Combination therapy with substituted pyrimidin-4 (3h)-one and sotorasib

Abstract

The present disclosure provides a method of treating cancer in a subject. The method comprises administering to the subject a) a therapeutically effective amount of a compound of formula (I); and b) a therapeutically effective amount of sotoprasib, wherein the compound of formula (I) is represented by the following formula: # imgabs0 # or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformation isomer, tautomer, or a combination thereof. In particular, the present disclosure provides a method of treating a solid tumor (e.g., a tumor tumor) in a subject using a therapeutically effective amount of a compound of formula (10b) (i.e., 6-((3S, 4S)-4-amino-3-methyl-2-oxa-8-azaspiro [4.5] dec-8-yl)-3-(Ra)-(2, 3-dichlorophenyl)-2, 5-dimethylpyrimidin-4 (3H)-one) in combination with sotoprasib. The present invention relates to a method for treating a subject (e.g., advanced or metastatic non-small cell lung cancer) wherein the subject has one or more mutations of KRAS, such as KRAS G12C.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 250,883, filed on September 30, 2021, the entire contents of which are incorporated herein by reference for all purposes.

[0003] STATEMENT REGARDING RIGHTS TO INVENTS RESULTING FROM FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

[0004] not applicable.

[0005] Appendix to a "Sequence Listing", table or computer program listing submitted on a compact disc

[0006] not applicable. Background Art

[0007] The mitogen-activated protein kinase (MAPK) signaling pathway plays a key role in regulating a variety of cellular activities, including cell proliferation, survival, differentiation, and motility. The MAPK signaling pathway is mediated by receptor tyrosine kinases (RTKs). More than one-third of all malignancies present with dysregulation of the MAPK pathway. The classical MAPK pathway consists of Ras (a family of related proteins expressed in all animal cell lineages and organs), Raf (a family of three serine / threonine-specific protein kinases associated with retroviral oncogenes), MEK (mitogen-activated protein kinase kinase), and ERK (extracellular signal-regulated kinase), which sequentially transmit proliferative signals generated by cell surface receptors to the nucleus via cytoplasmic signal transduction. RTKs and components of the MAPK pathway, such as RAS and RAF, are frequently activated by mutations in human cancers, resulting in constitutive pathway activation. Several RTK and MAPK pathway inhibitors, including epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), and MAPK inhibitors, have been approved for the treatment of solid tumors where activation of these pathways is an oncogenic driver.

[0008] KRAS (Kirsten rat sarcoma 2 viral oncogene homolog) is part of the RAS / MAPK pathway. KRAS is one of the most prevalent oncogenes in a variety of human cancers. Despite compelling evidence that oncogenic KRAS drives tumorigenesis, efforts to target mutant KRAS have stalled for years. Recently, progress has been made in the development of treatments for cancers characterized by KRAS with a G12C mutation, which is present in approximately 15% of lung adenocarcinomas and 0%-8% of other cancers. Results from early clinical trials suggest that many cancer patients in this subgroup could benefit significantly from these new therapies. The first KRAS G12C inhibitor, sotolagasib, has been granted accelerated approval by the U.S. Food and Drug Administration (FDA) for the treatment of adult patients with locally advanced or metastatic NSCLC with a KRASG12C mutation, as determined by an FDA-approved test, who have previously received at least one systemic therapy. Another KRAS G12C inhibitor, adagracib, has been granted FDA Breakthrough Therapy designation for the same indication. Other KRAS G12C inhibitors are also under investigation.

[0009] Despite the progress made by RTK inhibitors and MAPK pathway inhibitors, resistance to these pathway inhibitors has been observed in both clinical and nonclinical studies. Such resistance may be driven by activation or upregulation of other components of the pathway. Therefore, monotherapy containing RTK and MAPK pathway inhibitors may not be sufficient to treat tumors that have developed or are likely to develop resistance to approved pathway inhibitors.

[0010] Non-receptor protein-tyrosine phosphatase type 11 (PTPN11, also known as Src homology-2 phosphatase (SHP2)) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene. SHP2 plays a key role in the RTK-mediated MAPK signal transduction pathway. The PTP contains two tandem Src homology-2 (SH2) domains, which serve as the phosphotyrosine binding domain, the catalytic domain, and the C-terminal tail. In the basal state, the protein usually exists in an inactive, self-inhibited conformation, in which the N-terminal SH2 domain blocks the active site. When stimulated by cytokine-mediated signal transduction and growth factor binding of phosphorylated proteins to the SH2 domain, autoinhibition is relieved, making the active site available for dephosphorylation of PTPN11 substrates (MG Mohl, BG Neel, Curr. Opin. Genetics Dev. 2007, 17, 23-30. KS Grossmann, Adv. Cancer Res. 2010, 106, 53-89. WQ Huang et al., Curr. Cancer Drug Targets 2014, 14, 567-588. C. Gordon et al., Cancer Metastasis Rev. 2008, 27, 179-192.).

[0011] Germline and somatic mutations in PTPN11 have been reported in several human diseases, resulting in gain of function of catalytic activity, including Noonan syndrome and Leopard syndrome; and a variety of cancers, such as juvenile myelomonocytic leukemia, neuroblastoma, myelodysplastic syndrome, B-cell acute lymphoblastic leukemia / lymphoma, melanoma, acute myeloid leukemia, and breast, lung and colon cancer (MG Mohl, BG Neel, Curr. Opin. Genetics Dev. 2007, 17, 23-30). Recent studies have shown that single PTPN11 mutations can induce Noonan syndrome, JMML-like myeloproliferative disease and acute leukemia in mice. These mutations disrupt the autoinhibition between the N-SH2 domain and the catalytic site, allowing the substrate to constitutively enter the catalytic site of the enzyme (E. Darian et al., Proteins, 2011, 79, 1573-1588. ZH Yu et al., JBC, 2013, 288, 10472, W Qiu et al., BMC Struct. Biol. 2014, 14, 10).

[0012] PTPN11 is widely expressed in most tissues and plays a regulatory role in various cell signaling events that are essential for a variety of cellular functions including proliferation, differentiation, cell cycle maintenance, epithelial-mesenchymal transition (EMT), mitogenic activation, metabolic control, transcriptional regulation, and cell migration through multiple signaling pathways including the Ras-MAPK, JAK-STAT, or P13K-AKT pathways (Tajan, M. et al., Eur. J. Medical Genetics, 2015, 58, 509-525. Prahallad, A. et al., Cell Reports, 2015, 12, 1978-1985).

[0013] In addition, increasing evidence suggests that PTPN11 / SHP2 is associated with immune escape during tumorigenesis. Therefore, SHP2 inhibitors can stimulate the immune response of cancer patients (Cancer Res. 2015 Feb 1; 75(3): 508-18. T Yokosuka T, J Exp Med. 2012, 209(6), 1201. S Amarnath Sci Transl Med. 2011, 3, 111ra120. T Okazaki, PNAS 2001, 98: 24, 13866-71).

[0014] Substituted pyrimidine-4(3H)-one compounds refer to a class of compounds having inhibitory activity against PTPN11 / SHP2, as disclosed in International Patent Application No. PCT / US2019 / 045903 filed on August 9, 2019, and are represented by the following formula:

[0015]

[0016] or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformer, tautomer or combination thereof, wherein subscripts a and b, Y1, Y2, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 and R 13 As provided in PCT / US2019 / 045903, the entire contents of which are incorporated herein for all purposes. In particular, the substituted pyrimidin-4(3H)-one compounds are represented by formula (I):

[0017]

[0018] Or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformational isomer, tautomer or combination thereof. In certain embodiments, the compound of formula (I) is compound (10b), represented by the following formula:

[0019]

[0020] Its name is 6-((3S,4S)-4-amino-3-methyl-2-oxa-8-oxaspiro[4.5]dec-8-yl)-3-(Ra)-(2,3-dichlorophenyl)-2,5-dimethylpyrimidin-4(3H)-one.

[0021] The compound of formula (I) (particularly the compound of formula (10b)) is a potent, selective, orally active allosteric inhibitor of Src homology-2 phosphatase (SHP2) (also known as protein tyrosine phosphatase non-receptor type 11 (PTPN11)), which is a tyrosine phosphatase that plays a key role in the receptor tyrosine kinase (RTK)-mediated mitogen-activated protein kinase (MAPK) signal transduction pathway (Matozaki, 2009). The key components of the MAPK pathway include small GTPase RAS, serine / threonine-protein kinase RAF, mitogen-activated protein kinase (MEK) and extracellular signal-activated kinase (ERK). In cells, SHP2 binds to phosphorylated tyrosine residues in the intracellular domain of RTK (such as epidermal growth factor receptor (EGFR)), thereby leading to the activation of the downstream MAPK signal transduction pathway.

[0022] Because cancers may be resistant or develop resistance to RTK and MAPK pathway inhibitors, including KRAS G12C inhibitors, there remains a need for effective and safe therapeutic agents for treating cancer, including agents that can be used in combination. Summary of the invention

[0023] The present disclosure provides methods for treating various cancers, comprising administering a KRAS G12C inhibitor (e.g., sotolacib, also known as AMG510) and a PTPN11 inhibitor (e.g., a compound of formula (I) or (10b)). The combination therapies provided herein are used to treat cancer, including lung cancer (e.g., non-small cell lung cancer).

[0024] In one aspect, the present disclosure provides a method of treating cancer (e.g., advanced or metastatic non-small cell lung cancer) in a subject in need thereof, the method comprising administering to the subject:

[0025] a) a therapeutically effective amount of a compound represented by formula (I):

[0026]

[0027] or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformer, tautomer, or combination thereof; and

[0028] b) a therapeutically effective amount of sotolacib.

[0029] In another aspect, the present disclosure provides a pharmaceutical composition for treating cancer in a subject, the composition comprising:

[0030] a) a therapeutically effective amount of a compound represented by formula (I); and

[0031] b) a therapeutically effective amount of sotolacib,

[0032] and a pharmaceutically acceptable carrier or excipient,

[0033] wherein the compound of formula (I) is as defined and described herein.

[0034] In another aspect, the present disclosure provides a kit for treating cancer in a subject, the kit comprising:

[0035] a) a therapeutically effective amount of a compound represented by formula (I); and

[0036] b) a therapeutically effective amount of sotolacib,

[0037] and instructions for effective administration,

[0038] wherein the compound of formula (I) is as defined and described herein.

[0039] In some embodiments, the compound of formula (I) is represented by formula (10b):

[0040]

[0041] Its name is 6-((3S,4S)-4-amino-3-methyl-2-oxa-8-oxaspiro[4.5]dec-8-yl)-3-(Ra)-(2,3-dichlorophenyl)-2,5-dimethylpyrimidin-4(3H)-one. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figures 1A-1E It was shown that the combination of a compound of formula (10b) and compound A (i.e., 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one) inhibited the growth of NCI-H358 subcutaneous tumors at a tolerated dose in a mouse model. Figure 1A , Figure 1B and Figure 1D Tumor volumes monitored biweekly by caliper are shown. Figure 1C and Figure 1E The recorded body weights are shown. Data are expressed as mean ± SEM. Figure 1B-1E , N = 8 mice / group and Figure 1A , N = 10 mice / group

[0043] Figure 2A and Figure 2B Shown are changes in tumor volume and body weight of mice bearing tumors derived from the NCI-H358 subcutaneous cell line after treatment with the compound of formula (10b) alone or in combination with sotolacib (AMG 510).

[0044] Figure 3A and Figure 3B Shown are changes in tumor volume and body weight of mice bearing tumors derived from the NCI-H2122 subcutaneous cell line after treatment with the compound of formula (10b) alone or in combination with sotolacib (AMG 510).

[0045] Figures 4A-4D showed that in NCI-H358 (KRAS G12C ) model, in vitro activity data of the compound of formula (10b) and sotolacib (AMG510). Figure 4A : 3D activity at various concentrations of formula (10b). Figure 4B : Biss energy score for the combination of AMG 510 and formula (10b). Figure 4C : Fold change in DUSP6 levels. Figure 4D : Fold change of SPRY4 level. Figure 4C-4D As shown in , the combination of AMG 510 and formula (10b) inhibited the levels of DUSP6 and SPRY4 relative to either agent alone.

[0046] Figure 5 A Phase 1A / 1B study of the combination of Formula (10b) and sotolacib in patients with solid tumors harboring a KRAS G12C mutation is shown. The study design includes Phase 1a dose escalation and Phase 1b dose expansion / optimization.

[0047] Figure 6 A flow chart showing a trial conducted using the BOIN design. Abbreviations: BOIN = Bayesian optimal interval design; DLT = dose-limiting toxicity; MTD = maximum tolerated dose. Notes: λe = 19.7% and λd = 29.8%. In practice, each cohort of 6 patients was dose-escalated if the DLT rate was ≤ 1 / 6 and dose-reduced if the DLT rate was ≥ 2 / 6. DETAILED DESCRIPTION

[0048] I. Overview

[0049] The present disclosure provides a method for combined therapy of cancer (e.g., solid tumor) in a subject. The method includes administering to the subject a) a therapeutically effective amount of a compound of formula (I) (such as a PTPN11 inhibitor); and b) a therapeutically effective amount of sotolacib, wherein the compound of formula (I) is as defined and described herein. In particular, cancer is characterized by KRAS mutations, such as mutations other than Q61X mutations, for example, KRAS G12C mutations. In some cases, cancer is a solid tumor, such as advanced or metastatic non-small cell lung cancer (NSCLC). Also provided is a pharmaceutical composition and a kit thereof for treating cancer in a subject.

[0050] II. Definitions

[0051] As used herein, the following terms have the indicated meanings.

[0052] "Include," "including," and "having" and their derivatives are used interchangeably herein as comprehensive, open-ended terms. For example, the use of "include," "comprising," and "having" means that whatever elements are included, having, or comprising are not the only elements included by the subject of the clause containing the verb.

[0053] When a range of values ​​is disclosed, and the expression "from n1 ... to n2" or "between n1 ... and n2" is used, where n1 and n2 are numbers, then unless otherwise specified, the expression is intended to include the numbers themselves and the range between them. The range can be an integer or continuous (including the endpoint values) between the endpoint values. For example, the range "2 to 6 carbons" is intended to include two, three, four, five, and six carbons, because carbon appears in integer units. In contrast, the range "1 to 3 μM (micromolar)" is intended to include 1 μM, 3 μM, and all significant figures between the two (e.g., 1.255 μM, 2.1 μM, 2.9999 μM, etc.).

[0054] "Salt" refers to an acid salt or base salt of a compound of the present disclosure. Illustrative examples of pharmaceutically acceptable acid addition salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, etc.) salts and organic acid (acetic acid, propionic acid, glutamic acid, citric acid, etc.) salts. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts or similar salts. It should be understood that pharmaceutically acceptable salts are non-toxic. Additional information about suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, Pa., 1985, which is incorporated herein by reference.

[0055] "Solvate" refers to a compound provided herein, or a salt thereof, that further includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces.

[0056] "Hydrate" refers to a compound complexed with a water molecule. The compounds of the present disclosure may be complexed with 1 / 2 water molecule or 1 to 10 water molecules.

[0057] There are asymmetric centers in the compounds disclosed herein. These centers are represented by the symbols "R" or "S", depending on the configuration of the substituents around the chiral carbon atom. It should be understood that the present invention encompasses all stereochemical isomeric forms, including diastereoisomers, enantiomers and epimers, as well as d-isomers and 1-isomers and mixtures thereof. Each stereoisomer of a compound can be synthesized from commercially available starting materials containing a chiral center, or by preparing a mixture of enantiomeric products, which are then separated (e.g., converted into a mixture of diastereoisomers and then separated) or by recrystallization, chromatography, direct separation of enantiomers on a chiral chromatographic column, or any other suitable method known in the art. Starting compounds with specific stereochemistry can be commercially available, or can be prepared and resolved by techniques known in the art. In addition, the compounds disclosed herein can exist in the form of geometric isomers. The present disclosure includes all cis, trans, syn, anti, E-form and Z-form isomers, and appropriate mixtures thereof. In addition, the compounds may exist in the form of tautomers; the present disclosure provides all tautomers. In addition, the compounds disclosed herein may exist in unsolvated and solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc. In general, the solvated forms are considered equivalent to the unsolvated forms.

[0058] "Tautomer" used herein alone or in combination refers to one of two or more isomers that rapidly interconvert. Typically, this interconversion is fast enough, so in the absence of another tautomer, a single tautomer will not be separated. The ratio of the amount of tautomers can depend on solvent composition, ionic strength and pH and other solution parameters. In a specific solution and the microenvironment of the biomolecule binding site in the solution, the ratio of the amount of tautomers may be different. Examples of tautomers include keto / enol, enamine / imine and lactam / lactim tautomers. Other examples of tautomers also include 2-hydroxypyridine / 2 (1H)-pyridone and 2-aminopyridine / 2 (1H)-iminopyridone tautomers.

[0059] Conformation isomers exist in the compounds disclosed herein. Wherein, when R1 is aryl or heteroaryl:

[0060]

[0061] The aryl or heteroaryl groups are in different conformations relative to the pyrimidinone moiety as shown below:

[0062]

[0063] These forms are represented by the symbol "S a ” or “R a " indicates that it depends on the conformation of the aryl or heteroaryl group relative to the pyrimidinone portion. "S a ” and “R a ", the entire contents of which are incorporated herein for all purposes. The compound of formula (10b) is essentially "R a "form.

[0064] "Pharmaceutically acceptable" refers to those compounds (salts, hydrates, solvates, stereoisomers, conformers, tautomers, etc.) that are suitable for contact with patient tissues without excessive toxicity, irritation and allergic response, commensurate with a reasonable benefit / risk ratio, and effective for its intended use. As defined and described herein, the compounds disclosed herein may exist as pharmaceutically acceptable salts.

[0065] The term "combination therapy" refers to the administration of two or more therapeutic agents to treat the conditions or disorders described in the present disclosure. Such administration encompasses the co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule with a fixed ratio of active ingredients or in a plurality of separate capsules of each active ingredient. In addition, such administration also encompasses the use of each type of therapeutic agent in a sequential manner. In either case, the above treatment regimen will provide a beneficial effect of the drug combination in treating the conditions or disorders described herein.

[0066] As used herein, a "PTPN11 inhibitor" refers to an inhibitor that exhibits an IC relative to PTPN11 activity as measured by a PTPN11 assay generally described in International Patent Application No. PCT / US2019 / 045903 (e.g., the enzymatic activity of the recombinant human PTPN11 protein of Example 21). 50 A compound that is no greater than about 100 micromolar (μM), more usually no greater than about 50 μM. 50 ” is the concentration of an inhibitor that reduces the activity of an enzyme (e.g., PTPN11) to half-maximal levels. In certain embodiments, the compounds disclosed in PCT / US2019 / 045903 exhibit an IC of no more than about 10 μM for PTPN11 inhibition 50 In a further embodiment, the compound exhibits an IC of no more than about 1 μM for PTPN11 inhibition 50 In yet further embodiments, the compound exhibits an IC of no more than about 200 nM for PTPN11 inhibition 50 In yet further embodiments, the compound exhibits an IC of no more than about 100 nM for PTPN11 inhibition 50 and in yet further embodiments, the compound exhibits an IC of no more than about 50 nM for PTPN11 inhibition 50 , as measured in the PTPN11 assay described herein. In certain embodiments, the compound of formula (I) or (10b) exhibits an IC of no more than 50 nM for PTPN11 (e.g., PTPN11-E76K mutant enzyme) inhibition. 50 .

[0067] "Therapeutically effective amount" refers to the amount of a compound or pharmaceutical composition that can be used to treat or improve an identified disease or condition, or can be used to show a detectable therapeutic or inhibitory effect. The exact amount will depend on the purpose of the treatment, and can be determined by a person skilled in the art using known techniques (see, e.g., 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).

[0068] "Treat," "treating," and "treatment" are any markers of success in treating or ameliorating an injury, pathology, or condition, including any objective or subjective parameter, such as alleviating, relieving, reducing symptoms, or making a patient more tolerant to an injury, pathology, or condition; slowing the rate of degeneration or decline; reducing worsening of the final endpoint of degeneration; and / or improving the patient's physical or mental health. Treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neuropsychiatric examination, and / or psychiatric evaluation.

[0069] "Administering" refers to therapeutically providing a compound or a form thereof to a subject, such as by oral administration or intravenous administration.

[0070] "Patient" or "subject" refers to a living organism suffering from or susceptible to a disease or condition that can be treated by administering a pharmaceutical composition of the present disclosure. Non-limiting examples include humans, non-human primates (e.g., monkeys), goats, pigs, sheep, cattle, deer, horses, cows, rats, mice, rabbits, hamsters, guinea pigs, cats, dogs, and other non-mammals. In some embodiments, the subject is a human. In some embodiments, the subject is an adult (e.g., at least 18 years old).

[0071] As used herein, "composition" is intended to encompass a product of the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts. "Pharmaceutically acceptable" is intended to mean the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0072] "Pharmaceutically acceptable excipient" refers to a substance that facilitates administration of an active agent to a subject and facilitates absorption by the subject. Pharmaceutical excipients that may be used in the present disclosure include, but are not limited to, binders, fillers, disintegrants, lubricants, coating agents, sweeteners, flavoring agents, and coloring agents. Other pharmaceutical excipients may also be used in the present disclosure.

[0073] "Tablet" refers to a solid pharmaceutical preparation with and without coating. The term "tablet" also refers to a tablet with one, two, three or even more layers, wherein the various aforementioned types of tablets may be without or with one or more layers of coating. In some embodiments, the tablets of the present disclosure may be prepared by roller compaction or other suitable means known in the art. The term "tablet" also includes mini tablets, melting tablets, chewable tablets, effervescent tablets and orally disintegrating tablets. The tablet comprises a compound of formula (I) or (10b) and one or more pharmaceutically acceptable excipients (e.g., fillers, binders, glidants, disintegrants, surfactants, binders, lubricants, etc.). Optionally, a coating agent may also be included. In order to calculate the weight percentage of the tablet formulation, the amount of the coating agent is not included in the calculation. That is, the weight percentage reported herein is for uncoated tablets.

[0074] Unless otherwise specified, the content of the compound of formula (I) or (10b) in, for example, a tablet formulation is calculated based on the standardized weight of the compound of formula (I) or (10b) on a salt-free and water-free basis. That is, the salt and / or water content in the compound of formula (I) or (10b) is not included in the calculation.

