IACUC) of Crown Bioscience (Beijing, China). During the study, animal care and use were carried out in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Furthermore, all parts of this study conducted at Crown Bioscience (Beijing, China) adhered to research protocols approved by the research director and applicable standard operating procedures (SOPs).

[0243] Preparation of PDX The CR2528 PDX model was established for preclinical efficacy studies at CrownBio. This PDX model was derived from a 73-year-old male Chinese CRC patient. The KRAS G12C mutation in the PDX model CR2528 was confirmed by both RNA sequencing and exome sequencing. Mouse skin was washed with appropriate surgical scrub and iodine foam on the right abdomen. Tumor fragments (2-3 mm in diameter) collected from the PDX model were subcutaneously transplanted into the right abdomen of female Balb / c nude mice using an 18-g trocar needle. When the average tumor size reached 204 mm 3 (range 149 - 275 mm 3 ), the mice with tumors were randomly divided into six test groups, each group containing 8 mice. The randomization day was designated as treatment day 0.

[0244] Treatment Treatment was initiated on the day of randomization. The treatment start day was set as treatment day 0. The mice were administered by oral gavage with vehicle control solution, sotorasib at 30 mg / kg QD, compound of formula I alone at 10 mg / kg / dose BID, and compound of formula I alone at 30 mg / kg QD. Two additional groups received combination treatment of the compound of formula I and sotorasib, one group was administered the compound of formula I at 10 mg / kg / dose BID and sotorasib at 30 mg / kg QD, and the other group was administered the compound of formula I at 30 mg / kg QD and sotorasib at 30 mg / kg QD. The dosing volume was 5 mL / kg, and the interval for the BID regimen was 8 hours. Sotorasib was administered 1 hour after the QD dosing of the compound of formula I or the first BID dosing in the combination groups.

[0245] Results Figure 15 shows a graph of tumor volume over the treatment period with regimens of the compound of formula I alone, sotorasib alone, and combination of the compound of formula I and sotorasib in the KRAS G12C mutant CRC PDX model CR2528. No significant body weight changes were observed in the control and treatment groups.

[0246] Conclusion As shown in Figure 15, the combination of the compound of formula I and sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant CRC PDX model CR2528.

[0247] Example 10: In vivo studies of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant NSCLC CDX model NCI-H2122. material and method Vehicle / control: 100 mM acetic acid in deionized water was prepared by adjusting the pH to 4.8-5.0 and stored under ambient conditions throughout 14 days of administration to mice. Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination agent sotorasib was prepared in a vehicle of 50% w / w polyethylene glycol 400 (PEG400) and 50% w / w propylene glycol (PG) and stored at 2-8°C.

[0248] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were 6–8 weeks old at the time of transplantation. All procedures related to animal handling, care, and treatment in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Institutional Animal Care and Use Committee (IACUC). The animal facilities and programs operated under the standards of the Guideforthe Care and Use of Laboratory Animals (NRC, 2011) and were accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0249] CDX preparation KRAS G12C The mutant human NSCLC cell line NCI-H2122 was purchased from ATCC and cultured at 37°C in RPML1640 medium containing 10% fetal bovine serum (FBS) with 5% CO2 in the air. The medium was changed every 2-3 days, and tumor cells were routinely subcultured with trypsin-EDTA at 80-90% confluence. Cells growing in the exponential growth phase were harvested and counted for inoculation. Mice were anesthetized with isoflurane before subcutaneous transplantation. 2 × 10⁶ cells were mixed with 50% Matrigel. 6 A 200 μL cell suspension containing NCI-H2122 tumor cells was subcutaneously transplanted into the right flank of mice using a syringe. Animal health and tumor growth were monitored daily after transplantation. Tumor volume was measured twice weekly using a caliper if the xenografted tumor was palpable and measurable. The average subcutaneous tumor volume was approximately 190 mm³. 3 (146~262mm 3 When the tumor reached a certain range, mice with tumors were randomized to the test group (n=8 mice / group). The randomization day is denoted as treatment day 0.

[0250] treatment Treatment was initiated on the day of randomization. The treatment start day was designated as treatment day 0. Mice were administered either orally with a vehicle control solution or as monotherapy with 10 mg / kg QD of sotarasib or 30 mg / kg QD of the compound of formula I. One additional group received combination therapy with 30 mg / kg QD of the compound of formula I and 100 mg / kg QD of sotarasib.

[0251] result Figure 16A shows graphs of tumor volume over treatment periods for regimens of compound I alone (30 mg / kg QD), sotorasib alone (10 mg / kg QD), and a combination of compound I (30 mg / kg QD) and sotorasib (100 mg / kg QD) in the KRAS G12C mutant NSCLC CDX model NCI-H2122. Figure 16B shows graphs of tumor volume versus treatment duration for KEAP1 mutant and KRAS G12C mutant NSCLC CDX model NCI-H2122 tumor xenograft models treated with vehicle (black circles, line 1), sotorasib alone (100 mg / kg QD, black circles, line 2), compound of formula I alone (10 mg / kg / dose BID, black circles, line 3), and a combination of compound of formula I (10 mg / kg / dose BID) and sotorasib (100 mg / kg QD, black circles, line 4).

[0252] knot As illustrated in Figures 16A and 16B, the combination of the compound of formula I with sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant NSCLC CDX model NCI-H2122.