[0075] As used herein, "KRAS G12C inhibitor" refers to a compound that targets, reduces or inhibits the synthesis or biological activity of KRAS (Kirsten rat sarcoma 2 viral oncogene homolog) by selectively modifying the mutant cysteine ​​12 in the KRAS of the G12C mutation. KRAS G12C inhibitors can at least partially inhibit KRAS G12C. KRAS G12C inhibitors can be selective KRAS G12C inhibitors (e.g., having greater selectivity for KRAS with G12C mutations relative to KRAS with another mutation (such as G12D mutations)). In those cases, selective KRAS G12C inhibitors can have high potency for KRAS G12C, and can have low affinity for other KRAS mutations. KRAS G12C inhibitors can be covalent inhibitors (e.g., capable of covalently modifying cysteine ​​12). KRAS G12C inhibitors can be non-covalent inhibitors. KRASG12C inhibitors can bind to the inactive ("GDP") form of KRAS. KRAS G12C inhibitors can bind to the active ("GTP") form of KRAS. KRAS G12C inhibitors can bind to both the inactive ("GDP") and active ("GTP") forms of KRAS. Examples of KRAS G12C inhibitors include sotolacib (AMG 510), adagracib (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI1823911, MK-1084, LY3537982, and LY3499446.

[0076] "KRAS-positive cancer" refers to a cancer in which the KRAS gene has been rearranged, mutated, or amplified. "KRAS G12C-positive cancer" refers to a cancer in which the KRAS G12C gene has been rearranged, mutated, or amplified.

[0077] "Cancer resistant to KRAS inhibitors" and / or "KRAS-positive cancer resistant to KRAS inhibitors" refers to a cancer or tumor that has failed to respond favorably to treatment with a previous KRAS inhibitor, or alternatively has recurred or relapsed after a favorable response to a KRAS inhibitor. "Cancer resistant to KRAS G12C inhibitors" and / or "KRAS G12C-positive cancer resistant to KRAS G12C inhibitors" refers to a cancer or tumor that has failed to respond favorably to treatment with a previous KRASG12C inhibitor, or alternatively has recurred or relapsed after a favorable response to a KRAS G12C inhibitor.

[0078] As used herein, "combined therapeutically effective amount" means an amount of the therapeutic agents that, when administered separately (in a time-sequentially staggered manner, particularly in a sequentially specific manner) to a warm-blooded animal, especially a human being, shows an (additive, but preferably synergistic) interaction (combined therapeutic effect). Whether this is the case can be determined by following blood levels, indicating that at least at certain time intervals, both compounds are present in the blood of the person being treated.

[0079] As used herein, "synergistic effect" refers to at least two therapeutic agents: a PTPN11 inhibitor as defined herein; and a KRAS G12C inhibitor as defined herein that has an effect greater than the simple addition of the effects of the various drugs administered alone. The effect can be, for example, slowing the progression of symptoms of a proliferative disease (such as cancer, particularly lung cancer (e.g., non-small cell lung cancer)) or its symptoms. Similarly, a "synergistically effective amount" refers to the amount required to obtain a synergistic effect.

[0080] "A", "an" or "a(n)" when used herein to refer to a group of substituents or "substituents" means at least one. For example, when a compound is substituted with "an" alkyl or aryl group, the compound is substituted with at least one alkyl group and / or at least one aryl group, wherein each alkyl group and / or aryl group is optionally different. In another example, when a compound is substituted with "a" substituent group, the compound is substituted with at least one substituent group, wherein each substituent group is optionally different.

[0081] III. Combination Therapy

[0082] In one aspect, the present disclosure provides a method of treating cancer in a subject. The method comprises administering to the subject:

[0083] a) a therapeutically effective amount of a compound represented by formula (I):

[0084]

[0085] or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformer, tautomer, or combination thereof; and

[0086] b) a therapeutically effective amount of sotolacib (AMG 510).

[0087] III-1: Compound of formula (I)

[0088] The compounds of formula (I) may be in the form of pharmaceutically acceptable salts or neutral forms, each of which may optionally be in the form of solvates or hydrates.

[0089] In some embodiments, the compound of formula (I) is in the form of a pharmaceutically acceptable salt. In some embodiments, the pharmaceutically acceptable acid addition salt of the compound of formula (I) is represented by formula (Ia):

[0090]

[0091] wherein HX is a pharmaceutically acceptable acid addition product.

[0092] Examples of acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and the like, and salts derived from organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, linoleic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like.

[0093] In some embodiments, the compound of formula (I) is in a neutral form.

[0094] In some embodiments, the compound of formula (I) has a substantial portion of 6-((3,S,4S)-4-amino-3-methyl-2-oxa-8-oxaspiro[4.5]dec-8-yl) having a stereochemistry as shown in formula (10b):

[0095]

[0096] In some embodiments, the compound of formula (I) is substantially R a Conformation, as shown in formula (10b):

[0097]

[0098] In some embodiments, the compound of formula (I) is represented by formula (10b):

[0099]

[0100] Its name is 6-((3S,4S)-4-amino-3-methyl-2-oxa-8-oxaspiro[4.5]dec-8-yl)-3-(Ra)-(2,3-dichlorophenyl)-2,5-dimethylpyrimidin-4(3H)-one.

[0101] In some embodiments, the compound of formula (I) or (10b) is in a neutral form.

[0102] In some embodiments, the compound of formula (I) includes one or more corresponding enantiomers, diastereomers and / or conformers of the compound of formula (10b), as represented by the following formulae:

[0103]

[0104] In some embodiments, the compound of formula (10b) has a purity of at least about 95 area % as measured by chiral high performance liquid chromatography (HPLC). In some embodiments, the compound of formula (10b) has a purity of from about 95 area % to about 99 area %, from about 96 area % to about 99 area %, from about 97 area % to about 99 area % or from about 98 area % to about 99 area % as measured by chiral high performance liquid chromatography (HPLC). In some embodiments, the compound of formula (10b) has a purity of from about 98 area % to about 99 area %.

[0105] In some embodiments, the compound of formula (I) includes one or more corresponding enantiomers, diastereomers and / or conformers of the compound of formula (10b), as shown above; and the sum of the one or more isomers does not exceed about 5 area % as determined by chiral high performance liquid chromatography (HPLC).

[0106] In some embodiments, the corresponding enantiomers, diastereomers and / or conformers of the compound of formula (10b) are present in the compound of formula (I) and meet the following qualifying criteria: enantiomers (3R, 4R, S a )≤0.5 area %; diastereoisomers (3R, 4S, R a )≤1.2 area %; diastereoisomers (3S, 4R, S a )≤0.5 area %; diastereoisomers (3R, 4R, R a )≤0.5 area %; diastereoisomers (3S, 4S, S a )≤0.5 area %; diastereoisomers (3S, 4R, R a )≤0.5 area %; and diastereomers (3R, 4S, S a )≤0.5 area %, each of which is determined by chiral high performance liquid chromatography (HPLC). In some embodiments, the compound of formula (10b) has a purity of at least about 95 area %, wherein the enantiomers (3R, 4R, S a )<0.5 area %; diastereoisomers (3R, 4S, R a )<1.2 area %; diastereoisomers (3S, 4R, S a )<0.5 area %; diastereoisomers (3R, 4R, R a )<0.5 area %; diastereoisomers (3S, 4S, S a )<0.5 area %; diastereomers (3S, 4R, R a)<0.5 area %; and diastereomers (3R, 4S, S a ) <0.5 area %, each of which is determined by chiral high performance liquid chromatography (HPLC). In some embodiments, the compound of formula (10b) has a purity from about 95 area % to about 99 area %, from about 96 area % to about 99 area %, from about 97 area % to about 99 area % or from about 98 area % to about 99 area %, wherein the enantiomers (3R, 4R, S a )<0.5 area %; diastereoisomers (3R, 4S, R a )<1.2 area %; diastereoisomers (3S, 4R, S a )<0.5 area %; diastereoisomers (3R, 4R, R a )<0.5 area %; diastereoisomers (3S, 4S, S a )<0.5 area %; diastereomers (3S, 4R, R a )<0.5 area %; and diastereomers (3R, 4S, S a ) < 0.5 area %, each of which is determined by chiral high performance liquid chromatography (HPLC). In some embodiments, the compound of formula (10b) has a purity of from about 98 area % to about 99 area %, wherein the enantiomers (3R, 4R, S a ) not detected; diastereomers (3R, 4S, R a ) is about 0.86 area %; diastereomers (3S, 4R, S a ) not detected; diastereomers (3R, 4R, R a ) is about 0.07 area %; diastereomers (3S, 4S, S a ) not detected; diastereomers (3S, 4R, R a ) not detected; and diastereomers (3R, 4S, S a ) were not detected, each of which was determined by chiral high performance liquid chromatography (HPLC).

[0107] In some embodiments, the compound of any one of Formula (I), Formula (Ia) and Formula (10b) is in the form of a solvate and / or a hydrate.

[0108] III-2: Cancer / Solid Tumors

[0109] The cancer can be any cancer that responds to treatment with a PTPN11 inhibitor and / or a KRAS G12C inhibitor (e.g., sotolacib). In some embodiments, the cancer is caused by and / or is characterized by a KRAS mutation (e.g., a KRAS G12C mutation). In some embodiments, the cancer is characterized by a KRAS mutation other than a Q61X mutation. In some embodiments, the cancer is a KRAS-positive cancer. In some embodiments, the cancer is a KRAS G12C-positive cancer (e.g., a cancer characterized by a G12C mutation in KRAS).

[0110] Cancer can be characterized by solid tumors and / or liquid tumors. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the cancer comprises a liquid tumor.

[0111] In some embodiments, the cancer is lung cancer, colorectal cancer, pancreatic cancer, urothelial carcinoma, gastric cancer, mesothelioma, or a combination thereof. In some embodiments, the cancer is non-small cell lung cancer (NSCLC). In some embodiments, the cancer is NSCLC characterized by a KRAS mutation, such as a KRAS G12C mutation. In certain embodiments, the KRAS protein comprises a G12C mutation. In certain embodiments, the cancer is NSCLC characterized by a G12C mutation in KRAS. In some embodiments, the cancer is NSCLC characterized by a mutation in an epidermal growth factor receptor (EGFR) protein. In some embodiments, the cancer is NSCLC characterized by a mutation in an epidermal growth factor receptor (EGFR) protein. In some embodiments, the cancer is NSCLC characterized by no EGFR or anaplastic lymphoma kinase (ALK) mutation.

[0112] In some embodiments, the cancer is a KRAS G12C positive cancer (e.g., a cancer characterized by a G12C mutation in KRAS). In some embodiments, the KRASG12C positive cancer is non-small cell lung cancer, small intestine cancer, appendix cancer, colorectal cancer, unknown primary cancer, endometrial cancer, mixed cancer types, pancreatic cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell cancer, ovarian cancer, gastrointestinal neuroendocrine cancer, bladder cancer, myelodysplastic / myeloproliferative tumors, head and neck cancer, esophageal gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia or melanoma. In some embodiments, the cancer is small intestine cancer, appendix cancer, endometrial cancer, hepatobiliary cancer, small cell lung cancer, cervical cancer, germ cell tumor, ovarian cancer, gastrointestinal neuroendocrine tumor, bladder cancer, myelodysplastic / myeloproliferative tumors, head and neck cancer, esophageal gastric cancer, soft tissue sarcoma, mesothelioma, thyroid cancer, leukemia or melanoma. In some embodiments, the KRASG12C positive cancer is non-small cell lung cancer, colorectal cancer, pancreatic cancer, appendix cancer, endometrial cancer, unknown primary cancer, ampullary cancer, gastric cancer, small intestine cancer, sinus cancer, bile duct cancer or melanoma. In some embodiments, the cancer is an advanced or metastatic KRASG12C positive solid tumor (e.g., lung cancer, colorectal cancer, pancreatic cancer, urothelial cancer, gastric cancer, mesothelioma or a combination thereof). In some embodiments, the cancer is an advanced or metastatic KRAS G12C positive non-small cell lung cancer (NSCLC). In some embodiments, the cancer is an advanced or metastatic KRAS G12C positive solid tumor, provided that the solid tumor is not non-small cell lung cancer (NSCLC).

[0113] Cancer can also be any cancer that is resistant to treatment with a KRAS G12C inhibitor. In some embodiments, cancer is resistant to a KRAS G12C inhibitor, as defined and described herein. In some embodiments, cancer is resistant to a KRAS G12C inhibitor of an inactive ("GDP") form of an inhibitor of KRAS. In some embodiments, cancer is resistant to a KRAS G12C inhibitor of an active ("GTP") form of an inhibitor of KRAS. In some embodiments, cancer is resistant to a KRAS G12C inhibitor of an inactive ("GDP") and active ("GTP") form of an inhibitor of KRAS.

[0114] In some embodiments, the cancer is characterized by intrinsic and / or acquired resistance to a KRAS G12C inhibitor as defined and described herein. In some embodiments, the cancer is a KRAS G12C-positive cancer that is resistant to a KRAS G12C inhibitor as defined and described herein. In some embodiments, the cancer is a KRAS G12C-positive cancer that is characterized by intrinsic and / or acquired resistance to a KRAS G12C inhibitor as defined and described herein.

[0115] In some embodiments, the cancer is characterized by inherent and / or acquired resistance to another therapy (such as a KRAS modulator, a platinum-based therapy, or a taxane therapy). In some embodiments, the cancer is characterized by inherent and / or acquired resistance to a KRAS G12C inhibitor. In some embodiments, the cancer is characterized by inherent and / or acquired resistance to a KRASG12C inhibitor selected from the following: Sotolasib (AMG510), Adagrasibu (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI1823911, MK-1084, LY3537982, and LY3499446. In some embodiments, the cancer is characterized by inherent and / or acquired resistance to Sotolasib (AMG 510). In some embodiments, the cancer is characterized by intrinsic and / or acquired resistance to adagracib (MRTX-849).

[0116] In some embodiments, the cancer is resistant to a KRAS G12C inhibitor selected from the group consisting of Sotolacib (AMG 510), Adagracib (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI 1823911, MK-1084, LY3537982, and LY3499446. In some embodiments, the cancer is resistant to Sotolacib (AMG 510) or Adagracib (MRTX-849). In some embodiments, the cancer is resistant to Sotolacib (AMG 510). In some embodiments, the cancer is resistant to Adagracib (MRTX-849). In some embodiments, the cancer is a KRAS G12C-positive cancer resistant to a KRAS G12C inhibitor selected from: Sotolasib (AMG 510), Adagraciba (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI 1823911, MK-1084, LY3537982, and LY3499446. In some embodiments, the cancer is a KRAS G12C-positive cancer resistant to Sotolasib (AMG 510) or Adagraciba (MRTX-849). In some embodiments, the cancer is a KRAS G12C-positive cancer resistant to Sotolasib (AMG 510). In some embodiments, the cancer is a KRAS G12C-positive cancer that is resistant to adagraciib (MRTX-849).

[0117] The solid tumor can be any solid tumor that responds to treatment with a PTPN11 inhibitor and a KRAS G12C inhibitor (e.g., sotolacib). In some embodiments, the solid tumor is a tumor in which one or more genes in KRAS have been rearranged, mutated, or amplified, provided that the tumor is not caused by one or more other activating mutations in BRAF V600X, PTPN11 (SHP2), or KRAS Q61X.

[0118] In some embodiments, the solid tumor is advanced or metastatic non-small cell lung cancer (NSCLC) caused by a mutation in KRAS. In some embodiments, the solid tumor is advanced or metastatic non-small cell lung cancer (NSCLC) caused by a mutation in KRAS, provided that the tumor is not caused by one or more other activating mutations in BRAF V600X, PTPN11 (SHP2) or KRAS Q61X. In some embodiments, the solid tumor is a KRAS G12C positive solid tumor. In some embodiments, the solid tumor is advanced or metastatic KRASG12C positive non-small cell lung cancer (NSCLC).

[0119] A solid tumor can also be any tumor that is resistant to treatment with a KRAS G12C inhibitor (e.g., Sotolacib (AMG510), Adagracib (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI1823911, MK-1084, LY3537982, and LY3499446). In some embodiments, the solid tumor is resistant to a KRAS G12C inhibitor. In some embodiments, the solid tumor is characterized by inherent and / or acquired resistance to a KRAS G12C inhibitor as defined and described herein. In some embodiments, the solid tumor is a KRAS G12C-positive solid tumor that is resistant to a KRAS G12C inhibitor. In some embodiments, the solid tumor is characterized by a KRAS G12C positive solid tumor with intrinsic and / or acquired resistance to a KRAS G12C inhibitor. In some embodiments, the solid tumor is resistant to treatment with a KRAS G12C inhibitor selected from the following: Sotolasib (AMG510), Adagracib (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI 1823911, MK-1084, LY3537982, and LY3499446. In some embodiments, the solid tumor is resistant to Sotolasib (AMG 510). In some embodiments, the solid tumor is resistant to Adagracib (MRTX-849). In some embodiments, the solid tumor is a KRAS positive solid tumor resistant to treatment with a KRAS G12C inhibitor selected from the following: sotolasib (AMG 510), adagrasibu (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI1823911, MK-1084, LY3537982 and LY3499446. In some embodiments, the solid tumor is a KRAS positive solid tumor resistant to sotolasib (AMG 510). In some embodiments, the solid tumor is a KRAS positive solid tumor resistant to adagrasibu (MRTX-849).In some embodiments, the solid tumor is a KRAS G12C positive solid tumor resistant to treatment with a KRASG12C inhibitor selected from the following: Sotolasib (AMG 510), Adagracib (MRTX-849), MRTX1257, ARS-853, ARS-1620, JNJ-74699157 (ARS-3248), JDQ443, GDC-6036, JAB-21822, BI 1823911, MK-1084, LY3537982 and LY3499446. In some embodiments, the solid tumor is a KRAS G12C positive solid tumor resistant to Sotolasib (AMG 510). In some embodiments, the solid tumor is a KRAS G12C positive solid tumor resistant to Adagracib (MRTX-849).

[0120] In some embodiments, the cancer (e.g., a solid tumor) is characterized by mutations in one or more MAPK pathways, such as mutations in one or more of the following: KRAS, NRAS, HRAS, CRAF, BRAF, NRAF, MAPK / ERK, MAPKK / MEK, NF1, EGFR, IGFR, PDGFR, VEGFR, FGFR, CCKR, NGFR, EphR, AXLR, KEAP-1, TIE receptor, RYK receptor, DDR receptor, RET receptor, ROS receptor, LTK receptor, ROR receptor, MuSK receptor, or a combination thereof.

[0121] In some embodiments, the cancer or solid tumor is resistant to treatment with inhibitors that target, decrease or inhibit the synthesis, expression or biological activity in the MAPK pathway, such as inhibitors that target one or more of the following: KRAS, NRAS, HRAS, CRAF, BRAF, NRAF, MAPK / ERK, MAPKK / MEK, NF1, EGFR, IGFR, PDGFR, VEGFR, FGFR, CCKR, NGFR, EphR, AXLR, KEAP-1, TIE receptor, RYK receptor, DDR receptor, RET receptor, ROS receptor, LTK receptor, ROR receptor, MuSK receptor, or a combination thereof. In some embodiments, the cancer or solid tumor is characterized by intrinsic and / or acquired resistance to inhibitors that target, reduce or inhibit synthesis, expression or biological activity in the MAPK pathway, such as inhibitors that target one or more of the following: KRAS, NRAS, HRAS, CRAF, BRAF, NRAF, MAPK / ERK, MAPKK / MEK, NF1, IGFR, PDGFR, VEGFR, FGFR, CCKR, NGFR, EphR, AXLR, KEAP-1, TIE receptor, RYK receptor, DDR receptor, RET receptor, ROS receptor, LTK receptor, ROR receptor, MuSK receptor or a combination thereof. Examples of MEK inhibitors include cobimetinib, trametinib, bimetinib, mitraz and selumetinib. Examples of BRAF inhibitors include sorafenib, regorafenib, vemurafenib, vemurafenib, conafenib and dabrafenib. Examples of EGFR inhibitors include erlotinib, cetuximab, panitumumab, vandetanib, afatinib, gefitinib, osimertinib, necituzumab, brigatinib, neratinib, dacomitinib, ivanovumab (JNJ-61186372), mobotinib (TAK-788), BLU-945, valitinib, tarotuximab, pocitinib, and lapatinib.

[0122] In any of the embodiments, standard of care or curative therapy for treating cancer or solid tumors, as described herein, is not applicable.

[0123] III-3: Subject

[0124] In some embodiments, the subject is a human. In some embodiments, the subject is under the care of a practicing physician, such as a physician. In some embodiments, the subject has been diagnosed with cancer. In some embodiments, the subject has relapsed. In some embodiments, the subject has previously entered remission. In some embodiments, the subject has previously undergone, is undergoing, or will undergo a monotherapy course. In some embodiments, the subject has previously undergone, is undergoing, or will undergo radiation therapy. In some embodiments, the subject has previously undergone, is undergoing, or will undergo immunotherapy. In some embodiments, the subject has previously undergone, is undergoing, or will undergo chemotherapy. In some embodiments, the subject has previously undergone, is undergoing, or will undergo platinum-based chemotherapy. In some embodiments, the subject has previously undergone, is undergoing, or will undergo a treatment regimen comprising administering a KRAS modulator (e.g., a KRAS inhibitor). In some embodiments, the subject has previously undergone, is undergoing, or will undergo a treatment regimen comprising administering an anti-PD-1 / PD-L1 inhibitor (e.g., a checkpoint inhibitor).

[0125] The subject may have an advanced (e.g., primary, metastatic, or recurrent) solid tumor with a KRAS G12C mutation (e.g., as described herein), as assessed by molecular diagnostics using an appropriate clinically validated and / or FDA-approved test, and no standard of care or curative therapy is available. In some embodiments, the subject has a KRAS G12C mutation (e.g., as described herein), as assessed by molecular diagnostics using an appropriate clinically validated and / or FDA-approved test at least two (2) years prior to entering treatment as described herein.

[0126] In some embodiments, the subject has a cancer characterized by having one or more mutations in KRAS (e.g., as described herein), provided that the cancer is characterized by a KRAS G12C mutation. In some embodiments, the subject has a cancer characterized by a KRAS G12C mutation and is not characterized by a KRAS Q61X mutation. In some embodiments, the subject has a cancer characterized by a KRAS G12C mutation and other mutations at codon 13 (e.g., G13D, G13A, G13C, G13R, G13S, and G13V mutations). In some embodiments, the subject has a cancer characterized by a KRAS G12C mutation and other mutations at codon 61.

[0127] In some embodiments, the subject has one or more mutations in the MAPK pathway. In some embodiments, the one or more mutations in the MAPK pathway are one or more mutations other than BRAF mutations comprising V600X mutations, provided that the subject also has a KRAS G12C mutation. In some embodiments, the subject has a mutation in one or more MAPK pathways selected from the following: NRAS, HRAS, CRAF, BRAF, NRAF, MAPK / ERK, MAPKK / MEK, NF1, IGFR, PDGFR, VEGFR, FGFR, CCKR, NGFR, EphR, AXLR, TIE receptor, RYK receptor, DDR receptor, RET receptor, ROS receptor, LTK receptor, ROR receptor, MuSK receptor, provided that the subject also has a KRAS G12C mutation. In some embodiments, the subject has a mutation in NRAS. In some embodiments, the subject has a mutation in HRAS. In some embodiments, the subject has a mutation in CRAF. In some embodiments, the subject has a mutation in BRAF (except for V600X mutations). In some embodiments, the subject has a mutation in NRAF. In some embodiments, the subject has a mutation in MAPK / ERK. In some embodiments, the subject has a mutation in MAPKK / MEK. In some embodiments, the subject has a mutation in NF1. In some embodiments, the subject has a mutation in IGFR. In some embodiments, the subject has a mutation in PDGFR. In some embodiments, the subject has a mutation in VEGFR. In some embodiments, the subject has a mutation in FGFR. In some embodiments, the subject has a mutation in CCKR. In some embodiments, the subject has a mutation in NGFR. In some embodiments, the subject has a mutation in EphR. In some embodiments, the subject has a mutation in AXLR. In some embodiments, the subject has a mutation in TIE receptor. In some embodiments, the subject has a mutation in RYK receptor. In some embodiments, the subject has a mutation in DDR receptor. In some embodiments, the subject has a mutation in RET receptor. In some embodiments, the subject has a mutation in ROS receptor. In some embodiments, the subject has a mutation in LTK receptor. In some embodiments, the subject has a mutation in ROR receptor. In some embodiments, the subject has a mutation in the MuSK receptor.

[0128] In some embodiments, the subject has a mutation in EGFR, provided that the subject also has a KRAS G12C mutation. In some embodiments, the subject has an EGFR mutation including an EGFR exon 19 deletion, an exon 20 insertion, an L858X mutation, a T790X mutation, a C797X mutation, a G719X mutation, a L861X mutation, an S768X mutation, an E709X mutation, or any combination thereof. In some embodiments, the subject has an EGFR mutation including an EGFR exon 19 deletion and / or an exon 20 insertion. In some embodiments, the subject has an EGFR exon 19 deletion. In some embodiments, the subject has an EGFR exon 20 insertion.

[0129] In some embodiments, the subject does not have a mutation in PTPN11, such as an E76K mutation.

[0130] In some embodiments, the subject has a solid tumor that has progressed or relapsed on or after at least one prior line of systemic therapy including a platinum-based doublet chemotherapy and / or an anti-PD-1 / PD-L1 therapy, each administered as a monotherapy or both administered in combination therapy.

[0131] In some embodiments, the subject has measurable disease according to Response Evaluation Criteria in Solid Tumors (RECIST). In some embodiments, treatment of the subject with a compound of Formula (I) or (10b) and sotolacib results in a measurable change in disease state according to RECIST.

[0132] In some embodiments, the subject has not previously participated in an interventional clinical study for a period of at least about four (4) weeks or five (5) half-lives of the agent used in the interventional clinical study, whichever is shorter, prior to initiation of treatment with the combination of a compound of Formula (I) or (10b) and sotolacib.

[0133] In some embodiments, the subject has not previously received radiation therapy or proton therapy prior to initiating treatment with the combination of a compound of Formula (I) or (10b) and sotolacib, including i) limited-scale radiation within a period of about one (1) week, or ii) radiation to more than about 30% of the bone marrow or extensive radiation within a period of about four (4) weeks.

[0134] In some embodiments, the subject has not taken or is not currently taking: a) one or more strong or moderate inducers or inhibitors of CYP3A4 and / or P-gp inducers (including herbal supplements or foods containing grapefruit juice, carambola, or citrus fruit) (e.g., as described in Example 3) within 14 days or five (5) half-lives, whichever is longer, prior to starting treatment with the combination of a compound of Formula (I) or (10b) and sotolacib; b) a drug that is a known substrate of CYP3A4 and / or P-gp within 14 days or five (5) half-lives, whichever is longer, prior to starting treatment with the combination of a compound of Formula (I) or (10b) and sotolacib; and / or one or more acid reducing agents, such as a proton pump inhibitor (PPI) or an H2 receptor antagonist, within 3 days of starting treatment with the combination of a compound of Formula (I) or (10b) and sotolacib.

[0135] In some embodiments, the subject does not have inadequate organ function, including adequate hematological function, renal function, hepatic function, and coagulation function, as defined below:

[0136] hematology

[0137] a. Absolute neutrophil count <1,500 / μL;

[0138] b. Platelets <100,000 / μL; and

[0139] c. Hemoglobin <9 g / dL without transfusion for ≤2 weeks or erythropoiesis-stimulating agents (eg, Epo, Procrit) for ≤6 weeks.

[0140] kidney

[0141] d. Serum creatinine >1.5×ULN unless creatinine clearance ≥40 mL / min (measured or calculated using the Cockcroft-Gault formula)

[0142] liver

[0143] e. Serum total bilirubin ≥2× institutional upper limit of normal (ULN) or ≥3.0× institutional ULN if the patient has a diagnosis of Gilbert syndrome or hemolytic anemia confirmed by the investigator; and

[0144] f. Aspartate aminotransferase / serum glutamate-oxaloacetate transaminase (AST / SGOT) and / or alanine aminotransferase / serum glutamate-pyruvate transaminase (ALT / SGPT)>2.5×ULN.

[0145] Blood clotting

[0146] g. International normalized ratio (INT) or prothrombin time (PT) >1.5×ULN, unless the patient is receiving anticoagulant therapy and as long as the PT or activated partial thromboplastin time (aPTT) is within the therapeutic range for the intended use of the anticoagulant; and

[0147] h. Activated partial thromboplastin time >1.5×ULN unless the patient is receiving anticoagulant therapy and as long as the PT or aPTT is within the therapeutic range for the intended use of the anticoagulant.

[0148] In some embodiments, the subject does not have active hepatitis B infection, hepatitis C infection, or human immunodeficiency virus (HIV) infection and a detectable viral load.

[0149] In some embodiments, the subject does not have a life-threatening disease, medical condition, active uncontrolled infection, or systemic organ dysfunction (eg, ascites, coagulopathy, or brain lesions).

[0150] In some embodiments, the subject does not have one or more heart-related diseases or is found to:

[0151] a) History of significant cardiovascular disease (e.g., cerebrovascular accident, myocardial infarction, or unstable angina) within the last 6 months before starting treatment;

[0152] b) Clinically significant heart disease, including New York Heart Association class II or higher heart failure;

[0153] c) History of left ventricular ejection fraction (LVEF) < 50% within 12 months before starting treatment;

[0154] d) Resting corrected QT interval (QTc) > 470 msec, averaged from three electrocardiograms (ECGs) using the provided ECG machine; and / or

[0155] e) Any clinically significant abnormality in the rhythm, conduction, or morphology of the resting ECG (e.g., third degree atrioventricular block, Mobitz type II heart block, ventricular arrhythmia, uncontrolled atrial fibrillation).

[0156] In some embodiments, the subject has not been diagnosed with another invasive malignancy within the past 3 years, with the proviso that the other invasive malignancy is not curatively treated non-melanoma skin cancer, superficial urothelial carcinoma, cervical carcinoma in situ, or any other curatively treated malignancy, and it is not expected that treatment for recurrence of the malignancy will not be required during the course of treatment with the combination of a compound of Formula (I) or (10b) and sotolacib.

[0157] In some embodiments, the subject does not have one or more untreated brain metastases from a non-brain tumor.

[0158] In some embodiments, subjects who have had brain metastases resected or have completed radiation therapy for at least 4 weeks prior to starting treatment with a compound of Formula (I) or (10b) and sotolacib combination therapy (e.g., Cycle 1, Day 1) are eligible, provided that the subject meets all of the following criteria prior to starting treatment: a) residual neurological symptoms related to CNS treatment ≤ Grade 2; b) taking a stable or decreasing dose of ≤ 10 mg prednisone (or equivalent) daily for at least 2 weeks prior to Cycle 1, Day 1, if applicable; and c) no new lesions on follow-up magnetic resonance imaging (MRI) within 4 weeks prior to Cycle 1, Day 1.

[0159] In some embodiments, the subject has not undergone major surgery within 4 weeks prior to enrollment in the combination treatment with a compound of Formula (I) or (10b) and sotolacib, with the proviso that the surgery or procedure was not a peripherally inserted central line placement, thoracentesis, abdominal puncture, tissue biopsy, or abscess drainage procedure.

[0160] In some embodiments, the subject has no history of allergy to sotolacilib or a compound of Formula (I) or (10b), an active or inactive excipient of sotolacilib or a compound of Formula (I) or (10b), or a drug of a similar chemical structure or class to sotolacilib or a compound of Formula (I) or (10b), depending on which combination is acceptable to the subject.

[0161] In some embodiments, the subject does not have one or more other activating mutations in BRAF V600X, PTPN11 (SHP2), and / or KRAS Q61X. In some embodiments, the subject does not have a tumor carrying one or more other activating mutations in BRAF V600X, PTPN11 (SHP2), and / or KRAS Q61X.

[0162] In some embodiments, the subject has not previously been treated with a KRAS G12C inhibitor (e.g., as described herein). In some embodiments, the subject has not previously been treated with sotolasib. In some embodiments, the subject has previously been treated with sotolasib. In some embodiments, the subject has previously been treated with a KRASG12C inhibitor other than sotolasib. In some embodiments, the subject has not previously been treated with a PTPN11 inhibitor (e.g., a SHP2 inhibitor), provided that the PTPN11 inhibitor is not a compound of formula (I) or (10b). In some embodiments, the subject has not previously been treated with a PTPN11 inhibitor selected from the following: TNO-155, RMC-4630, RLY-1971, JAB-3068, JAB-3312, PF-07284892, and ERAS601. In some embodiments, the subject has not previously been treated with a compound of formula (I) or (10b). In some embodiments, the subject has been previously treated with a SHP2 inhibitor, including any one of TNO-155, RMC-4630, RLY-1971, JAB-3068, JAB-3312, PF-07284892, ERAS601, and a compound of formula (I) or (10b). In some embodiments, the subject has been previously treated with a compound of formula (I) or (10b). In some embodiments, the subject has been previously treated with a PTPN11 inhibitor other than a compound of formula (I) or (10b).

[0163] In some embodiments, the subject does not have a gastrointestinal disorder that may prevent absorption of the compound of Formula (I) or (10b) (eg, post-gastrectomy, broken bowel syndrome, uncontrolled Crohn's disease, celiac disease with villous atrophy, or chronic gastritis).

[0164] In some embodiments, the subject is not on dialysis.

[0165] In some embodiments, the subject has no history of allogeneic bone marrow transplantation.

[0166] Further inclusion and exclusion criteria for subjects who may benefit from treatment with a compound of Formula (I) or (10b) and sotolacib in combination are described in Example 3, such as subjects enrolled in a Phase 1 study of a combination of the SHP2 inhibitor compound (10b) and sotolacib.

[0167] In some embodiments, the subject meets all inclusion criteria of 1) to 11) as described in Example 3. In some embodiments, the subject meets all inclusion criteria of 1) to 11) as described in Example 3, with the limitation that the subject does not meet any of the exclusion criteria of 1) to 17) as described in Example 3.

[0168] III-4: Treatment cycle and dose adjustment

[0169] The treatment of the combination of Formula (I) or (10b) compound and Sotolacib can include one or more treatment cycles (e.g., at least 1, 2, 3 or more treatment cycles). In some embodiments, the treatment includes one or more treatment cycles (e.g., at least 1, 2, 3 or more treatment cycles). In some embodiments, the treatment includes at least 2, 3 or more treatment cycles. In some embodiments, the treatment includes 2 to 3 treatment cycles. In some embodiments, the treatment includes 3 treatment cycles. In some embodiments, the treatment includes more than 3 treatment cycles.

[0170] In some embodiments, each of the one or more treatment cycles has a duration of about 28 days; and the compound of formula (I) or (10b) is administered daily. In some embodiments, each of the one or more treatment cycles has a duration of about 28 days; and sotolacib is administered daily. In some embodiments, each of the one or more treatment cycles has a duration of about 28 days; the compound of formula (I) or (10b) is administered daily; and sotolacib is administered daily.

[0171] Treatment may include a dose escalation period during which the dose of the compound of Formula (I) or (10b) or sotolacib may be adjusted (e.g., dose increased or decreased) or maintained following the previous treatment cycle. Dose adjustments may be based at least in part on safety evaluations (e.g., dose-limiting toxicity (DLT) assessments).

[0172] In some embodiments, the subject is initially treated with a compound of Formula (I) or (10b) and sotolacib at a first compound dose level and a first sotolacib dose level, and is subsequently treated with a second compound dose level and a second sotolacib dose level, wherein the second compound dose level is different from the first compound dose level and / or the second sotolacib dose level is different from the first sotolacib dose level. In some embodiments, the second sotolacib dose level is lower than the first sotolacib dose level. In some embodiments, the second sotolacib dose level is higher than the first sotolacib dose level. In some embodiments, the second compound dose level is lower than the first compound dose level. In some embodiments, the second compound dose level is higher than the first compound dose level.

[0173] In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose escalations (e.g., dose increases), dose maintenance, or dose reductions (e.g., dose reductions) of sotolacibu and / or a compound of formula (I) or (10b). In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose escalations, dose maintenance, or dose reductions of sotolacibu. In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose reductions of sotolacibu. In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose escalations of sotolacibu. In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose escalations, dose maintenance, or dose reductions of a compound of formula (I) or (10b). In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose decrements (e.g., dose reductions) of the compound of formula (I) or (10b). In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I.) or (10b) comprises one or more dose escalations (e.g., dose increases) of the compound of formula (I) or (10b). In some embodiments, the administration of the combination of sotolacibu and a compound of formula (I) or (10b) comprises one or more dose escalations (e.g., dose increases), dose maintenance, or dose decrements (e.g., dose reductions) of sotolacibu and / or a compound of formula (I) or (10b), each of which is determined by a safety or dose-limiting toxicity (DLT) assessment (e.g., associated with a subject cohort). In some embodiments, administration of the combination of sotolacib and a compound of Formula (I) or (10b) comprises one or more dose escalations, dose maintenance, or dose reductions of the compound of Formula (I) or (10b), each of which is determined by a dose-limiting toxicity (DLT) assessment, as described in Examples 3 and Figure 6 Described in .

[0174] In some embodiments, when the dose-limiting toxicity (DLT) rate is less than, for example, about 19.7%, as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises dose escalation after the previous treatment cycle. In some embodiments, when the dose-limiting toxicity (DLT) rate is less than, for example, about 19.7%, as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises dose escalation in a second treatment cycle after a first treatment cycle. In some embodiments, when the dose-limiting toxicity (DLT) rate is less than, for example, about 19.7%, as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises dose escalation in a third treatment cycle after a second treatment cycle.

[0175] In some embodiments, when the dose-limiting toxicity (DLT) rate is greater than, for example, about 29.8%, as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises a dose reduction after the previous treatment cycle. In some embodiments, when the dose-limiting toxicity (DLT) rate is greater than, for example, about 29.8%, as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises a dose reduction in a second treatment cycle after a first treatment cycle. In some embodiments, when the dose-limiting toxicity (DLT) rate is greater than, for example, about 29.8%, as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises a dose reduction in a third treatment cycle after a second treatment cycle.

[0176] In some embodiments, when the dose-limiting toxicity (DLT) rate is in the range of about 21.9% to about 29.8% as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises a dose maintenance after the previous treatment cycle. In some embodiments, when the dose-limiting toxicity (DLT) rate is in the range of about 21.9% to about 29.8% as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises a dose maintenance in the second treatment cycle after the first treatment cycle. In some embodiments, when the dose-limiting toxicity (DLT) rate is in the range of about 21.9% to about 29.8% as determined by a DLT assessment (e.g., associated with a subject cohort), the administration of a compound of Formula (I) or (10b) comprises a dose maintenance in the third treatment cycle after the second treatment cycle.

[0177] After the dose escalation phase, treatment also includes a dose expansion / optimization phase. In some embodiments of the dose expansion / optimization phase, the compound of formula (I) or (10b) is administered at a dose regimen determined in the dose escalation phase (eg, dose regimen 1 or dose regimen 2).

[0178] In some embodiments, the administration of a compound of formula (I) or (10b) includes one or more dose adjustments. In some embodiments, the administration of a compound of formula (I) or (10b) includes one or more dose adjustments during a dose expansion / optimization phase. In some embodiments, the administration of a compound of formula (I) or (10b) optionally includes one or more dose adjustments during a dose expansion / optimization phase; and the one or more dose adjustments are determined based on a safety review committee (SRC) safety assessment.

[0179] In some embodiments, dose adjustments, delays, and interruptions of the compound of Formula (I) or (10b) and / or sotolacib are further based on the criteria of Example 3.

[0180] III-5: Therapeutically effective amount / administration

[0181] The compound of formula (I) or (10b) and Sotolacib can be provided in a combined therapeutically effective amount or a synergistically effective amount, or each of them can be used in a dose different from that when each is used alone. In some embodiments, the compound of formula (I) or (10b) and Sotolacib are provided in a combined therapeutically effective amount. In some embodiments, the compound of formula (I) or (10b) and Sotolacib are provided in a synergistically effective amount.

[0182] In some embodiments, the compound of formula (I) or (10b) and / or sotolacib are used at a dosage different from that when used alone (e.g., in a monotherapy treatment form). In some embodiments, the compound of formula (I) or (10b) and sotolacib are each used at a dosage different from that when each is used alone. In some embodiments, the compound of formula (I) or (10b) and sotolacib are each used at a dosage lower than that when each is used alone. In some embodiments, the compound of formula (I) or (10b) is used at a dosage lower than that when used alone. In some embodiments, sotolacib is used at a dosage lower than that when used alone. In some embodiments, the compound of formula (I) or (10b) is used at a dosage higher than that when used alone. In some embodiments, sotolacib is used at a dosage higher than that when used alone.

[0183] The compound of formula (I) or (10b) and Sotolacib can be administered concomitantly or sequentially. In some embodiments, the compound of formula (I) or (10b) and Sotolacib are administered concomitantly. In some embodiments, the compound of formula (I) or (10b) and Sotolacib are administered in a pharmaceutical composition, which comprises the compound of formula (I) or (10b) and Sotolacib. In some embodiments, the compound of formula (I) or (10b) and Sotolacib are administered sequentially. In some embodiments, the compound of formula (I) or (10b) is administered before Sotolacib is administered. In some embodiments, the compound of formula (I) or (10b) is administered after Sotolacib is administered.

[0184] In some embodiments, the therapeutically effective amount of a compound of formula (I) or (10b) is a total daily dose of no more than about 2000 mg, on a salt-free and water-free basis. In some embodiments, the therapeutically effective amount of the compound of formula (I) or (10b) is from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 450 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 550 mg to about 1000 mg, from about 600 mg to about 1000 mg, from about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 100 mg to about 700 mg, from about 150 mg to about 700 mg, from about 200 mg to about 700 mg, from about 250 mg to about 700 mg, from about 300 mg to about 1000 mg. From about 3 mg to about 700 mg, from about 400 mg to about 700 mg, from about 450 mg to about 700 mg, from about 500 mg to about 700 mg, from about 550 mg to about 700 mg, from about 100 mg to about 550 mg, from about 150 mg to about 550 mg, from about 200 mg to about 550 mg, from about 250 mg to about 550 mg, from about 300 mg to about 550 mg, from about 350 mg to about 550 mg, from about 400 mg to about 550 mg, from about 450 mg to about 550 mg, from about 100 mg to about 400 mg, from about 150 mg to about 400 mg, from about 200 mg to about 400 mg, from about 250 mg to about 400 mg, or from about 300 mg to about 400 mg, on a salt-free and water-free basis, or any effective range therein.

[0185] In some embodiments, the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg, on a salt-free and water-free basis, or any effective range therein.

[0186] In some embodiments, the therapeutically effective amount of the compound of formula (10b) is about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, or about 1000 mg of the total daily dose of the compound of formula (10b), calculated on a salt-free and anhydrous basis. In some embodiments, the therapeutically effective amount of the compound of formula (10b) is about 250 mg, about 400 mg, or about 550 mg of the total daily dose of the compound of formula (10b), calculated on a salt-free and anhydrous basis. In some embodiments, the therapeutically effective amount is about 250 mg of the total daily dose of the compound of formula (10b), calculated on a salt-free and anhydrous basis. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated on a salt-free and water-free basis. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated on a salt-free and water-free basis.

[0187] In some embodiments, the therapeutically effective amount of sotolacib is from about 120 mg to about 960 mg, from about 240 mg to about 960 mg, from about 360 mg to about 960 mg, from about 480 mg to about 960 mg, from about 600 mg to about 960 mg, from about 720 mg to about 960 mg, from about 840 mg to about 960 mg, from about 120 mg to about 840 mg, from about 240 mg to about 840 mg, from about 360 mg to about 840 mg, from about 480 mg to about 840 mg, from about 600 mg to about 840 mg, from about 720 mg to about 960 mg, from about 840 mg to about 960 mg. In some embodiments, the total daily dose of sotolacib can be from about 20 mg, from about 240 mg to about 720 mg, from about 360 mg to about 720 mg, from about 480 mg to about 720 mg, from about 600 mg to about 720 mg, from about 120 mg to about 600 mg, from about 240 mg to about 600 mg, from about 360 mg to about 600 mg, from about 480 mg to about 600 mg, from about 120 mg to about 480 mg, from about 240 mg to about 480 mg, from about 360 mg to about 480 mg, from about 120 mg to about 360 mg, from about 240 mg to about 360 mg, or from about 120 mg to about 240 mg.