[0253] Example 11: In vivo studies of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model CRC022. material and method Vehicle / control: 100 mM acetic acid in deionized water was prepared by adjusting the pH to 4.8-5.0 and stored under ambient conditions throughout 28 days of administration to mice. Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination agent sotorasib was prepared in a vehicle of 50% w / w polyethylene glycol 400 (PEG400) and 50% w / w propylene glycol (PG) and stored at 2-8°C.

[0254] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were 6–8 weeks old at the time of transplantation. All procedures related to animal handling, care, and treatment in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Institutional Animal Care and Use Committee (IACUC). The animal facilities and programs operated under the standards of the Guideforthe Care and Use of Laboratory Animals (NRC, 2011) and were accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0255] Preparation of PDX The CRC022 PDX model was established for preclinical efficacy studies at GenenDesign (Shanghai, China). This PDX model was derived from a 49-year-old female Chinese CRC patient. The KRAS G12C mutation in the PDX model CRC022 was confirmed by whole exome sequencing and PCR sequencing. Tumor fragments taken from the PDX model were subcutaneously transplanted into the right flank of female Balb / c nude mice. Mice were anesthetized with isoflurane and anesthesia was maintained throughout the transplantation procedure. The mouse skin was cleaned with appropriate surgical scrub and alcohol on the right flank. A small skin incision was made using the sharp end of a trocar, and a 1.5 cm subcutaneous pocket along the right chest wall was formed by blunt incision with a 10-12 g trocar needle stylet. Tumor fragments (15-30 mm) 3 The trocar needle was inserted and advanced into a subcutaneous pocket in the right flank. The trocar incision was closed with sutures or wound clips and removed one week after closure. The tumor size was approximately 200 mm in volume.3 When the average value was reached, the mice with tumors were randomly divided into test groups of 8 mice each. The randomization day is denoted as treatment day 0.

[0256] treatment Treatment was initiated on the day of randomization. The treatment initiation day was designated as treatment day 0. Mice were administered orally with a vehicle control solution, monotherapy with the compound of formula I at 30 mg / kg QD, and monotherapy with sotorasib at 100 mg / kg QD. One additional group received combination treatment with the compound of formula I at 30 mg / kg QD and sotorasib at 100 mg / kg QD. The administration volume was 5 mL / kg for each compound. Sotorasib was administered 1 hour after the administration of the compound of formula I QD in the combination group. This study was concluded on treatment day 28 as defined in the study protocol.

[0257] result Figure 17 shows graphs of tumor volume over treatment periods for regimens of compound I alone, sotorasib alone, and a combination of compound I and sotorasib in the KRAS G12C mutant CRC PDX model CRC022. No significant changes in body weight were observed in either the control or treatment groups.

[0258] knot As shown in Figure 17, the combination of the compound of formula I and sotorasib demonstrated superior tumor growth inhibition compared to each monotherapy in the KRAS G12C mutant CRC PDX model CRC022.

[0259] While the embodiments described above are explained in some detail by illustration and examples for the purpose of clarifying understanding, those skilled in the art will understand that some changes and modifications can be practiced within the scope of the appended claims. Furthermore, each reference provided herein is incorporated by reference to the same extent that each individual reference is incorporated by reference. In the event of any conflict between this application and the references provided herein, this application shall prevail.

Claims

1. 1. A composition for use in a method of treating a subject having cancer, the composition comprising a compound of Formula I: 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, wherein the method comprises administering to the subject the composition in combination with a KRAS inhibitor.

2. 2. The composition of claim 1, wherein the cancer comprises a KRAS G12, G13, H95, Q61, R68 or Y96 mutation.

3. The composition of claim 1, wherein the cancer has a G12C KRAS mutation.

4. The composition of claim 1, wherein the KRAS inhibitor is selected from the group consisting of AMG510 (sotorasib), MRTX849 (adagrasib), ARS-3248, GDC-6036, BI 1701963, tipifarnib, and BBP-454.

5. The composition described in claim 1, wherein the method includes administering a third MAPK pathway inhibitor.

6. The composition of claim 1 , wherein the administration is oral.

7. The composition of claim 1, wherein the dosage of said compound of formula I is in the range of 20 mg to 400 mg per day.

8. The composition of claim 1, wherein the dosage of the compound of formula I is in the range of 10 mg to 140 mg per day.

9. The composition of claim 1, wherein the dosage of the compound of formula I is 10 mg to 300 mg per day, 10 mg to 100 mg per day, or 5 mg to 50 mg per day.

10. The composition of claim 1, wherein the dosage of the compound of formula I is about 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 110 mg / day, 120 mg / day, 130 mg / day, 140 mg / day, 150 mg / day, 160 mg / day, 170 mg / day, 180 mg / day, 190 mg / day or 200 mg / day.

11. 10. The method of claim 1, wherein the compound of formula I, or a pharma- ceutically acceptable salt thereof, is administered once or twice daily.

12. 2. The composition of claim 1, wherein the cancer is selected from lung cancer, gastric cancer, liver cancer, colon cancer, kidney cancer, breast cancer, pancreatic cancer, pancreatic ductal adenocarcinoma (PDAC), juvenile myelomonocytic leukemia, neuroblastoma, melanoma, and acute myeloid leukemia.

13. The composition described in claim 1, wherein the cancer is head and neck cancer.

14. The composition of claim 1, wherein the cancer is rectal cancer.

15. (i) Formula I: 【Chemistry 2】 or a pharma- ceutically acceptable salt thereof, (ii) A kit comprising a KRAS inhibitor.

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