[0188] In some embodiments, the therapeutically effective amount of sotolaciib is a total daily dose of about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg of sotolaciib.

[0189] In some embodiments, the therapeutically effective amount of the compound of formula (10b) is a total daily dose of no more than about 2000 mg, calculated on a salt-free and anhydrous basis; and the therapeutically effective amount of sotolacib is a total daily dose of no more than about 960 mg. In some embodiments, the therapeutically effective amount of the compound of formula (10b) is a total daily dose of no more than about 2000 mg, calculated on a salt-free and anhydrous basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 960 mg. In some embodiments, the therapeutically effective amount of the compound of formula (10b) is from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 450 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 550 mg to about 1000 mg, from about 600 mg to about 1000 mg, from about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 100 mg to about 700 mg, from about 150 mg to about 700 mg, from about 200 mg to about 700 mg, from about 250 mg to about 700 mg, from about 300 mg to about 700 mg, from about from about 350 mg to about 700 mg, from about 400 mg to about 700 mg, from about 450 mg to about 700 mg, from about 500 mg to about 700 mg, from about 550 mg to about 700 mg, from about 100 mg to about 550 mg, from about 150 mg to about 550 mg, from about 200 mg to about 550 mg, from about 250 mg to about 550 mg, from about 300 mg to about 550 mg, from about 350 mg to about 550 mg, from about 400 mg to about 550 mg, from about 450 mg to about 550 mg, from about 100 mg to about 400 mg, from about 150 mg to about 400 mg, from about 200 mg to about 400 mg, from about 250 mg to about 400 mg, or from about 300 mg to about 400 mg of the compound of formula (10b), on a salt-free and water-free basis, or any effective range therebetween;and a therapeutically effective amount of sotolacib is from about 120 mg to about 960 mg, from about 240 mg to about 960 mg, from about 360 mg to about 960 mg, from about 480 mg to about 960 mg, from about 600 mg to about 960 mg, from about 720 mg to about 960 mg, from about 840 mg to about 960 mg, from about 120 mg to about 840 mg, from about 240 mg to about 840 mg, from about 360 mg to about 840 mg, from about 480 mg to about 840 mg, from about 600 mg to about 840 mg, from about 720 mg to about 840 mg, from about 120 mg to about 720 mg , a total daily dose of sotolacib from about 240 mg to about 720 mg, from about 360 mg to about 720 mg, from about 480 mg to about 720 mg, from about 600 mg to about 720 mg, from about 120 mg to about 600 mg, from about 240 mg to about 600 mg, from about 360 mg to about 600 mg, from about 480 mg to about 600 mg, from about 120 mg to about 480 mg, from about 240 mg to about 480 mg, from about 360 mg to about 480 mg, from about 120 mg to about 360 mg, from about 240 mg to about 360 mg, or from about 120 mg to about 240 mg. In some embodiments, the therapeutically effective amount is a total daily dose of the compound of formula (10b) of about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 250 mg to about 700 mg, from about 250 mg to about 550 mg, from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg, on a salt-free and water-free basis, or any effective range therebetween; and the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg. In some embodiments, the therapeutically effective amount is a total daily dose of the compound of formula (10b) from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg, on a salt-free and water-free basis, or any effective range therebetween; and the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg. ;

[0190] In some embodiments, the therapeutically effective amount of the compound of formula (10b) is a total daily dose of about 250 mg, about 400 mg, or about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 960 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 960 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated on a salt-free and anhydrous basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 960 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated on a salt-free and anhydrous basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 840 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated on a salt-free and anhydrous basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 840 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated on a salt-free and anhydrous basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 840 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 720 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 720 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 720 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 600 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), on a salt-free and water-free basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 600 mg.In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 600 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 480 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 480 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 480 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 360 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 360 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 360 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 240 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 240 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 240 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg. In some embodiments, the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (10b), on a salt-free and water-free basis; and the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg.

[0191] In general, the compound of formula (I) or (10b) can be administered orally. In some embodiments, the compound of formula (10b) is administered orally. In some embodiments, the compound of formula (10b) is administered orally in a tablet formulation.

[0192] In general, sotolacib can be administered orally. In some embodiments, sotolacib is administered orally. In some embodiments, sotolacib in a tablet formulation is administered orally.

[0193] In general, the compound of formula (I) or (10b) can be administered once or more (e.g., 2, 3, 4 or more times) daily. In some embodiments, the compound of formula (10b) is administered once, twice, three times or four times daily. In some embodiments, the compound of formula (10b) is administered once daily. In some embodiments, the compound of formula (10b) is administered twice daily. In some embodiments, the compound of formula (10b) is administered every other day. In some embodiments, the compound of formula (10b) is administered as follows: four days of administration and three days of withdrawal (e.g., the compound is administered for four consecutive days and then not administered for three consecutive days), five days of administration and two days of withdrawal, two days of administration and five days of withdrawal, one week of administration and one week of withdrawal, two weeks of administration and one week of withdrawal, three weeks of administration and one week of withdrawal, or a similar schedule.

[0194] In general, sotolacib can be administered once, twice, or more times (e.g., 2, 3, 4 or more times) daily. In some embodiments, sotolacib is administered once daily.

[0195] In some embodiments, the compound of formula (I) or (10b) and sotolacib are each administered orally. In some embodiments, the compound of formula (10b) and sotolacib are each administered orally. In some embodiments, the compound of formula (I) or (10b) is administered once daily; and sotolacib is administered once daily. In some embodiments, the compound of formula (10b) is administered once daily; and sotolacib is administered once daily.

[0196] The compound of formula (I) or (10b) may be in an oral dosage form of one or more dosage strengths, wherein the compound of formula (I) or (10b) is present in an amount of at least about 5 mg, 10 mg, 20 mg, 30 mg, 50 mg, 100 mg, 120 mg, 180 mg, 200 mg, 300 mg, 400 mg or 500 mg, based on salt-free and water-free. In some embodiments, the oral dosage form is a tablet formulation of one or more dosage strengths. In some embodiments of the tablet formulation, the compound of formula (10b) is present in each tablet in an amount of from 30 to 1000 mg, from 30 to 750 mg, from 30 to 500 mg, from 30 to 200 mg, from 30 to 180 mg, from 30 to 120 mg, from 30 to 90 mg, from 50 to 1000 mg, from 50 to 750 mg, from 50 to 500 mg, from 50 to 250 mg, from 100 to 1000 mg, from 100 to 750 mg, from 1 From 100 to 250 mg, from 200 to 1000 mg, from 200 to 750 mg, from 200 to 500 mg, from 300 to 1000 mg, from 300 to 750 mg, from 300 to 500 mg, from 400 to 1000 mg, from 400 to 750 mg, from 500 to 1000 mg, from 500 to 750 mg, from 600 to 1000 mg, from 5 to 250 mg or from 5 to 100 mg, calculated without salt and water.

[0197] In some embodiments of the tablet formulation, the compound of formula (10b) is present in each tablet in an amount of about 5 mg, 10 mg, 30 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg or 1000 mg, in a salt-free and anhydrous amount. In some embodiments of the tablet formulation, the compound of formula (10b) is present in each tablet in an amount of about 30 mg, 50 mg or 100 mg, in a salt-free and anhydrous amount. In some embodiments of the tablet formulation, the compound of formula (10b) is present in each tablet in an amount of about 30 mg, in a salt-free and anhydrous amount. In some embodiments of the tablet formulation, the compound of formula (10b) is present in each tablet in an amount of about 50 mg, in a salt-free and anhydrous amount. In some embodiments of tablet formulations, the compound of formula (10b) is present in each tablet in an amount of about 100 mg on a salt-free and water-free basis.

[0198] In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of no more than about 2000 mg of the compound of formula (10b). In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 450 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 550 mg to about 1000 mg, from about 600 mg to about 1000 mg, from about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 100 mg to about 700 mg, from about 150 mg to about 700 mg, from about 200 mg to about 700 mg, from about 250 mg to about 700 mg, from about 1 From about 300 mg to about 700 mg, from about 350 mg to about 700 mg, from about 400 mg to about 700 mg, from about 450 mg to about 700 mg, from about 500 mg to about 700 mg, from about 550 mg to about 700 mg, from about 100 mg to about 550 mg, from about 150 mg to about 550 mg, from about 200 mg to about 550 mg, from about 250 mg to about 550 mg, from about 300 mg to about 550 mg, from about 350 mg to about 550 mg, from about 400 mg to about 550 mg, from about 450 mg to about 550 mg, from about 100 mg to about 400 mg, from about 150 mg to about 400 mg, from about 200 mg to about 400 mg, from about 250 mg to about 400 mg, or from about 300 mg to about 400 mg of the compound of formula (10b), calculated on a salt-free and water-free basis. In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 250 mg to about 700 mg, from about 250 mg to about 550 mg, from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg of the compound of formula (10b), calculated on a salt-free and anhydrous basis. In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg of the compound of formula (10b), calculated on a salt-free and anhydrous basis.In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of about 250 mg, about 400 mg, about 550 mg of the compound of formula (10b) on a salt-free and water-free basis.

[0199] Sotolacib can be an oral dosage form of one or more dosage strengths. In some embodiments, the oral dosage form of Sotolacib is a tablet formulation of one or more dosage strengths. In some embodiments, Sotolacib is provided in a tablet formulation containing about 120 mg of Sotolacib in each tablet. In some embodiments, Sotolacib is provided in a tablet formulation containing about 60 mg, 120 mg, 240 mg, 360 mg or more of Sotolacib in each tablet.

[0200] In some embodiments, sotolacib is administered once daily to provide a total daily dose of no more than about 2000 mg of sotolacib. In some embodiments, sotolacib is administered once daily to provide a total daily dose of no more than about 960 mg. In some embodiments, sotolacib is administered once daily to provide a total daily dose of from about 120 mg to about 960 mg, such as about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg.

[0201] In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of no more than about 2000 mg of the compound of formula (10b); and sotolacib is administered once daily to provide a total daily dose of no more than about 960 mg. In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 250 mg to about 700 mg, from about 250 mg to about 550 mg, from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg of the compound of formula (10b), on a salt-free and water-free basis; and sotolacib is administered once daily to provide a total daily dose of from about 120 mg to about 960 mg, such as about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg. In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and sotolacib is administered once daily to provide a total daily dose of from about 120 mg to about 960 mg, such as about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg. In some embodiments, the compound of formula (10b) is administered once daily to provide a total daily dose of about 250 mg, about 400 mg, about 550 mg of the compound of formula (10b), calculated as salt-free and anhydrous; and sotolacib is administered once daily to provide a total daily dose of about 960 mg.

[0202] In some embodiments, during each of one or more treatment cycles, the compound of formula (10b) is administered once daily, as described herein. In some embodiments, during each of one or more treatment cycles, sotolacib is administered once daily, as described herein. In some embodiments, during each of one or more treatment cycles, the compound of formula (10b) and sotolacib are administered once daily, as described herein.

[0203] In general, it is recommended that the compound of formula (10b) be administered to a subject without food (e.g., after an overnight fast (minimum 8 hours), followed by a 2-hour fast after taking the dose). Subjects may drink water except for one (1) hour before and after administration, and water (e.g., 240 mL) is given to the subject at the time of administration. In some embodiments, the compound of formula (10b) is administered to a subject without food for at least about 8 hours before administration and at least about 2 hours after administration.

[0204] In some embodiments, sotolacib is administered once daily within about 5 minutes after administration of the compound of formula (10b).

[0205] III-6: Effectiveness

[0206] "A Phase 1 study of the combination of the SHP2 inhibitor compound of formula (10b.) and the KRAS G12C inhibitor sotolacib in patients with advanced or metastatic non-small cell lung cancer harboring a KRAS G12C mutation" can evaluate the safety, tolerability and efficacy of the combination of the compound of formula (10b) and sotolacib in reducing or stabilizing solid tumors in subjects, as outlined in Example 3.

[0207] In various embodiments, the subject is administered therapy for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 15 months, at least 18 months, at least 21 months, or at least 23 months, e.g., for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 15 months, 18 months, 21 months, or 24 months. In various embodiments, the subject is administered therapy for at least 1 month. In various embodiments, the subject is administered therapy for at least 3 months. In various embodiments, the subject is administered therapy for at least 6 months. In various embodiments, the subject is administered therapy for at least 8 months.

[0208] A subject may respond to therapy as measured by at least one stable disease (SD), as determined by the Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 protocol (Eisenhauer et al., Eur J Cancer; 2009; 45(2): 228-247). RECIST v1.1 will be discussed in detail in the following examples. At least stable disease refers to stable disease that has shown a partial response (PR) or has shown a complete response (CR) (i.e., "at least SD" = SD + PR + CR, generally referred to as disease control). In various embodiments, stable disease is neither sufficiently reduced to qualify as a partial response (PR) nor sufficiently increased to qualify as progressive disease (PD). In various embodiments, a subject shows at least a partial response (i.e., "at least pR" = PR + CR, generally referred to as an objective response).

[0209] Response can be measured by one or more of: reduction in tumor size, inhibition or reduction in tumor growth, reduction in target or tumor lesions, delay in time to progression, absence of new tumors or lesions, reduction in new tumor formation, prolonged survival or progression-free survival (PFS), and absence of metastasis. In various embodiments, the progression of the subject's disease can be assessed by measuring tumor size, tumor lesions, or the formation of new tumors or lesions, by assessing the subject using computed tomography (CT) scans, positron emission tomography (PET) scans, magnetic resonance imaging (MRI) scans, X-rays, ultrasound, or some combination thereof.

[0210] The progression-free survival (PFS) can be assessed according to the description in the RECIST 1.1 scheme. In various embodiments, the subject shows a PFS of at least 1 month. In various embodiments, the subject shows a PFS of at least 3 months. In some embodiments, the subject shows a PFS of at least 6 months.

[0211] The combined administration of a therapeutically effective amount of a compound of formula (I) or (10b) and a therapeutically effective amount of sotolacib can reduce or substantially eliminate cancer or solid tumors in a subject. In some embodiments, the combination of a therapeutically effective amount of formula (I) or (10b) and sotolacib substantially eliminates solid tumors. In some embodiments, the combination of a therapeutically effective amount of formula (I) or (10b) and sotolacib reduces the volume of a solid tumor by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or more. In some embodiments, a therapeutically effective amount of the combination of Formula (I) or (10b) and sotolacib reduces the volume of a solid tumor by from about 10% to about 90%, from about 10% to about 80%, from about 10% to about 70%, from about 10% to about 60%, from about 10% to about 50%, from about 10% to about 40%, from about 10% to about 30%, from about 10% to about 20%, from about 20% to about 90%, from about 20% to about 80%, from about 20% to about 70%, from about 20% to about 60%, from about 20% to about 50%, from about 20% to about 40%, from about 20% to about 30%, from about 10% to about 20%, from about 20% to about 90%, from about 20% to about 80%, from about 20% to about 70%, from about 20% to about 60%, from about 20% to about 50%, from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 90%, from about 30% to about From about 80%, from about 30% to about 70%, from about 30% to about 60%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 90%, from about 40% to about 80%, from about 40% to about 70%, from about 40% to about 60%, from about 40% to about 50%, from about 50% to about 90%, from about 50% to about 80%, from about 50% to about 70%, from about 50% to about 60%, from about 60% to about 90%, from about 60% to about 80%, from about 60% to about 70%, from about 70% to about 90%, from about 70% to about 80%, from about 80% to about 90%, or any range therein. In some embodiments, the combination of a therapeutically effective amount of Formula (I) or (10b) and sotolacib reduces the volume of a solid tumor by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%.

[0212] The combination of a therapeutically effective amount of a compound of formula (I) or (10b) and a therapeutically effective amount of sotolacib can stabilize a cancer or solid tumor in a subject. In some embodiments, the combination of a therapeutically effective amount of a compound of formula (I) or (10b) and sotolacib stabilizes a solid tumor.

[0213] The combination of a therapeutically effective amount of a compound of formula (I) or (10b) and a therapeutically effective amount of Sotolacib can maintain the reduction or stabilization of cancer or solid tumors over a period of time (e.g., 1 to 12 months). In some embodiments, a combination of a therapeutically effective amount of a compound of formula (I) or (10b) and Sotolacib is used, and the solid tumor is reduced or stabilized for at least about one month. In some embodiments, a combination of a therapeutically effective amount of a compound of formula (10b) and Sotolacib is used, and the solid tumor is reduced or stabilized for at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months. In some embodiments, the solid tumor is reduced or stabilized from about 1 to about 12 months, from about 1 to about 6 months, from about 1 to about 3 months, or from about 1 to about 2 months.

[0214] In some embodiments, the subject is further evaluated by one or more tests (e.g., as described herein) to provide an overall assessment including plasma pharmacokinetic and / or pharmacodynamic properties. Examples of such tests are described, e.g., in Example 3.

[0215] In some embodiments, the subject's one or more biomarkers are further evaluated to determine the correlation of the one or more biomarkers with anti-tumor response. Examples of such evaluations are described in Example 3.

[0216] IV. Composition

[0217] Compositions of compounds of formula (I) or (10b)

[0218] The oral dosage form comprising the compound of formula (I) or (10b) can be any oral dosage form, which comprises one or more pharmaceutically acceptable carriers and / or excipients. Oral preparations include tablets, pills, powders, dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc., which are suitable for patient ingestion.

[0219] For preparing oral dosage forms comprising a compound of formula (I) or (10b), a pharmaceutically acceptable carrier can be a solid or a liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances which may also serve as a diluent, flavoring agent, binder, preservative, tablet disintegrating agent, or encapsulating material. Details of formulation and administration techniques are described in detail in the scientific and patent literature, see, for example, the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

[0220] In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and prepared in the shape and size desired.

[0221] Powders, capsules and tablets preferably contain from 5% to 70% of the active pharmaceutical ingredient (e.g., a compound of formula (I) or (10b)), or about 10% to about 70% of the active pharmaceutical ingredient. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compound and an encapsulating material as a carrier, providing a capsule in which the active ingredient is surrounded by a carrier with or without other excipients, thereby combining with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets and lozenges can be used as solid dosage forms suitable for oral administration.

[0222] Suitable solid excipients include, but are not limited to, magnesium carbonate; magnesium stearate; talc; pectin; dextrin; starch; tragacanth; low melting wax; cocoa butter; carbohydrates; sugars, including but not limited to lactose, sucrose, mannitol or sorbitol, starch from corn, wheat, rice, potato or other plants; celluloses, such as methylcellulose, hydroxypropylmethylcellulose or sodium carboxymethylcellulose; and gums, including acacia and tragacanth; and proteins, including but not limited to gelatin and collagen. If desired, disintegrating agents or fluxing agents may be added, such as cross-linked polyvinyl pyrrolidone, agar, alginic acid or its salt, such as sodium alginate.

[0223] The dragee core has a suitable coating, such as a concentrated sugar solution, which may also contain gum arabic, talc, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol and / or titanium dioxide, lacquer solution and a suitable organic solvent or solvent mixture. Dyes or pigments may be added to the tablet or dragee coating for product identification or to characterize the amount (i.e., dosage) of the active compound. The pharmaceutical preparation in dosage form may also be used orally, for example, using push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and coatings such as glycerol or sorbitol. Push-fit capsules may contain active pharmaceutical ingredients mixed with fillers or binders (such as lactose or starch), lubricants (such as talc or magnesium stearate) and optionally selected stabilizers. In soft capsules, the active pharmaceutical ingredient may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin or liquid polyethylene glycol, with or without a stabilizer.

[0224] To prepare suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted and the active drug ingredient is dispersed uniformly therein by stirring. The molten homogeneous mixture is then poured into conveniently sized molds, allowed to cool, and thereby to solidify.

[0225] Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water / propylene glycol solutions.

[0226] Aqueous solutions suitable for oral use can be prepared by dissolving the active pharmaceutical ingredient in water and adding suitable colorants, flavoring agents, stabilizers and thickeners as needed. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely powdered active ingredient with the following in water: viscous materials, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth gum and gum arabic, and dispersants or wetting agents, such as naturally occurring phospholipids (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide with long-chain aliphatic alcohols (e.g., 17-ethyleneoxy cetyl alcohol), condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol (e.g., polyoxyethylene sorbitol monooleate) or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives (such as ethyl or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents and one or more sweetening agents (such as sucrose, aspartame or saccharin). The formulation may be adjusted according to the osmotic pressure.

[0227] Also included are solid form preparations that are intended to be converted into liquid form preparations for oral administration shortly before use. Such liquid forms include solutions, suspensions, and emulsions. In addition to the active ingredient, these preparations may also include coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, cosolvents, etc.

[0228] Oily suspensions can be prepared by suspending the active pharmaceutical ingredient in a vegetable oil (such as peanut oil, olive oil, sesame oil or coconut oil) or a mineral oil (such as liquid paraffin) or a mixture of these oils. Oily suspensions may contain thickeners, such as beeswax, hard paraffin or cetyl alcohol. Sweeteners may be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose. These preparations may be preserved by the addition of antioxidants (such as ascorbic acid). As an example of an injectable oily carrier, see Minto, J. Pharmacol. Exp. Ther. 281: 93-102, 1997. Pharmaceutical preparations containing compounds of formula (I) or (10b) may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil or a mineral oil, as described above, or a mixture of these. Suitable emulsifiers include naturally occurring gums such as gum arabic and gum tragacanth, naturally occurring phospholipids 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, such as in the formulation of syrups and elixirs. Such formulations may also contain analgesics, preservatives or coloring agents.

[0229] The compositions of the present disclosure (including compounds of formula (I) or (10b)) can be prepared into a variety of oral, parenteral and topical dosage forms. Oral preparations include tablets, pills, powders, dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc., suitable for ingestion by patients. The compositions of the present disclosure can also be administered by injection, i.e., intravenously, intramuscularly, intradermally, subcutaneously, intraduodenally or intraperitoneally. In addition, the compositions described herein can be administered by inhalation, such as intranasally. In addition, the compositions of the present disclosure can be administered transdermally. The compositions of the present disclosure can also be administered by intraocular, intravaginal and intrarectal routes, including suppositories, insufflations, powders and aerosol formulations (e.g., steroid inhalers, see Rohatagi, J. Clin. Pharmacol. 35: 1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75: 107-111, 1995).

[0230] For the preparation of pharmaceutical compositions of the present disclosure (including compounds of Formula (I) or (10b)), pharmaceutically acceptable carriers may be solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier may be one or more substances which may also serve as a diluent, flavoring agent, binder, preservative, tablet disintegrant, or encapsulating material. Details of formulation and administration techniques are described in detail in the scientific and patent literature, see, for example, the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

[0231] In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size specified.

[0232] Powders, capsules and tablets preferably contain from about 5% to about 70% active compound, such as from about 10% to about 70% active compound (e.g., compound of formula (I) or (10b)). Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compound with encapsulating material as a carrier, providing a capsule in which the active ingredient, with or without other excipients, is surrounded by a carrier, thereby being combined with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets and lozenges can be used as solid dosage forms suitable for oral administration.

[0233] Suitable solid excipients include, but are not limited to, magnesium carbonate; magnesium stearate; talc; pectin; dextrin; starch; tragacanth; low melting wax; cocoa butter; carbohydrates; sugars, including but not limited to lactose, sucrose, mannitol or sorbitol, starch from corn, wheat, rice, potato or other plants; celluloses, such as methylcellulose, hydroxypropylmethylcellulose or sodium carboxymethylcellulose; and gums, including acacia and tragacanth; and proteins, including but not limited to gelatin and collagen. Disintegrators or solubilizers may be added, such as cross-linked polyvinyl pyrrolidone, agar, alginic acid or its salts, such as sodium alginate.

[0234] The dragee core has a suitable coating, such as a concentrated sugar solution, which may also contain gum arabic, talc, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol and / or titanium dioxide, lacquer solution and a suitable organic solvent or solvent mixture. Dyes or pigments may be added to the tablet or dragee coating for product identification or characterization of the amount (i.e., dosage) of the active compound. The pharmaceutical preparation of the dosage form may also be used orally, such as push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and coatings such as glycerol or sorbitol. The push-fit capsule may contain an active compound (e.g., a compound of formula (I) or (10b) and sotolacib) mixed with a filler or adhesive (e.g., lactose or starch), a lubricant (e.g., talc or magnesium stearate) and optionally a selected stabilizer. In soft capsules, the active compounds (eg, compounds of formula (I) or (10b) and sotolacib) can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycol, with or without stabilizers.

[0235] To prepare suppositories, a low melting point wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active compound (e.g., a compound of formula (I) or (10b)) is dispersed uniformly therein by stirring. The molten homogeneous mixture is then poured into conveniently sized molds, allowed to cool, and thereby solidify.

[0236] Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water / propylene glycol solutions. For parenteral injection, liquid preparations can be formulated as solutions in aqueous polyethylene glycol solution.

[0237] Aqueous solutions suitable for oral use can be prepared by dissolving the active compound (eg, a compound of formula (I) or (10b)) as defined and described herein in water and adding, optionally, suitable colorants, flavoring agents, stabilizers and thickening agents. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely powdered active component in water with a viscous material such as a natural or synthetic gum, resin, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinyl pyrrolidone, gum tragacanth and gum acacia, and a dispersing or wetting agent such as a naturally occurring phosphatide (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide with long chain aliphatic alcohols (e.g., heptadecaethyleneoxycetyl alcohol), condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol (e.g., polyoxyethylene sorbitan monooleate), or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol anhydride (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives (such as ethyl or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents and one or more sweetening agents (such as sucrose, aspartame or saccharin). The formulation may be adjusted according to the osmotic pressure.

[0238] Also included are solid form preparations that are intended to be converted into liquid form preparations for oral administration shortly before use. Such liquid forms include solutions, suspensions, and emulsions. In addition to the active ingredient, these preparations may also include coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, cosolvents, etc.

[0239] Oily suspensions can be prepared by suspending the active compound (e.g., a compound of formula (I) or (10b)) in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (e.g., liquid paraffin), or a mixture of these oils. Oily suspensions may contain thickeners, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners may be added to provide a palatable oral preparation, such as glycerol, sorbitol, or sucrose. These preparations may be preserved by the addition of antioxidants, such as ascorbic acid. As an example of an injectable oily carrier, see Minto, J. Pharmacol. Exp. Ther. 281: 93-102, 1997. The pharmaceutical preparations of the present disclosure may also be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil or a mineral oil, as described above, or a mixture of these. Suitable emulsifiers include naturally occurring gums such as gum arabic and gum tragacanth, naturally occurring phospholipids 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, such as in the formulation of syrups and elixirs. Such formulations may also contain analgesics, preservatives or coloring agents.

[0240] The compositions of the present disclosure (including compounds of formula (I) or (10b)) can be delivered by any suitable means, including oral, parenteral and topical methods. Transdermal administration methods via topical routes can be formulated into applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, coatings, powders and aerosols.

[0241] The compositions of the present disclosure (including compounds of formula (I) or (10b)) can also be delivered as microspheres for slow release in vivo. For example, the microspheres can be formulated as drug-containing microspheres for intradermal injection, which are slowly released subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7: 623-645, 1995); as biodegradable and injectable gel formulations (see, e.g., Gao Pharm. Res. 12: 857-863, 1995); or, as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49: 669-674, 1997). Transdermal and intradermal routes can provide continuous administration for weeks or months.

[0242] In another embodiment, the compositions of the present invention can be formulated for parenteral administration, such as intravenous (IV) administration or administration to a body cavity or organ lumen. The preparation for administration generally comprises a solution in which the compositions of the present invention are dissolved in a pharmaceutically acceptable carrier. Acceptable carriers and solvents that can be used include water and Ringer's solution (an isotonic sodium chloride). In addition, sterile fixed oils can be used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids, such as oleic acid, can also be used to prepare injections. These solutions are sterile and generally do not contain unwanted substances. These preparations can be sterilized by various sterilization techniques. As required, the preparations can contain pharmaceutically acceptable auxiliary substances to approach physiological conditions, such as pH regulators and buffers, toxicity regulators, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc. The concentration of the compositions of the present invention in these preparations can vary widely, and will be selected mainly based on fluid volume, viscosity, body weight, etc., according to the selected specific mode of administration and the needs of the patient. For IV administration, the preparation can be a sterile injectable preparation, such as a sterile injectable aqueous or oily suspension. The suspension can be prepared using those suitable dispersants or wetting agents and suspensions. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, such as 1,3-butanediol solution.

[0243] In another embodiment, the preparation of the composition of the present disclosure can be delivered by using liposomes that fuse with the cell membrane or are endocytosed, that is, by using ligands attached to the liposomes or directly attached to the oligonucleotides, such liposomes bind to the surface membrane protein receptors of the cell, resulting in endocytosis. By using liposomes, especially when the liposomes carry ligands specific to the target cells on their surface or otherwise preferentially target specific organs, the composition of the present disclosure can be delivered to the target cells in vivo. (See, for example, Al-Muhammed, J..Microencapsul.13:293-306, 1996; Chonn, Curr.Opin.Biotechnol.6:698-708, 1995; Ostro, Am.J..Ho.sp.Pharm.46:1576-1587, 1989).

[0244] Lipid-based drug delivery systems include lipid solutions, lipid emulsions, lipid dispersions, self-emulsifying drug delivery systems (SEDDS) and self-microemulsifying drug delivery systems (SMEDDS). In particular, SEDDS and SMEDDS are isotropic mixtures of lipids, surfactants and co-surfactants that can spontaneously disperse in aqueous media and form miniemulsions (SEDDS) or microemulsions (SMEDDS). The lipids that can be used in the formulations of the present disclosure include any natural or synthetic lipids, including but not limited to sesame oil, olive oil, castor oil, peanut oil, fatty acid esters, glycerides, and

[0245] The pharmaceutical preparations of the present disclosure can be provided in salt form and can be formed with many acids, including but not limited to hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid, etc. Salts tend to be more soluble in aqueous solvents or other protic solvents as the corresponding free base forms. In other cases, the preparation can be a lyophilized powder in a pH range of 4.5 to 5.5 in, for example, 1 mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol, which is combined with a buffer before use.

[0246] The pharmaceutical preparations of the present disclosure may be provided in salt form and may be formed with bases, i.e., cationic salts, such as alkali metal salts and alkaline earth metal salts, such as sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, and ammonium salts, such as ammonium salts, trimethyl-ammonium salts, diethylammonium salts, and tris-(hydroxymethyl)-methyl-ammonium salts.

[0247] Composition of Sotolacib

[0248] In some embodiments, sotolacib is provided as a tablet comprising sotolacib, microcrystalline cellulose, lactose monohydrate, croscarmellose sodium, and magnesium stearate. In some embodiments, the tablet is film-coated. In some embodiments, the film-coating material comprises polyvinyl alcohol, titanium dioxide, polyethylene glycol, talc, and yellow iron oxide.

[0249] V. Kit

[0250] In another aspect, the present disclosure provides a kit for treating cancer in a subject, the kit comprising:

[0251] a) a therapeutically effective amount of a compound represented by formula (I); and

[0252] b) a therapeutically effective amount of sotolacib,

[0253] and instructions for effective administration,

[0254] wherein the compound of formula (I) is as defined and described herein.

[0255] Cancer and / or solid tumors are described according to Section III-2: Cancer / Solid Tumors. In some embodiments, the cancer and / or solid tumor is any one of the embodiments as described in Section III-2: Cancer / Solid Tumors.

[0256] The subject is described according to Section III-3: Subjects. In some embodiments, the subject is any one of the embodiments as described in Section III-3: Subjects.

[0257] The compound of formula (I) is described according to Section III-1: Compounds of formula (I). In some embodiments, the compound of formula (I) is any of the embodiments described in Section III-1: Compounds of formula (I). In some embodiments, the PTPN11 inhibitor of formula (I) is a compound of formula (10b).

[0258] In some embodiments, the kit includes instructions for administering a compound of formula (I) or (10b) and sotolacib. In some embodiments, the kit includes instructions for administering a compound of formula (10b) and sotolacib. In some embodiments, such instructions include instructions related to safety regulations and the timing and amount of administration of a compound of formula (I) or (10b) and sotolacib. In some embodiments, such instructions include instructions related to safety regulations and the timing and amount of administration of a compound of formula (10b) and sotolacib.

[0259] As described herein, the compound of formula (I) or (10b) and sotolacib can be formulated for concomitant administration or sequential administration. In some embodiments, the compound of formula (I) or (10b) and sotolacib are formulated for concomitant administration. In some embodiments, the compound of formula (I) or (10b) and sotolacib are formulated for sequential administration. In some embodiments, the compound of formula (I) or (10b) is administered before sotolacib is administered. In some embodiments, the compound of formula (I) or (10b) is administered after sotolacib is administered.

[0260] VI. Implementation Methods

[0261] Embodiment 1: A method of treating cancer in a subject, comprising administering to the subject:

[0262] a) a therapeutically effective amount of a compound represented by formula (I):

[0263]

[0264] or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformer, tautomer, or combination thereof; and

[0265] b) a therapeutically effective amount of sotolacib (AMG 510).

[0266] Embodiment 2: The method according to embodiment 1, wherein the compound of formula (I) is represented by formula (10b):

[0267]

[0268] Its name is 6-((3S,4S)-4-amino-3-methyl-2-oxa-8-oxaspiro[4.5]dec-8-yl)-3-(Ra)-(2,3-dichlorophenyl)-2,5-dimethylpyrimidin-4(3H)-one.

[0269] Embodiment 3: The method according to embodiment 1 or embodiment 2, wherein the cancer is characterized by a KRAS mutation.

[0270] Embodiment 4: The method according to embodiment 3, wherein the cancer is characterized by a KRAS G12C mutation.

[0271] Embodiment 5: The method according to any one of embodiments 1 to 4, wherein the cancer comprises a solid tumor.

[0272] Embodiment 6: The method according to any one of embodiments 1 to 5, wherein the cancer is lung cancer, colorectal cancer, pancreatic cancer, urothelial cancer, gastric cancer, mesothelioma, or a combination thereof.

[0273] Embodiment 7: The method according to embodiment 6, wherein the cancer is non-small cell lung cancer (NSCLC).

[0274] Embodiment 8: The method according to any one of embodiments 1 to 7, wherein the cancer is a KRAS G12C-positive cancer that is resistant to a KRAS G12C inhibitor.

[0275] Embodiment 9: The method according to any one of embodiments 1 to 8, wherein the cancer is a KRAS G12C-positive cancer, the cancer being characterized by intrinsic and / or acquired resistance to a KRAS G12C inhibitor.

[0276] Embodiment 10: The method according to any one of embodiments 1 to 9, wherein the cancer is a KRAS G12C-positive cancer that is resistant to sotolacib.

[0277] Embodiment 11: The method according to any one of Embodiments 1 to 10, wherein the cancer progresses or relapses on or after at least one prior line of systemic therapy comprising platinum-based double chemotherapy and / or anti-PD-1 / PD-L1 therapy, each administered as a monotherapy or both administered as a combination therapy.

[0278] Embodiment 12: The method according to any one of embodiments 1 to 11, wherein the subject does not have an activating mutation in BRAFV600X, PTPN11 (SHP2) or KRAS Q61X.

[0279] Embodiment 13: The method according to any one of embodiments 1 to 12, wherein the subject has not been previously treated with a PTPN11 inhibitor.

[0280] Embodiment 14: The method according to any one of embodiments 1 to 12, wherein the subject has been previously treated with a PTPN11 inhibitor other than a compound of formula (I).

[0281] Embodiment 15: The method according to any one of embodiments 1 to 12, wherein the subject has been previously treated with a compound of formula (I).

[0282] Embodiment 16: The method according to any one of embodiments 1 to 8 and 10 to 15, wherein the subject has not been previously treated with a KRAS G12C inhibitor.

[0283] Embodiment 17: The method according to any one of embodiments 1 to 15, wherein the subject has been previously treated with a KRASG12C inhibitor.

[0284] Embodiment 18: The method according to any one of embodiments 1 to 15, wherein the subject has been previously treated with sotolacib.

[0285] Embodiment 19: A method according to any one of embodiments 1 to 18, wherein the subject meets all inclusion criteria according to 1) to 11) of Example 3, provided that the subject does not meet any of the exclusion criteria according to 1) to 17) of Example 3.

[0286] Embodiment 20: The method according to any one of embodiments 1 to 19, wherein the subject is a human.

[0287] Embodiment 21: The method according to any one of embodiments 1 to 20, wherein the compound of formula (I) or (10b) and sotolacib are administered concomitantly.

[0288] Embodiment 22: The method according to any one of embodiments 1 to 20, wherein the compound of formula (I) or (10b) and sotolacib are administered sequentially.

[0289] Embodiment 23: The method according to embodiment 22, wherein the compound of formula (I) or (10b) is administered prior to administration of sotolacib.

[0290] Embodiment 24: The method according to Embodiment 22, wherein the compound of formula (I) or (10b) is administered after administration of sotolacib.

[0291] Embodiment 25: The method according to any one of Embodiments 1 to 24, wherein the compound of formula (I) or (10b) or sotolacib is administered orally, or sotolacib and the compound of formula (I) or (10b) are each administered orally.

[0292] Embodiment 26: The method according to any one of embodiments 1 to 25, wherein the compound of formula (I) or (10b) is administered orally.

[0293] Embodiment 27: The method according to any one of embodiments 1 to 26, wherein sotolacib is administered orally.

[0294] Embodiment 28: The method according to any one of embodiments 1 to 27, wherein the compound of formula (I) or (10b) and sotolacib are provided in a combined therapeutically effective amount.

[0295] Embodiment 29: The method according to any one of embodiments 1 to 27, wherein the compound of formula (I) or (10b) and sotolacib are provided in synergistically effective amounts.

[0296] Embodiment 30: The method according to any one of embodiments 1 to 29, wherein the compound of formula (I) or (10b) and / or sotolacib are used in a dose different from when they are used alone.

[0297] Embodiment 31: The method according to embodiment 30, wherein the compound of formula (I) or (10b) is used in a dose lower than when it is used alone.

[0298] Embodiment 32: The method according to embodiment 30, wherein the compound of formula (I) or (10b) is used in a dose higher than when it is used alone.

[0299] Embodiment 33: The method according to any one of embodiments 30 to 32, wherein sotolacib is used in a lower dose than when it is used alone.

[0300] Embodiment 34: The method according to any one of embodiments 30 to 32, wherein sotolacib is used in a higher dose than when it is used alone.

[0301] Embodiment 35: A method according to any one of embodiments 1 to 34, wherein the treatment comprises one or more treatment cycles; wherein each of the one or more treatment cycles has a duration of about 28 days; and the compound of formula (I) or (10b) and / or sotolacib is administered daily.

[0302] Embodiment 36: The method according to any one of Embodiments 1 to 35, wherein the administration of the compound of Formula (I) or (10b) and sotolacib comprises one or more dose escalations, dose maintenance, or dose reductions of the compound of Formula (I) or (10b) and / or sotolacib.

[0303] Embodiment 37: The method according to Embodiment 36, wherein the administration of the compound of Formula (I) or (10b) and sotolacib comprises one or more dose escalations, dose maintenance, or dose reductions of the compound of Formula (I) or (10b).

[0304] Embodiment 38: The method according to Embodiment 37, wherein one or more dose escalations, dose holds, or dose reductions of the compound of formula (I) or (10b) are determined by dose limiting toxicity (DLT) assessment.

[0305] Embodiment 39: The method according to Embodiment 38, wherein administration of the compound of Formula (I) or (10b) comprises dose escalation following the previous treatment cycle when the dose limiting toxicity (DLT) rate is less than about 19.7% as determined by DLT assessment.

[0306] Embodiment 40: The method according to Embodiment 38, wherein administration of the compound of formula (I) or (10b) comprises dose reduction following the previous treatment cycle when the dose limiting toxicity rate is greater than about 29.8% as determined by DLT assessment.

[0307] Embodiment 41: The method according to embodiment 38, wherein administration of the compound of formula (I) or (10b) comprises dose maintenance following the previous treatment cycle when the dose limiting toxicity rate is in the range of about 21.9% to about 29.8% as determined by DLT assessment.

[0308] Embodiment 42: A method according to any one of embodiments 35 to 41, wherein the treatment comprises a dose escalation phase, and wherein after the dose escalation phase, the treatment further comprises a dose expansion / optimization phase; and the compound of formula (I) or (10b) is administered at a dosage regimen determined during the dose escalation phase.

[0309] Embodiment 43: The method according to Embodiment 42, wherein during the dose expansion / optimization phase, administration of the compound of formula (I) or (10b) comprises one or more dose adjustments.

[0310] Embodiment 44: The method according to any one of embodiments 1 to 43, wherein the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg or about 960 mg.

[0311] Embodiment 45: The method according to embodiment 44, wherein the therapeutically effective amount of sotolacib is a total daily dose of about 960 mg.

[0312] Embodiment 46: A method according to any one of embodiments 1 to 45, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 450 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 550 mg to about 1000 mg, from about 600 mg to about 1000 mg, from about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 100 mg to about 700 mg, from about 150 mg to about 700 mg, from about 200 mg to about 700 mg, from about from about 250 mg to about 700 mg, from about 300 mg to about 700 mg, from about 3 mg to about 700 mg, from about 400 mg to about 700 mg, from about 450 mg to about 700 mg, from about 500 mg to about 700 mg, from about 550 mg to about 700 mg, from about 100 mg to about 550 mg, from about 150 mg to about 550 mg, from about 200 mg to about 550 mg, from about 250 mg to about 500 mg, from about 250 mg to about 500 mg, from about 300 mg to about 700 mg, from about 400 mg to about 700 mg, from about 450 mg to about 700 mg, from about 500 mg to about 700 mg, from about 550 mg to about 700 mg, from about g to about 550 mg, from about 300 mg to about 550 mg, from about 350 mg to about 550 mg, from about 400 mg to about 550 mg, from about 450 mg to about 550 mg, from about 100 mg to about 400 mg, from about 150 mg to about 400 mg, from about 200 mg to about 400 mg, from about 250 mg to about 400 mg, or from about 300 mg to about 400 mg, on a salt-free and water-free basis.

[0313] Embodiment 47: A method according to embodiment 46, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg, on a salt-free and water-free basis.

[0314] Embodiment 48: The method according to Embodiment 47, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of about 250 mg, about 400 mg or about 550 mg, calculated on a salt-free and water-free basis.

[0315] Embodiment 49: The method according to any one of embodiments 46 to 48, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of about 250 mg, calculated on a salt-free and water-free basis.

[0316] Embodiment 50: The method according to any one of embodiments 46 to 48, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of about 400 mg, calculated on a salt-free and water-free basis.

[0317] Embodiment 51: The method according to any one of Embodiments 46 to 48, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is a total daily dose of about 550 mg, calculated on a salt-free and water-free basis.

[0318] Embodiment 52: The method according to any one of Embodiments 1 to 51, wherein the compound of formula (10b) and sotolacib are each administered orally.

[0319] Embodiment 53: The method according to any one of embodiments 1 to 52, wherein the compound of formula (10b) is administered once, twice, three times or four times daily.

[0320] Embodiment 54: The method according to embodiment 53, wherein the compound of formula (I) or (10b) is administered once daily; and sotolacib is administered once daily.

[0321] Embodiment 55: The method according to any one of embodiments 1 to 54, wherein the compound of formula (I) or (10b) is provided in a tablet formulation.

[0322] Embodiment 56: The method of any one of embodiments 1 to 55, wherein the treatment reduces the volume of the cancer or solid tumor by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90%.

[0323] Embodiment 57: The method according to any one of embodiments 1 to 55, wherein the treatment is of a stable cancer or a solid tumor.

[0324] Embodiment 58: The method according to any one of embodiments 1 to 57, wherein the subject is further evaluated for one or more biomarkers associated with anti-tumor response.

[0325] Embodiment 59: A kit for treating cancer in a subject, comprising:

[0326] a) a therapeutically effective amount of a compound represented by formula (I):

[0327]

[0328] or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformational isomer, tautomer or a combination thereof, or a compound represented by formula (10b):

[0329] and

[0330] b) a therapeutically effective amount of sotolacib,

[0331] and instructions for effective administration.

[0332] Embodiment 60: The kit according to embodiment 59, wherein the compound of formula (I) or (10b) and sotolacib are formulated for concomitant administration.

[0333] Embodiment 61: The kit according to embodiment 59, wherein the compound of formula (I) or (10b) and sotolacib are formulated for sequential administration.

[0334] VII. List of Abbreviations

[0335]

[0336]

[0337]

[0338]

[0339] VIII. Examples

[0340] Example 1: In vivo effectiveness of formula (10b) and compound A

[0341] A. Materials

[0342] Test object #1 - Formula (10b):

[0343] Chemical name: 6-[(3S, 4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]dec-8-yl]-3-(2,3-dichlorophenyl)-2,5-dimethyl-3,4-dihydropyrimidin-4-one

[0344] Molecular formula: C 21 H 26 Cl2N4O2

[0345] Molecular weight: 437.37

[0346] Test #2 - Compound A:

[0347] Chemical name: 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one

[0348] CAS#: 252403-56-6

[0349] Molecular formula: C 30 H 30 F2N6O3

[0350] Molecular weight: 560.6

[0351] Cell line: NCI-H358 cell line was purchased from ATCC. It is a human NSCLC cell line carrying a heterozygous mutation of KRAS G12C. Cells were cultured in RPMI 1640 containing glutamine (ThermoFisher #22400-089) + 10% fetal bovine serum (FBS, Thermo Fisher #10099-141) in a 37°C tissue culture incubator (Thermo Fisher) containing 5% CO2.

[0352] Test animals: Female NOD / SCID mice (Beijing Anikeeper Biotech Co., Ltd) were used in this experiment. The animals were 6 to 8 weeks old at the time of xenograft implantation. The protocol involving the care and use of animals in this study and any amendments or procedures were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Crown Bioscience Ltd. before implementation. During the study, animal care and use were performed in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC).

[0353] All mice were housed in an AAALAC-accredited animal research facility at the Light Muller Building, Changping Sector of Zhongguancun Scientific Park, No. 21 Huoju Road, Changping District, Beijing, China. All animals were maintained under veterinary supervision and care at an animal facility that oversees a comprehensive and well-executed health surveillance program.

[0354] Mice were housed in polysulfone IVC cages (325 mm × 210 mm × 180 mm) with a maximum of five animals, free access to food and water, and 12-hour light and dark alternation. All animals were fed with irradiated standard rodent diet-rat and mouse growth and breeding feed (Beijing KeaoXieli Feed Co., LTD) freely. Mice were monitored daily and cages were changed every other week.

[0355] B. Experimental Procedure

[0356] Formulation: Formula (10b) was prepared in 0.5% methylcellulose. To prepare a 0.5% methylcellulose solution, methylcellulose powder (400 cP, Sigma #M0262) was added to heated 80°C sterile water. The solution was incubated at 80°C with stirring for 3-4 hours, followed by incubation at 4°C with continuous stirring for 18 hours. After adjusting the final volume with sterile H2O, the solution was stirred for another 30 min at 4°C and then filtered using a 0.45 μM sterile filter. The prepared 0.5% methylcellulose solution was stored at 4°C for future use.

[0357] In order to prepare the administration suspension of formula (10b) in 0.5% methylcellulose, the compound weighed is placed in a glass vial, and 0.5% methylcellulose solution is added to the vial with a syringe. After vortexing for 30 seconds, the vial is ultrasonicated for about 20 minutes at room temperature on a "high" setting in a water bath sonicator (Shanghai Kudos Ultrasonic Instrument Co., LTD, model SK2510HP), until an off-white suspension without visible solids is obtained. The administration suspension prepared is stored at 4 ° C, while stirring continuously and gently. Fresh administration suspension is prepared once a week.

[0358] Compound A was prepared in Labrasol (Gattefosse #3074). To prepare the Compound A dosing solution, the weighed compound was placed in a glass vial, and Labrasol was added to the vial. The mixture was vortexed for 30 seconds and sonicated for about 5 min in a water bath sonicator (Shanghai Kudos Ultrasonic Instrument Co., LTD, model SK2510HP) until a light yellow solution was obtained. The dosing solution was stored at 4 ° C while stirring gently and continuously. Fresh dosing solutions were prepared once a week. Immediately before the administration of Compound A to the animals, the dosing solution was mixed with an equal volume of sterile ddH2O, vortexed, and then incubated at room temperature for 10 min, at which time a uniform solution was formed and used for dosing mice. The diluted dosing solution was used within one hour after adding water.

[0359] In vivo modeling, treatment and data analysis: For xenograft studies with NCI-H358 cells, 5 million cells with 50% Matrigel (BD bioscience #356234) in sterile PBS (Hyclone Laboratories #SH30256.01) were injected subcutaneously into the right flank of female 6-8 week old NOD / SCID mice (Beijing Anikeeper Biotech Co., Ltd). Tumor size was measured with a caliper and the standard formula was used: length x width 2 / 2, where length and width are the long and short diameters of the tumor, respectively.

[0360] When the average tumor volume reaches 250 mm 3 When, based on tumor volume and body weight, mice are randomly grouped into 8 groups. " Distribution of Matching " method / " Stratification " method based on StudyDirector software version 3.1.399.19 (StudyLog) is randomly grouped. Then, as shown below, by oral gavage (PO), mice vehicle, a single compound under one of three dosing levels (100mg / kg QD, 30mg / kg QD and 10mg / kg QD), a single formula (10b) or a combination of two compounds under one of two dosing levels (100mg / kg QD and 50mg / kg QD) are used. Formula (10b) is used in the morning and compound A is used in the afternoon, wherein the morning and afternoon dosing intervals are 6 hours.

[0361] Group treat A 0.5% Methylcellulose QD AM + 50% Labrasol QD PM B 0.5% methylcellulose AM + compound A 100 mg / kg QD PM C 0.5% methylcellulose AM + compound A 30 mg / kg QD PM D 0.5% methylcellulose AM + compound A 10 mg / kg QD PM E Formula (10b) 100mg / kg QD AM+50% Labrasol QD PM F Formula (10b)50mg / kg QD AM+50% Labrasol QD PM G Formula (10b) 100 mg / kg QD AM + Compound A 100 mg / kg QD PM H Formula (10b) 50 mg / kg QD AM + Compound A 100 mg / kg QD PM I Formula (10b) 100 mg / kg QD AM + Compound A 30 mg / kg QD PM J Formula (10b) 100 mg / kg QD AM + Compound A 10 mg / kg QD PM

[0362] Dosing was started about 2 weeks after subcutaneous implantation of tumor cells. The dosing volume of formula (10b) is 5mL / kg, and the dosing volume of compound A is 6mL / kg, and is adjusted based on the weight of a single mouse measured once every two weeks. Tumor volume was measured twice a week. Body weight and tumor volume were collected using StudyDirector software version 3.1.399.19 (StudyLog). 21 days after administration (all dosing levels, except 10mg / kg formula (10b)) or 29 days (10mg / kg formula (10b)), according to the blood collection method described below, 2 hours after the last dose of compound A (8 hours after the last dose of formula (10b)) plasma samples were collected.

[0363] Data were analyzed using Microsoft Excel and GraphPad Prism 8.0. Day 0 was the day before treatment started. Body weight change and tumor growth inhibition (TGI) were calculated based on the following formulas.

[0364] Weight change% = (BW i -BW0) / BW0*100%

[0365] BW i BW and BW0 are the body weights of individual mice on measurement day 1 and day 0, respectively.

[0366] TGI%=(C i -T i ) / (C i -C0)*100%

[0367] T i and C i are the mean tumor volumes of the treatment group and the vehicle group on the measurement day, respectively; C0 is the mean tumor volume of the vehicle group on day 0.

[0368] Blood collection for pharmacokinetic analysis: Blood from live animals was obtained through the retroorbital sinus for pharmacokinetic (PK) analysis. The mouse was tightly grasped behind the ear and gently rotated to one side so that a disposable sterile capillary was inserted behind the eyeball and gently rotated to destroy the sinus cavity. The blood flowed through the capillary and was directly added to a volume of 100 μL in a K2EDTA tube (Jiangsu KangJianMedical Apparatus Corporation #044022). The blood in the tube was inverted several times to evenly distribute the EDTA. In order to separate plasma, each tube was centrifuged at 8000 rpm for 5 minutes without braking in a desktop centrifuge (Eppendorf, model 5424R). The supernatant (plasma) was carefully transferred to a microcentrifuge tube and placed on dry ice before storage at -80°C.

[0369] LC-MS / MS quantification of Formula (10b) and Compound A in mouse plasma: The concentrations of Formula (10b) and Compound A in mouse plasma were quantified using a validated LC-MS / MS method. For plasma sample analysis, 20 μL of each sample was precipitated with 200 μL of acetonitrile containing glipizide (100 ng / mL) as an internal standard. The suspension was vortexed for 1 min and centrifuged at 4,000 rpm for 10 min, after which 80 μL of the extract was aliquoted for LC-MS / MS analysis. The HPLC-MS / MS was performed on a Shimadzu Nexera UHPLC system in positive mode (ESI +LC-MS / MS analysis was performed on a Sciex 6500TQ-S triple quadrupole mass spectrometer (MS / MS) operated under ). The mass spectrometry source conditions were set as follows: ion spray voltage (5500 V), CAD (8), CUR (35), TEM (450), GS1 (60), GS2 (60), EP (10), CXP (12), CEM (2000); and for formula (10b): DP (40), CE (65), for compound A: DP (40), CE (47), for internal standard: DP (35), CE (20). Formula (10b) and compound A were separated using a Waters X-Bridge BEH C18 column (2.1×50 mm, 1.7 μm) and detected by multiple reaction monitoring transitions (m / z 437.10>186.10 for formula (10b), m / z 561.20>134.20 for compound A, and m / z 446.20>321.10 for internal standard). The injection volume was 2 uL. LC mobile phase A was 0.025% formic acid-water containing 1 mM ammonium acetate, and mobile phase B was 100% methanol containing 5 mM ammonium acetate. The gradient (% B) was 2% (0-0.4 min), 2-65% (0.4-1.7 min), 65-90% (0.7 to 1.3 min), 90% (1.3-1.9 min), 90-2% (1.9 to 1.91 min), 2% (1.91-2.5 min). The flow rate was 0.6 mL / min. The column temperature was 60° C. Under these conditions, the retention time for formula (10b) was 1.42 min, and the retention time for compound A and internal standard was 1.44 min. The method was validated in untreated CD-1 mouse plasma with an analytical range of 3-3,000 nM (for both formula (10b) and compound A).

[0370] C. Results

[0371] Sotolacib (AMG 510) is a KRAS covalent inhibitor that specifically targets the KRAS G12C mutant. It has demonstrated single-agent responses in patients with solid tumors (including NSCLC) carrying the KRAS G12C mutation. In particular, in the CodeBreaK 100 study, the subgroup with NSCLC had a confirmed 32.2% objective response rate (complete or partial response) and 88.1% disease control rate (objective response or stable disease); the median progression-free survival was 6.3 months. Hong et al., N. Engl. J. Med. 2020; 383: 1207-1217. Unbiased functional genomics approaches also identified SHP2 as a weakness under KRASG12C inhibition. KRAS G12C inhibition induces adaptive feedback activation of the MAPK pathway through multiple RTKs, which signal through SHP2. This study was designed to evaluate the effect of the combination of the SHP2 inhibitor formula (10b) and the KRAS G12C inhibitor Compound A in an in vivo human NSCLC tumor model bearing KRAS G12C.

[0372] Combination treatment with formula (10b) and compound A resulted in tumor growth inhibition in a subcutaneous model of NSCLC carrying KRAS G12C: A mouse tumor xenograft model was generated by subcutaneous implantation of NCI-H358 cells carrying KRASG12C, and was used to test the in vivo anti-tumor response of the combination of formula (10b) and compound A. Mice carrying established NCI-H358 subcutaneous tumors were randomly divided into groups and treated with vehicle, compound A alone, formula (10b) alone, or a combination of the two compounds, delivered orally for 21 days. Formula (10b) was administered daily in the morning, and compound A was administered daily in the afternoon, with a 6-hour interval between morning and afternoon dosing. Tumor volume was monitored by caliper once every two weeks, and body weight was recorded. Figures 1A to 1E The data shown in are expressed as mean ± SEM, and for Figure 1B-1E , N = 8 mice / group and for Figure 1A , N=10 mice / group.

[0373] Figure 1A Shown are the mean (+ / -SEM) tumor volumes (mm) of female NOD / SCID mice bearing NCI-H358 tumors following daily oral administration of Formula (10b), Compound A, or a combination of the two test articles from Day 1 to Day 28 at the indicated dose levels. 3 ).

[0374] like Figure 1A , 1BAs shown in Figures 1 and 1D and Tables 1 and 2, treatment with formula (10b) alone inhibited the growth of NCI-H358 tumors in a dose-dependent manner, resulting in 89% and 46% tumor growth inhibition (TGI) at 100 mg / kg QD and 50 mg / kg QD, respectively, on day 21 of the study. Treatment with compound A alone also inhibited NCI-H358 tumor growth in a dose-dependent manner, with tumor regression observed at 100 mg / kg QD, 30 mg / kg QD, and 10 mg / kg QD. Notably, the combination of compound A 10 mg / kg QD and formula (10b) 100 mg / kg QD more potently inhibited tumor growth than single agents, causing tumor regression ( Figure 1A ). It is worth noting that when formula (10b) 100 mg / kg QD is combined with compound A at 30 mg / kg and 100 mg / kg QD ( Figure 1B and Figure 1D ), a significant enhancement of antitumor activity was observed, which may be attributed to the robust single-agent response of Compound A. All dosing conditions were considered well tolerated, as shown by the maintenance of mouse body weight during the study ( Figure 1C and Figure 1E Although random mouse losses were observed in this study, they were not considered treatment-related due to the lack of dose-dependency (Table 3).

[0375] Table 1: Summary of combination studies of formula (10b) at 100 mg / kg and Compound A at 10 mg / kg.

[0376]

[0377] Table 2: From Figure 1B-1E For the study shown in , endpoint TGI and plasma concentrations at 2 hours after the last dose of Compound A (8 hours after the last dose of Formula (10b)).

[0378]

[0379]

[0380] TGI% = (Ci-Ti) / (Ci-C0) * 100%. Ti and Ci are the average tumor volumes of the treatment group and the vehicle group on the measurement day, respectively; C0 represents the average tumor volume of the vehicle group on day 0. For plasma concentrations, data are expressed as mean ± SD. N = 4 mice / group. BQL, below the limit of quantitation (3 nM).

[0381] Pharmacokinetic analysis was performed 2 hours after the last dose of Compound A (8 hours after the last dose of Formula (10b)) (Table 2). 2 hours after the last dose, significant concentrations of Compound A were detected in plasma, with 30 mg / kg producing 2.0 μM and 100 mg / kg producing 8.8 μM. Compound A plasma concentrations were not affected by treatment with Formula (10b). When administered as a single agent (7.5 μM) and in combination with Compound A (5-10 μM), Formula (10b) 100 mg / kg produced similar significant plasma concentrations 8 hours after administration. When administered as a single agent (4.4 μM) and in combination with Compound A (5.5 μM), administration of Formula (10b) 50 mg / kg also produced similar significant plasma concentrations 8 hours after administration. The data indicate that there is no drug-drug interaction between Compound A and Formula (10b) in mice.

[0382] Table 3: From Figure 1B-1E An overview of the mouse morbidity in the pharmacodynamic studies is shown in .

[0383]

[0384] Each event (mouse loss) indicates the loss of one mouse, where the day refers to the dosing day when the mouse was removed from the study. BWL, body weight loss; QD, dosing once daily.

[0385] D. Conclusion

[0386] In the NCI-H358 NSCLC xenograft model carrying the KRAS G12C mutation, combined treatment with formula (10b) and compound A robustly inhibited tumor growth. In detail, treatment with compound A 10 mg / kg QD as a single agent caused tumor growth inhibition, and the combination with formula (10b) caused tumor regression. Treatment with higher doses of compound A (30 mg / kg QD or 100 mg / kg QD) as a single agent caused robust tumor regression; the combination with formula (10b) did not significantly further inhibit tumor volume. Treatment with formula (10b) monotherapy at 100 mg / kg QD caused approximate tumor stagnation. All tested treatment conditions were well tolerated in the mouse model.

[0387] Example 2: In vivo effectiveness of the combination of formula (10b) and sotolacib

[0388] A. Materials

[0389] Test object #1 - Formula (10b):

[0390] Chemical name: 6-[(3S, 4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]dec-8-yl]-3-(2,3-dichlorophenyl)-2,5-dimethyl-3,4-dihydropyrimidin-4-one

[0391] Molecular formula: C 21 H 26 Cl2N4O2

[0392] Molecular weight: 437.37

[0393] Test Article #2 - Sotolacib:

[0394] CAS#: 2296729-00-3

[0395] Molecular formula: C 30 H 30 F2N6O3

[0396] Molecular weight: 560.6

[0397] B. Experimental Procedure

[0398] Tumor cell line culture: Human NSCLC NCI-H358 cell line (Shanghai Institutes for Biological Sciences of Chinese Academy of Sciences) was maintained in vitro in a 37°C incubator with 5% CO2 using RPMI-1640 medium (Gibco, catalog number C22400500BT) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, catalog number 10099-141C). NCI-H358 cells at passage 9 were thawed and expanded, and cells at passage 13 were collected for inoculation. Human NSCLC NCI-H2122 cell line (American Type Culture Collection) was maintained in vitro in a 37°C incubator with 5% CO2 using RPMI-1640 medium (Gibco, catalog number C22400500BT) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, catalog number 10091-148). NCI-H2122 cells at passage 6 were thawed and expanded, and cells at passage 14 were collected for plating.

[0399] Animal inoculation and randomization: All experiments were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Crown Bioscience, Inc. before they were performed and were performed in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). For the NCI-H358 study, 75 6- to 8-week-old NOD / SCID female mice (Beijing Anikeeper Biotech Co., Ltd) were subcutaneously inoculated with NCI-H358 tumor cells (5×10 6 These tumor cells were suspended in PBS:Matrigel (Corning, catalog number 354234) at a ratio of 1:1 in a volume of 0.1 mL / mouse. At 13 days after cell inoculation, when the average tumor volume was 160 mm 3 According to the tumor volume (size mm 3 ), the mice were randomly divided into four groups (10 mice / group). For the NCI-H2122 study, 80 6- to 8-week-old BALB / c female nude mice (GemPharmatech Co., Ltd) were subcutaneously inoculated with NCI-H2122 tumor cells (1x10 7 These tumor cells were suspended in PBS:Matrigel (Corning, catalog number 354234) at a ratio of 1:1, with a volume of 0.1 mL / mouse. Nine days after cell inoculation, when the average tumor volume was 180 mm 3 When the tumor volume (size mm 3 ), and the mice were randomly divided into four groups (10 mice / group).

[0400] Dosing: Four groups of female NOD / SCID mice (10 animals / group) bearing NCI-H358 tumors were administered one of the following treatment regimens by oral gavage once daily for a period of 28 days: 1) vehicle, 2) Formula (10b) (100 mg / kg), 3) sotolacib (10 mg / kg), or 4) a combination of Formula (10b) (100 mg / kg) and sotolacib (10 mg / kg). Four groups of female BALB / c nude mice (10 animals / group) bearing NCI-H2122 tumors were administered one of the following treatment regimens by oral gavage once daily for a period of 21 days: 1) vehicle, 2) Formula (10b) (100 mg / kg), 3) sotolacib (100 mg / kg), or 4) a combination of Formula (10b) (100 mg / kg) and sotolacib (100 mg / kg).

[0401] Formulation of Formula (10b): The formulation of Formula (10b) buffer (0.5% v / v methylcellulose in sterile deionized water) was prepared by weighing the required amount of methylcellulose (Sigma-Aldrich, catalog number M0262, viscosity 400 cP) into a glass bottle. Sterile deionized water equivalent to 75% v / v of the expected final volume was added under continuous magnetic stirring with a stir bar and stirred at room temperature until completely dissolved. The buffer was then brought to final volume with anhydrous deionized water. A correction factor of 1.025 was applied to the formulation of Formula (10b) to accommodate the purity (97.7%).

[0402] A 10 mg / mL (10.25 mg / mL in the case of a correction factor) working suspension of formula (10b) was prepared and a 100 mg / kg dose was administered to mice by oral gavage at a 10 mL / kg dose volume. To prepare the formula (10b) formulation, the compound was accurately weighed into a glass bottle. The formula (10b) formulation buffer equivalent to 70% v / v of the expected final volume was added to the glass bottle containing the bulk drug and fully mixed for 4 to 9 minutes using a 1 / 4 inch probe until a homogeneous suspension without larger visible aggregates / particles was obtained. The remaining portion of the suspension vehicle was added to the dispersion containing the bulk drug to reach the expected final volume. The suspension was fully mixed for 30 minutes, stored at 4°C for one week, and fully mixed before daily administration and throughout the administration period.

[0403] Sotolacib Formulation: Sotolacib formulation buffer (2% v / v HPMC / 1% v / v Tween 80 in sterile deionized water) was prepared by weighing the required amount of HPMC (Sigma-Aldrich, catalog number H3785) and Tween 80 (Sigma-Aldrich, catalog number P4780) into a glass bottle. Sterile deionized water equivalent to 80% v / v of the expected final volume was added under continuous magnetic stirring with a stir bar and stirred at room temperature until completely dissolved. Then, the buffer was brought to final volume with sterile deionized water.

[0404] 1mg / mL and 10mg / mL of Sotolacib working suspensions were prepared and 10mg / kg and 100mg / kg doses were administered to mice by oral gavage at a 10mL / kg dose volume. To prepare the Sotolacib formulation, the compound was accurately weighed into a glass bottle. The Sotolacib formulation buffer equivalent to the expected final volume of 100% v / v was added to the glass bottle containing the drug substance and mixed thoroughly until a homogeneous suspension without larger visible aggregates / particles was obtained. The suspension was mixed thoroughly and the working suspension was administered as quickly as possible. The suspension was prepared daily and mixed thoroughly before administration and throughout the administration period.

[0405] Tumor and body weight measurements: Tumor volume was measured twice weekly in two dimensions using calipers after randomization, and volume was expressed in mm using the formula 3 Mean: tumor volume = (length x width x width) / 2, wherein length is defined as the longest tumor dimension, and width is defined as the longest tumor dimension perpendicular to the length. Dosing and tumor and body weight measurements were performed in a fume hood. The tumor volume of individual mice in each group (and the average tumor volume (± standard error of the mean (SEM))) were recorded. The tumor growth inhibition (TGI) (indicator of the anti-tumor effect) of each treatment group (T) and control group (C) was calculated using the average tumor volume measurements on day 1 (0) and the indicated measurement day (i), using the formula: TGI (%) = (1-(Ti-T0) / (Ci-C0)) x100. TGI was calculated on the day after the last continuous daily dose (day 29) and reported only when ≤100%. Tumor regression (REG) was also assessed and defined as a tumor with a smaller tumor volume on the indicated study day compared to the first dose on day 1. If TGI>100%, mean tumor regression is reported instead and was calculated by determining the mean percentage of tumor regression on the day following the last consecutive daily dose as indicated compared to the first dose on Day 1.

[0406] Body weight (BW) was also recorded twice a week. Using the body weight on day 1 (0) of the first day of dosing and the body weight on the indicated (i) day after dosing, the following formula was used: BW change (%) = ((BWi / BW0) × 100)-100 to calculate the individual mouse body weight and body weight change expressed as a percentage for each animal. The average body weight change (± SEM) was recorded by group. If >20% of the mice in the group had ≥20% weight loss, or >20% of the mice in the group died spontaneously or had any painful clinical symptoms requiring euthanasia, the treatment was considered to be intolerant.

[0407] Record animal tumor and body weight measurements and use Study Director TM All analyses were performed using GraphPad Prism software (version 9) or calculated from raw data in Microsoft Excel.

[0408] Statistical analysis: Statistical analysis was performed using GraphPad Prism software (version 9). For statistical analysis comparing the vehicle group with all other groups, a two-way repeated measures analysis of variance (ANOVA) was applied within the specified days, followed by a post hoc Tukey multiple comparison test for the mean. For statistical analysis comparing monotherapy and combination groups, a two-way repeated measures ANOVA was performed between the mean of each monotherapy group and the mean of the combination group within the specified days. A p value of less than 0.05 was considered statistically significant.

[0409] Experimental endpoints: Animal health was monitored at least twice a week by behavioral observation and body weight examination. Animals were examined daily for morbidity and mortality following tumor inoculation. During routine monitoring, animals were examined for tumor growth and any effects of treatment on behavior, such as motility, food and water consumption, weight gain / loss, eye / hair roughness, and any other abnormalities.

[0410] The study endpoint was defined as when an animal reached one of the following euthanasia criteria: ≥20% body weight loss relative to day 1 at any time; ≥15% body weight loss relative to day 1 for 72 hours; single tumor volume ≥3000mm 3 , or when the mean tumor volume reaches ≥2000mm 3 All mice in each group were euthanized at 4 dpi. In the NCI-H358 study, animals in all groups were euthanized on day 34 or 35, and in the NCI-H2122 study, animals in all groups were euthanized on day 21 or 22.

[0411] C. Results

[0412] Sotolacib (AMG 510) is a KRAS covalent inhibitor that specifically targets the KRAS G12C mutant. It has demonstrated single-agent responses in patients with solid tumors (including NSCLC) carrying KRAS G12C mutations. In particular, in the CodeBreaK 100 study, the subgroup with NSCLC had a confirmed 32.2% objective response rate (complete or partial response) and 88.1% disease control rate (objective response or stable disease); the median progression-free survival was 6.3 months. Hong et al., N. Engl. J. Med. 2020; 383: 1207-1217. Cell line studies have shown that SHP2 inhibitors increase KRAS-GDP occupancy and enhance the effectiveness of KRAS G12C inhibitors because the current generation of KRASG12C inhibitors only inhibits the GDP-bound form of KRAS G12C. Unbiased functional genomics approaches also identified SHP2 as a weakness under KRAS G12C inhibition. Adding SHP2 inhibitors to therapeutics targeting mutant KRAS G12C can improve effectiveness in several ways. Because mutant KRASG12C maintains a certain degree of cycling between these two states, the GTP-bound form of KRAS G12C can still activate ERK signaling in the presence of sotolacib. SHP2 inhibition reduces the SOS1-dependent GTP load of RAS, which causes elevated RAS-GDP levels and can therefore enhance the effectiveness of sotolacib. In addition, compensatory bypass signaling feedback activation mediated upstream or downstream of the RTK / MAPK pathway has been shown to reduce the effectiveness of KRASG12C inhibitors preclinically and clinically, and SHP2 inhibitors can inhibit this feedback activation.

[0413] SHP2 inhibition may also enhance the antitumor activity of KRAS G12C inhibitors in patients with molecular subtypes with particularly poor outcomes. For example, mutations in the tumor suppressor Kelch-like ECH-associated protein 1 (KEAP 1) are found in approximately 20% of KRAS-mutant SCLCs, and co-mutations of KRAS and KEAP1 are associated with poor prognosis and clinical outcomes.

[0414] Two studies were conducted to determine whether the combination of the SHP2 inhibitor formula (10b) and sotolacib enhanced antitumor activity compared to either agent alone. Studies were conducted using a xenograft model derived from the NSCLC NCI-H358 cell line carrying a KRAS G12C mutation and a xenograft model derived from the NSCLC NCI-H2122 cell line carrying a co-mutation of KRASG12C, KEAP1, and STK11.

[0415] Combination treatment with formula (10b) and sotolacib caused tumor regression in the NCI-H358 subcutaneous xenograft model of NSCLC carrying the KRAS G12C mutation. An in vivo study was conducted to evaluate the effect of the combination of formula (10b) and the KRASG12C inhibitor sotolacib on tumor volume in female NOD / SCID mice carrying the NCI-H358 NSCLC xenograft model derived from a cell line carrying the KRASG12C mutation. Female NOD / SCID mice were subcutaneously implanted with NSCLC NCI-H358 tumor cells. When the tumor reached 60 mm 3 When the average size of the tumor was 100 mg / kg, the mice were randomly divided into treatment groups (n=10 / group) and administered the indicated levels of vehicle, 100 mg / kg formula (10b), 10 mg / kg sotolacib, or a combination of 100 mg / kg formula (10b) and 100 mg / kg sotolacib by oral gavage once a day from day 1 to day 28. The tumor volume of the mice was measured twice a week until day 34, and the results are shown until day 29.

[0416] like Figure 2A As shown in Table 4, the combination of Formula (10b) and Sotolacib showed significantly greater antitumor activity than any of the test subjects as monotherapy. After oral administration of the combination of Formula (10b) (100 mg / kg) and Sotolacib (10 mg / kg) once daily, compared to the two monotherapy groups ( Figure 2A and Table 4), a statistically significant reduction in tumor volume was observed in the combination group, and the average tumor regression on day 29 was significantly increased to 83% (Table 4). The results also showed that both formula (10b) and sotolacib had monotherapy antitumor activity at the tested dose levels. After oral administration of formula (10b) (100 mg / kg) or sotolacib (10 mg / kg) once daily, a statistically significant reduction in tumor volume was observed in both treatment groups compared to the vehicle group ( Figure 2A and Table 4), where the mean tumor regressions on day 29 in the formula (10b) and sotolacib monotherapy groups were 14% and 13%, respectively.

[0417] Table 4: Summary of the antitumor activity of Formula (10b) and Sotolacib as monotherapy and in combination in NOD / SCID mice bearing tumors derived from the NCI-H358 subcutaneous cell line.

[0418]

[0419] Abbreviations: ANOVA = analysis of variance; NOD / SCID = non-obese diabetes / severe combined immunodeficiency; n = number; REG = regression; SEM = standard error of the mean.

[0420] Note: Oral dosing was performed daily from day 1 to day 28, and the last tumor measurement was performed on day 34. For statistical analysis, two-way repeated measures ANOVA of group means followed by post hoc Tukey's multiple comparison test was performed from day 5 to day 29 for vehicle group comparison*, and two-way repeated measures ANOVA of monotherapy and combination group means was performed from day 5 to day 29 for combination group comparison**.

[0421] like Figure 2B As shown, in the mouse model, treatment had no effect on body weight and all treatments were well tolerated.

[0422] Combination treatment with formula (10b) and sotolacib caused tumor growth inhibition in the NCI-H2122 subcutaneous xenograft model of NSCLC carrying KRAS G12C, KEAP1 and STK11 co-mutations. In vivo studies were conducted to evaluate the effect of the combination of formula (10b) and the KRASG12C inhibitor sotolacib on tumor volume in BALB / c nude mice in the NCI-H2122 NSCLC xenograft model derived from a cell line carrying KRAS G12C mutations. The model also has several coexisting mutations, including frameshift insertions in KEAP1 (KEAP1A203fs) and frameshift deletions in STK11 (STK11G279fs) that cause loss of function of KEAP1 and STK11. When the tumor reaches 180mm 3 When the average size of the tumor was 100 mg / kg, the mice were randomly divided into treatment groups (n=10 / group) and administered the indicated levels of vehicle, 100 mg / kg Formula (10b), 100 mg / kg Sotolacib, or a combination of 100 mg / kg Formula (10b) and 100 mg / kg Sotolacib by oral gavage once a day from day 1 to day 21. The tumor volume and body weight of the mice were measured twice a week until day 21.

[0423] like Figure 3A As shown in Table 5, the combination of Formula (10b) and Sotolacib showed significantly greater anti-tumor activity than either test substance treatment as a monotherapy. Following oral administration of the combination of Formula (10b) (100 mg / kg) and Sotolacib (100 mg / kg) once daily, a statistically significant reduction in tumor volume ( 0.0447 W / m) was observed in the combination group compared to the two monotherapy groups. Figure 3Aand Table 5), and tumor growth inhibition increased significantly to 96% on day 21 (Table 5). The results also showed that both Formula (10b) and Sotolacib had monotherapy antitumor activity at the tested dose levels. After oral administration of Formula (10b) (100 mg / kg) or Sotolacib (100 mg / kg) once daily, a statistically significant decrease in tumor volume was observed in both treatment groups compared to the vehicle group ( Figure 3A and Table 5), and the tumor growth inhibition on day 21 in the formula (10b) and sotolacib monotherapy groups was 58% and 72%, respectively (Table 5).

[0424] Table 5: Summary of the antitumor activity of formula (10b) and sotolacib as monotherapy and in combination in BALB / c nude mice bearing tumors derived from the NCI-H2122 subcutaneous cell line.

[0425]

[0426] Abbreviations: ANOVA = analysis of variance; BALB = Bagg albino; ​​n = number; SEM = standard error of the mean; TGI = tumor growth inhibition.

[0427] Note: Oral dosing was performed daily from day 1 to day 21, and the last tumor measurement was performed on day 21. For statistical analysis: Two-way repeated measures ANOVA of group means followed by post hoc Tukey's multiple comparison test was performed from day 3 to day 21 for vehicle group comparison*, and two-way repeated measures ANOVA of monotherapy and combination group means was performed from day 3 to day 21 for combination group comparison**.

[0428] like Figure 3B As shown in the mouse model, Sotolacib monotherapy or combination treatment had no effect on body weight, and these treatments were well tolerated. Unexpectedly, several mice in the Formula (10b) monotherapy group had a weight loss of more than 10%, so from day 11 to day 21, the animals in this group were placed on a diet gel ( Figure 3B The reason for this weight loss is unknown and has not been observed in previous studies at this dose level (100 mg / kg). Importantly, Formula (10b) was administered at the same dose level (100 mg / kg) in the combination group of Formula (10b) and Sotolacib, and the combination was well tolerated ( Figure 3B ).

[0429] D. Conclusion

[0430] The results of these two in vivo studies showed that the combination of formula (10b) and sotolacib was well tolerated and showed significant anti-tumor benefit in a mutant KRAS G12C NSCLC NCI-H358 cell line-derived xenograft model and a mutant KRAS G12C and KEAP1A203fs NSCLC NCI-H2122 cell line-derived xenograft model. These in vivo studies support the clinical evaluation of the combination and show that SHP2 inhibition can enhance the response of KRASG12C inhibitors and increase the number of patients who can benefit from the combination of formula (10b) and sotolacib as a potential treatment, these patients have mutant KRASG12C, and with or without co-mutations, such as KEAP1 or STK11.

[0431] Example 3: Phase 1A / 1B study of the combination of the SHP2 inhibitor formula (10b) and the KRAS G12C inhibitor sotolacib in patients with solid tumors harboring KRAS G12C mutations

[0432]

[0433]

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442] Figure 5 A Phase 1A / 1B study of the combination of Formula (10b) and sotolacib in patients with solid tumors harboring a KRAS G12C mutation is shown. The study design includes Phase 1a dose escalation and Phase 1b dose expansion / optimization.

[0443]

[0444]

[0445]

[0446] Abbreviations and footnotes for Table 6:

[0447] Abbreviations: AE = adverse event; aPTT = activated partial thromboplastin time; ctDNA = circulating tumor deoxyribonucleic acid; CR = complete response; CT = computed tomography; ECG = electrocardiogram; ECHo = echocardiogram; ECOG = Eastern Cooperative Oncology Group; EOS = end of study; EOT = end of treatment; ET = early termination; DLT = dose-limiting toxicity; FDA = Food and Drug Administration; FSH = follicle-stimulating hormone; FU = follow-up; HBV = hepatitis B virus; HCV = hepatitis C virus; HIV = human immunodeficiency virus; ICF = informed consent; IN R = international normalized ratio; IV = intravenous; KRAS = Kirsten rat sarcoma viral oncogene homolog; LVEF = left ventricular ejection fraction; MRI = magnetic resonance imaging; MUGA = multiple gated acquisition scan; NA = not applicable; NSCLC = non-small cell lung cancer; PD = progressive disease; PET = positron emission tomography; PK = pharmacokinetics; PR = partial response; PT = prothrombin time; QD = once daily; QTcF = QT using Fridericia correction formula; RECIST = response evaluation criteria in solid tumors; RP1bD = recommended phase 1b dose

[0448] A. Due to the coronavirus disease 2019 (COVID-19) pandemic, telephone or video contact visits may be required for safety monitoring, changes in visit schedules, or other appropriate changes in clinical trial conduct. Allowable study days are expected to be specified as best practices; however, adjustments may be made as needed. If necessary, additional changes to the timeline of events will be made and documented in accordance with current FDA guidance (e.g., FDA Guidance for Clinical Trials of Medical Products During the COVID-19 Public Health Emergency).

[0449] B. In each 28-day cycle, the study treatment will be taken orally every 24 hours (± 3 hours). Compound (10b) should be taken before Sotolacib. The day before and on the day of the study visit, the two study treatments should be taken within a 5-minute window unless one of the study treatments is shelved. The study treatment should be taken with about 240mL (8oz) of water after fasting overnight (minimum 8hr), followed by fasting for 2hr after taking the dose. It was opened 1 hour before administration, and water was allowed to be drunk freely. Compound (10b) will be taken QD, combined with Sotolacib QD. Two doses of compound (10b) selected from the dose escalation of Phase 1a will be used for compound (10b) in Phase 1b dose expansion / optimization. The dose can be adjusted based on the SRC decision after reviewing all the data.

[0450] C. Documentation of KRAS G12C mutation confirmed from local or central laboratory testing of tumor or liquid biopsy specimens collected within 2 years prior to screening. Note: KRAS G12C status was determined by a central laboratory, as local laboratory confirmation was not required prior to enrollment.

[0451] D. For Phase 1a: Patients with locally advanced or metastatic solid tumors and KRAS-G12C mutations. For Phase 1b: Patients with locally advanced or metastatic NSCLC and KRAS-G12C mutations who have not been treated with KRAS-G12Ci.

[0452] E. Re-staging scans (CT, MRI, or PET-CT) should be performed at Screening, C3D1 (±7d), and then every 8 weeks (±7d) thereafter until PD. Scans will be performed at the EOT follow-up visit unless performed within 28d of the EOT follow-up visit. Confirmatory scans will also be obtained at least 4 weeks after the initial documented objective response (i.e., PR or CR according to RECIST v1.1). Scans obtained The type was determined by the investigator at their discretion as to whether it was appropriate for the disease. However, the same approach should be used during the duration of the study. All scans will be read locally; in Phase 1b dose expansion / optimization, scans will also be read by blinded independent central review. Patients who discontinue study drug for reasons other than radiological PD will have a radiological assessment during the LTFU period per standard of care imaging, with data reported as they become available until radiological PD is observed (unless consent is withdrawn).

[0453] F. Blood samples will be used to assess relevant tumor markers as needed.

[0454] G. Confirm eligibility prior to administration.

[0455] H. Include year of birth, gender, height, race, ethnicity as permitted by regulations in each region.

[0456] I. Include relevant medical history, current medical conditions, oncology history (e.g., diagnosis, extent and stage of cancer, previous anticancer therapy), radiation history.

[0457] J. Complete physical examination at Screening and EOT Follow-up Visits or ET (if applicable); focused examination at other visits (e.g., based on symptoms indicated at the investigator's discretion). Both the complete and focused examinations include weight.

[0458] K. Vital signs should be taken before any blood draw.

[0459] L. SAEs should be collected from the time of informed consent. Deaths attributed solely to disease progression should not be reported as SAEs, but all accompanying deaths must be recorded on the relevant CRF page.

[0460] M. Predose assessments of hematology, chemistry, coagulation, pregnancy, and urinalysis may be performed up to 2 days prior to the scheduled visit. If these screening assessments are performed within 48 hours prior to Cycle 1, Day 1, these results may be used as a baseline (predose assessment) without the need for a predose assessment within 2 hours of dosing, as described in footnotes W, Y.

[0461] N. Women who have not been surgically sterilized to confirm menopausal status.

[0462] O. Females who are not surgically sterilized or serum tested postmenopausal at all indicated time points. Patients must have a negative test within 48 hrs of D1 of each cycle.

[0463] P. Complete blood count with differential.

[0464] Q. For a list of parameters in the clinical chemistry panel, see Section 8.2.2. [NOTE: Patients should fast for all screening labs, all predose labs, and all labs until 2 hours post-dose]

[0465] R. Includes ctDNA. Samples should be collected after at least 7 consecutive days of compound (10b) administration and may be excluded from sample collection if a visit cannot be scheduled within the allowed visit window.

[0466] S. The medical history and physical examination should be confirmed and updated with any findings that may have occurred prior to the first dose of C1D1.

[0467] T. A fresh tumor biopsy should be collected from all patients enrolled in the study, if at all feasible. If a fresh tumor biopsy was not completed during molecular prescreening and is not feasible during screening, an archived tumor biopsy with sufficient tumor for central testing for KRAS mutations, preferably collected within 6 months but no longer than 2 years of enrollment, may be used. Patients may be enrolled even when a tumor biopsy is judged unavailable in consultation with the Medical Monitor in conjunction with the Investigator. Enrollment of patients without tumor biopsy collection will be handled on a case-by-case basis by the Medical Monitor. Please refer to the Laboratory Specimen Manual for details regarding the collection and handling of tumor tissue samples.

[0468] U. If the start of the subsequent cycle is delayed due to the interruption of the simultaneous administration of two study drugs, the weekly assessment should be recorded as an unscheduled visit for the just completed 28-day cycle until the administration of at least one study treatment is resumed. The first day of the next cycle will be the day of restarting the study treatment. If compound (10b) or sotolacib is interrupted for 15 consecutive days, the patient should permanently interrupt both study drugs.

[0469] V. The Cycle 1 Day 1 visit should be conducted on Monday, Tuesday, Wednesday, or Thursday.

[0470] W. Collection pre-dose (within 2 hr prior to study treatment) and 4 hr (± 15 min) after administration of study treatment.

[0471] X. Unless completed in the previous 7 days.

[0472] Y. Pre-dose data were collected within 2 hours before study treatment administration on the day of the visit.

[0473] Z. Collection pre-dose (i.e., approximately 24 hr [± 2 hr] after study treatment administration on the previous day but before study treatment administration on the day of visit)

[0474] AA. For compound (10b) and sotolacib PK: Collect pre-dose blood samples (within 2 hr before study treatment administration) and 0.5 (±5 min), 1 (±10 min), 2 (±15 min), 4 (±15 min) and 6 hr (±15 min) after administration of study treatment. Steady-state (C2D1) PK samples should be collected after at least 7 consecutive days of each study treatment administration. If a visit cannot be scheduled within the allowed visit window, all samples can be excluded from sample collection.

[0475] BB. If a patient does not continue in a subsequent cycle (ie, does not proceed on study), the patient should still undergo all Day 1 visit procedures for the subsequent cycle, except for the 2-hour PK and pharmacodynamic samples.

[0476] CC. Window (±3d) allowed for each MUGA / ECHO assessment starting at cycle 2.

[0477] DD. Post-dose collection between 2 and 6 hours after study treatment administration on Cycle 2, Day 1 (±7 d).

[0478] EE. Pre-dose samples were collected 2 hr before and 2 hr (±15 min) after study treatment administration on the day of the visit. Samples should be collected after at least 7 consecutive days of study treatment administration. If a visit cannot be scheduled within the allowed visit window, all samples may be excluded from collection.

[0479] FF. 12-lead ECG will be performed at Screening and on C1D1, C2D1, C3D1, and every 2 weeks thereafter, repeated three times.

[0480] GG. Unless completed within 4 weeks of the EOT follow-up visit.

[0481] HH. A MUGA scan or ECHO will be performed at the EOT follow-up visit and ET visit only if LVEF assessment has not been performed within the last 14 days.

[0482] II. Patients who interrupt treatment because of radioactive PD, and patients who interrupt treatment for reasons other than PD and subsequently have radioactive PD will track survival status and new anticancer therapy information approximately every 3 months (by phone) until EOS. This survival follow-up is also applicable to patients who interrupt study treatment for reasons other than radioactive PD but withdraw consent for further tumor assessment. Survival visits will continue until all patients have interrupted study treatment, died, withdrew consent or lost to follow-up, or until EOS, whichever occurs first. Long-term follow-up will also include collecting disease response and secondary deterioration. If arranged as part of a routine visit, a clinical visit may also replace a long-term visit telephone contact.

[0483] JJ.C1D22 will be a telephone visit during which daily self-administration of study treatment will be reviewed from the patient diary and information on AEs and concomitant medications will be recorded in source documents and transcribed to the CRF.

[0484] KK. Molecular prescreening for KRAS mutations by a CLIA-certified laboratory using appropriate clinically validated and / or FDA-approved tests, if applicable, was performed only if there was no documentation of a KRAS mutation within 2 years prior to screening.

[0485] LL. Concomitant medications include all prescription and over-the-counter medications, vaccines, or dietary supplements (vitamins, minerals, herbs)

[0486] MM. For details of the pharmacodynamic evaluations, see Section 8.5 of the clinical protocol.

[0487] NN. To be eligible for the DLT evaluation period, patients need to take 21 doses within the 28-day evaluation period. For additional details, see Sections 4.1.2 and 9.1.1 of the clinical protocol.

[0488] Table 1: Planned Phase 1 Dose Escalation

[0489]

[0490] Abbreviations: PO = oral; QD = once daily; RP1bD = proposed phase 1b dose; SRC = safety review committee

[0491] a If deemed necessary by the SRC, additional patients may be enrolled in the current cohort to evaluate safety, tolerability, and RP1bD.

[0492] b Dose escalation, other dose levels, alternative dose levels, and / or alternative dose schedules may be enrolled after the SRC reviews all data and after the SRC approves or makes a recommendation regarding the next dose cohort.

[0493] A DLT was defined as an AE or abnormal laboratory value that met any of the following criteria, excluding toxicities clearly related to disease progression or intercurrent illness, and occurred during the first cycle (28 days) of the study:

[0494] Grade 3 laboratory abnormalities lasting more than 7 days or Grade 4 laboratory abnormalities of aspartate aminotransferase / alanine aminotransferase (AST / ALT) and / or bilirubin of any duration, except for the following:

[0495] o For patients with grade 1 AST / ALT (>ULN to 2.5×ULN) at baseline, AST / ALT values ​​>7.5×ULN will be considered a DLT

[0496] Grade 3 or higher non-hematologic toxicity, excluding:

[0497] ο Grade 3 nausea, vomiting, or diarrhea that persists for less than 72 hours despite adequate supportive care

[0498] o Grade 3 fatigue lasting less than one week

[0499] o Grade 3 electrolyte abnormalities that persist for less than 72 hours, are clinically uncomplicated, and resolve spontaneously or with routine medical intervention

[0500] o Grade 3 amylase or lipase lasting less than 72 hours and not associated with manifestations of pancreatitis

[0501] Meet Hy's rule criteria (ie, ≥3×ULN ALT and / or AST, ≥2×ULN total bilirubin, and alkaline phosphatase <2×ULN), unless the patient has congenital hyperbilirubinemia.

[0502] Grade 3 neutropenia for >7 days.

[0503] Grade 4 neutropenia.

[0504] Febrile neutropenia [ANC <1000 / mm 3 , with a single oral temperature >38.3°C (101°F) or with two consecutive oral temperatures ≥38°C (100.4°F) measured at least 1 hour apart].

[0505] Grade 3 thrombocytopenia with grade 2 or higher bleeding.

[0506] Grade 4 thrombocytopenia or thrombocytopenia requiring platelet transfusion.

[0507] Grade 4 life-threatening anemia.

[0508] · Grade 5 toxicity.

[0509] Any other clinically significant or persistent toxicity of grade ≥ 3 that could be considered a DLT after SRC review.

[0510] Adverse events (AEs)

[0511] An AE is an adverse medical condition or worsening of a pre-existing medical condition in a patient participating in a clinical study, whether or not it is considered to be causally related to the study drug. Therefore, an AE may be any unfavorable and unexpected sign (including abnormal laboratory findings), symptom, or disease (new or worsening). In clinical studies, AEs may include adverse medical conditions that occur at any time, including baseline or washout periods, even when study treatment is not being administered.

[0512] All AEs that occur after the patient receives the first dose of study drug or during participation in the study must be recorded on the appropriate form provided, regardless of whether they are related to the study drug. Evaluate the severity, intensity, and causal relationship of each AE to each study drug or other factors. SAEs should be recorded and reported from the time of informed consent.

[0513] At each visit, patients should be asked in an open-ended manner about their AEs and about the status of any previously reported AEs.

[0514] When possible, the specific disease or condition rather than the individual associated sign or symptom should be identified and recorded on the form by the investigator. If a sign or symptom is observed or reported that is not considered by the investigator to be a component of a specific disease or condition, it should be recorded on the form as a separate AE.

[0515] NOTE: Unexpected worsening or aggravation of the condition / indication under study should be reported as an AE. However, expected day-to-day fluctuations or expected progression of the disease under study (based on the investigator's clinical judgment) are not considered AEs. Disease progression should be recorded on the relevant CRF page. Deaths attributed solely to disease progression should not be reported as SAEs, but all accompanying deaths must be recorded in the CRF.

[0516] Abnormal laboratory results (e.g., clinical chemistry, hematology, and urinalysis), specific studies (e.g., scans, MUGA / ECHO), or other abnormal assessments (e.g., physical examination results, ECG, and vital signs) that are judged by the investigator to be clinically significant, i.e., require further investigation and / or treatment, will be recorded as AEs. Clinically significant abnormal laboratory results, specific studies or other abnormal assessments detected during the study or present at baseline and significantly worsened after exposure to study treatment are reported as AEs. However, clinically significant abnormal laboratory results, specific studies or other abnormal assessments related to the disease being studied should not be reported as AEs unless judged by the investigator to be more severe than expected for the patient's condition or to be present at the start of the study but not to worsen.

[0517] Medical or surgical procedures (e.g., endoscopy, appendectomy) are not AEs; conditions requiring surgery if new or worsened from baseline are AEs. Elective procedures or pre-planned interventions for pre-existing conditions that would not have worsened from the start of the study are also not AEs; pre-existing conditions should be documented as medical history. Hospitalizations for social reasons and / or convenience without adverse medical events are not AEs.

[0518] The investigator is responsible for reviewing all laboratory test results for all patients, including laboratory evaluations performed at outside laboratories, and determining whether they constitute an AE. The investigator will evaluate the outside laboratory test results and determine the clinical significance of any considered AEs. In these cases, the laboratory report should be signed and dated by the investigator.

[0519] Serious Adverse Events (SAEs)

[0520] SAEs are any of the following AEs occurring at any dose regardless of causality:

[0521] Causes death, or death is a consequence of the AE rather than the AE itself, unless the cause of death is unknown, in which case the death should be recorded as an AE.

[0522] Life-threatening

[0523] o Life-threatening means that, in the opinion of the investigator or study sponsor, the patient is at immediate risk of death as a result of the reaction (ie, it does not include reactions that have occurred in a more severe form and are hypothetically likely to cause death).

[0524] · Requires hospitalization of the patient or extension of existing hospitalization.

[0525] o In general, hospitalization refers to the retention (usually involving at least an overnight stay) of a patient in a hospital or emergency room for observation and / or treatment that is not appropriate in a physician's office or outpatient setting.

[0526] o A complication that occurred during hospitalization was an AE. An event was serious if the complication prolonged hospitalization or met any other severity criteria.

[0527] o If an ailment or disease existed prior to the patient's enrollment in the trial, a hospital admission and / or surgical procedure was scheduled during the study period, but was planned before the signing of the ICF, it was not considered an AE, provided that it did not worsen in an unexpected manner during the trial (e.g., surgery was performed earlier than planned).

[0528] · Resulting in permanent or significant disability / incapacity.

[0529] οDisability is defined as a substantial impairment of an individual's ability to carry out normal life functions.

[0530] o The definition is not intended to include relatively minor medically significant experiences that may interfere with or prevent daily functioning but do not constitute a substantial impairment, such as uncomplicated headaches, nausea, vomiting, diarrhea, influenza, and accidental trauma (e.g., a sprained ankle).

[0531] Is a congenital anomaly / birth defect.

[0532] It is a major medical event.

[0533] o A significant medical event is an event that may not result in death, be life-threatening, or require hospitalization, but which may be considered a SAE based on appropriate medical judgment, which may endanger the patient and may require medical or surgical intervention to prevent one of the outcomes listed in the definition for an SAE. Examples of such medical events include allergic bronchospasm requiring intensive treatment in the emergency room or at home, blood dyscrasias or seizures that do not result in hospitalization of the patient, or the development of drug dependence or drug abuse.

[0534] A distinction should be made between the terms “serious” and “severe” as they are not synonymous. The term “severe” is often used to describe the intensity (severity) of a specific event (e.g., mild, moderate, or severe myocardial infarction); however, the event itself may be of relatively minor medical significance (e.g., severe headache). This is different from “serious,” which is based on patient / event outcomes or action criteria generally associated with events that threaten patient life or function. A severe AE does not necessarily need to be considered serious. For example, persistent nausea lasting several hours may be considered severe nausea, but it is not an SAE if the event does not meet the severity criteria. On the other hand, a stroke that results in only mild disability may be considered mild but is defined as an SAE based on the severity criteria described above. Severity (not severity) serves as a guide for defining regulatory reporting obligations.

[0535] Intensity of AE usage

[0536] NCI CTCAE v 5.0 rating available at https: / / ctep.cancer.gov / protocolDevelopment / electronic_applications / docs / CTCAE_v5_Quick_Reference_5x7.pdf, the disclosure of which is incorporated herein by reference in its entirety.

[0537] Administration, Dosing Adjustments, Delays, and Interruptions

[0538] The duration of each treatment cycle will be 28 days, unless an alternative dosing schedule is implemented based on SRC review. In the absence of a reason for interruption of dosing, a new cycle will begin immediately after the completion of the previous cycle. Compound (10b) and sotolacib will be self-administered without interruption, unless 1) there is an AE as described below; 2) the SRC recommends an alternative dosing schedule based on observed safety, tolerability and PK / pharmacodynamic data; or 3) the patient is seen in the outpatient clinic to evaluate AEs, PK sample collection or laboratory evaluation. At such visits, patients will take compound (10b) and sotolacib at the same time in the outpatient clinic as directed by the research staff. For any blood draws performed outside the clinic, patients will take compound (10b) and sotolacib as directed by the research staff and record / provide dosing information. Each dose of compound (10b) should be taken orally once every 24 hours (±3 hours) with approximately 240 mL (8 ounces) of water. Patients should take compound (10b) after fasting overnight (minimum 8 hours), followed by a 2-hour fast after taking the dose. Water is allowed ad libitum except for 1 hour before and after dosing. If a dose is missed, the next dose will be increased without taking into account the missed dose. The patient will take the next regularly scheduled dose at the scheduled time.

[0539] Although sotolacib can be taken with or without food, both study treatments should be taken in the fasting state as described for compound (10b). Compound (10b) should be taken before sotolacib. Both study treatments should be taken within a 5-minute window on the day before and on the day of the study visit.

[0540] If Compound (10b) or Sotolacib was permanently discontinued, the patient had to also discontinue the other study treatment; monotherapy with Compound (10b) or Sotolacib was not permitted in this study.

[0541] Reasons for permanent discontinuation of both study treatments (compound (10b) and sotolacib) included, but were not limited to, the following:

[0542] The patient develops a DLT and is unable to continue with a lower dose of study treatment (as discussed elsewhere in this example);

[0543] Permanent discontinuation of either study treatment for reasons outlined elsewhere for compound (10b) or sotolacib;

[0544] Disease progression as assessed by the investigator according to RECIST v.1.1;

[0545] Risk to patients as judged by the investigator and / or sponsor;

[0546] Severe non-compliance with the regimen as judged by the investigator and / or sponsor;

[0547] The patient is pregnant;

[0548] Patient decision making;

[0549] Initiation of non-study anticancer treatment; and

[0550] Failure to restart treatment within 15 days of dose interruption

[0551] Specific reasons for withdrawal from the study will include the following:

[0552] The patient withdraws consent. The patient is free to withdraw from the study at any time without affecting further treatment;

[0553] Lost to follow-up (defined as no contact after 3 documented telephone attempts followed by 1 attempt via confirmation letter);

[0554] Death due to any cause;

[0555] The sponsor decides to terminate the study: and

[0556] Decision of the researcher.

[0557] For each patient, a maximum of two dose reductions of each oral agent (Compound (10b) or Sotolacib) will be allowed. Dose reductions for each oral agent are discussed in detail elsewhere in this example.

[0558] Compound (10b)

[0559] Dose modification decisions for compound (10b) should be based on the CTCAE grading scale v5.0 and according to Table 8. If a drug-related AE is not specified in the table, the dose may be reduced or maintained or permanently interrupted at the discretion of the investigator for the safety of the patient.

[0560] For each patient, a maximum of two dose reductions of Compound (10b) will be allowed. Dose reduction of Compound (10b) means treatment at the next lower previously tested dose level, as summarized in Table 7. If a dose reduction below the lowest previously tested Compound (10b) dose is required, the dose should be selected after discussion between the Investigator and the Medical Monitor.

[0561] Table 8: Dose adjustment of compound (10b) for drug-related adverse events

[0562]

[0563] a These general guidelines constitute guidance to researchers and may be supplemented by discussions with the medical monitor in specific circumstances.

[0564] Other permitted compounds (10b) Dose adjustments

[0565] Treatment interruptions and dose adjustments of compound (10b) other than those mentioned above may be considered following discussion between the Investigator and the Medical Monitor and appropriate documentation of the justification.

[0566] Any changes should be within the following guidelines:

[0567] Further dose reductions of compound (10b) may be performed to maintain clinically significant drug-related AEs ≤ Grade 1. Dose adjustments of more than 1 dose level at a time may be considered when toxicity has resolved, as judged in the best interest of the patient (e.g., severe myelosuppression, lactic acidosis).

[0568] If a Grade 3 AE believed to be related to Compound (10b) is observed, treatment may be delayed for up to 2 weeks to allow for resolution of toxicity, with retreatment at a reduced dose at the discretion of the investigator. If Grade 3 nausea, vomiting, diarrhea, or clinically significant electrolyte abnormalities occur and resolve within 3 days of optimal therapy, treatment may be resumed without dose reduction. If recurrent Grade 3 nausea, vomiting, diarrhea, or clinically significant electrolyte abnormalities occur for >3 days, treatment may be resumed at a reduced dose after resolution of symptoms.

[0569] • If a patient experiences more than one AE, the Compound (10b) dose decision will be based on the most severe AE.

[0570] Patients who require a dose reduction and subsequently experience a recurrent clinically significant drug-related AE may undergo one additional dose reduction. Patients who continue to experience this event after two dose reductions will be withdrawn from the study. The lower dose will be based first on the SRC-cleared lower dose as part of the dose escalation for this study, and if a lower dose is required, based on the SRC-cleared dose in Study NAV-1001.

[0571] Sotolacib

[0572] According to the LUMAKRAS product package insert (LUMAKRAS package insert 2021, the entire contents of which are incorporated herein by reference), two dose reductions of sotolacib are allowed in the case of adverse reactions (reduced to 480 mg / day; reduced to 240 mg / day). If the patient cannot tolerate the minimum dose of 240 mg / day, the drug should be discontinued.

[0573] Dose modifications for sotolacizumab due to adverse reactions are shown in Table 9.

[0574] Table 9: Recommended dose adjustments for sotolacib due to adverse reactions

[0575]

[0576]

[0577] Abbreviations: ALT = alanine aminotransferase, AST = aspartate aminotransferase, CTCAE = Common Terminology Criteria for Adverse Events; ILD = interstitial lung disease, ULN = upper limit of normal

[0578] Note: Rating is based on CTCAE v5.0.

[0579] Source: Product insert LUMAKRAS (LUMAKRAS Drug Insert 2021, the entire contents of which are incorporated by reference)

[0580] Effectiveness Assessment

[0581] A restaging scan (computed tomography [CT], MRI, or positron emission tomography / CT [PET-CT]) will be performed. Confirmatory scans should be obtained at least 4 weeks after the initial documentation of an objective response (i.e., PR or CR according to RECIST v1.1). The type of scan obtained is at the discretion of the investigator as appropriate for the disease. However, the same approach should be used for the duration of the study.

[0582] Patients who discontinue treatment for reasons other than radiological PD will continue to have radiological assessments based on standard of care imaging, with data reported as they become available, or upon initiation of subsequent anticancer therapy, death, or patient withdrawal of consent for radiological assessment, whichever occurs first.

[0583] Tumor response will be assessed using RECIST v1.1 (Eisenhauer Eur J Cancer; 2009; 45(2): 228-247). Tumor response will be assessed locally by the investigator for the study; scans for the Phase 1b portion of the study will also be read by a blinded independent central review (BICR). Progression-free survival will be determined based on the date of the first documented progression or death attributed to any cause.

[0584] Pharmacokinetics (PK) and pharmacodynamics (PD)

[0585] Pharmacokinetics

[0586] Blood samples will be collected from all enrolled patients participating in the study for PK evaluation of compound (10b) and Sotolacib. The time points for blood sample collection for PK of compound (10b.) and Sotolacib are summarized in Table 10. Additional details on sample collection and storage can be found in the laboratory manual.

[0587] Table 10: Compound (10b) and Sotolacib PK sample collection schedule

[0588]

[0589] A. Collection was pre-dose (ie, approximately 24 hr [± 2 hr] after administration of study treatment on the previous day, but prior to study treatment administration on the day of the visit).

[0590] On the day of PK sampling, patients should take study medication as directed by study staff in the clinic. Patients who forget and take their medication at home will be excluded from the PK analysis on that day; blood samples should not be collected from them. Complete dosing information, including the date and time of the actual blood draw and the time of the last study treatment dose before sampling, should be obtained on all sampling days and recorded on the appropriate blood collection CRF. If any of the scheduled sampling times are missed or samples are not drawn according to the schedule, the actual collection date and time will be recorded and the remaining samples will be collected as scheduled as possible.

[0591] Steady-state (C2D1 and beyond) PK samples should be collected after at least 7 consecutive days of each study treatment administration. If a study visit cannot be rescheduled within an acceptable window, PK samples should not be drawn.

[0592] If vomiting occurs within 4 hours of compound (10b) or sotolacib administration on the day of PK blood sampling, the time of vomiting should be recorded (using a 24-hour clock). No other compound (10b) or sotolacib should be administered to replace the vomited substance. The occurrence and frequency of any vomiting during the treatment cycle must be recorded in the AE CRF. PK collections should continue as planned in all cohorts.

[0593] Pharmacodynamics

[0594] Blood and tumor samples will be used to evaluate the pharmacodynamic effects of the combination of compound (10b) and sotolacib and potential mechanisms of anti-tumor response and resistance, including but not limited to the evaluations provided in Table 11. These samples will also be used to develop biomarkers that can be used to predict response or resistance to the combination of compound (10b) and sotolacib.

[0595] If at all feasible, a fresh tumor biopsy should be collected from all patients enrolled in the study. If a fresh tumor biopsy was not completed during molecular prescreening and is not feasible during screening, an archived tumor biopsy that was preferably collected within 6 months but no longer than 2 years of enrollment and has sufficient tumor for central testing of KRAS G12C mutations may be used. Fine needle aspirates or other cytology samples are not acceptable.

[0596] Patients may be enrolled even when a tumor biopsy is not available in the judgment of the Medical Monitor in consultation with the Investigator. Enrollment of patients without tumor biopsy collection will be handled on a case-by-case basis by the Medical Monitor.

[0597] The samples obtained will be used in various biomarker studies, including:

[0598] Gene expression within biopsied tumor tissue and circulating in the blood to identify genes or gene markers whose expression is modulated by compound (10b) and which can serve as pharmacodynamic biomarkers.

[0599] Molecular analysis (e.g., NGS, PCR, etc.) of tumor biopsies and ctDNA (liquid biopsies) to evaluate potential mechanisms of drug resistance.

[0600] pSHP2 and pERK were assessed by immunohistochemistry in tumor tissues as indicators of MAPK pathway flux.

[0601] Tumor microenvironment (TME) analysis in tumor biopsies by IHC or multiplex immunofluorescence as an indicator of tumor immune cell infiltration and activation.

[0602] Table 11: Pharmacodynamic evaluation

[0603]

[0604] A. Samples should be collected after administration of compound (10b) for at least 7 consecutive days.

[0605] BOIN Design

[0606] A BOIN design with a target toxicity rate of φ = 0.25 will be used to find the MTD of the combination of compound (10b) and sotolacib. Figure 6 As shown in , the BOIN design uses the following rules, optimized to minimize the probability of incorrect dose assignment, to guide dose escalation / de-escalation:

[0607] If the observed DLT rate at the current dose is ≤ 0.197, the dose will be escalated to the next higher dose level;

[0608] If it is ≥ 0.298, the dose is decreased to the next lower dose level;

[0609] .Otherwise, remain at current dose.

[0610] For the purpose of overdose control, if Pr(p j >0.25|data)>0.95, then dose j and higher levels will be eliminated from further examination, where p j is the true DLT rate at dose level j, j = 1, …, 3. This posterior probability is based on the β-binomial model y j |p j ~ Binomial (p j ) evaluation, where p j ~ uniform (0, 1), where y j is the number of DLTs experienced by patients at dose level j. The trial was terminated for safety when the lowest dose was eliminated. The probability cutoff of 0.95 was chosen to conform to the convention that when the target DLT rate Φ ≤ 1 / 6, the dose at which 3 / 5 patients experienced DLT was eliminated.

[0611] The steps to implement the BOIN design are described below:

[0612] 1. To assign doses to the next cohort of patients (phase to be completed by the SRC), dose escalation / decrease will be performed according to the rules shown in Table 12 and based on a target toxicity rate of 25%. When using Table 12, please note the following:

[0613] a. "Eliminate" means eliminating current and higher doses from the trial to prevent treatment of any future patients at these doses because of excessive toxicity.

[0614] b. When a dose is eliminated, the dose is automatically reduced to the next lower level. When the lowest dose is eliminated, the trial will be terminated for safety. In this case, no dose should be selected as the MTD.

[0615] c. If none of these actions (ie, ramp up, ramp down, or elimination) are triggered, new patients will be treated at the current dose.

[0616] d. If the current dose is the lowest dose and the rule indicates dose reduction, new patients will be treated at the lowest dose unless the number of DLTs reaches the elimination boundary, at which point the trial will be terminated for safety.

[0617] e. If the current dose is the highest dose and the rule indicates dose escalation, the new patient will be treated at the highest dose.

[0618] f. In the event that operational / practical circumstances result in overenrollment (i.e., >7 DLT-evaluable patients per current cohort size), the next dose level decision will be based on the actual number of DLT-evaluable patients in the cohort.

[0619] 2. Dose escalation will start with 5-7 DLT evaluable patients enrolled in compound (10b) 250 mg plus sototraxib 960 mg (i.e., the lowest dose level). According to Table 12, for illustrative purposes, if a DLT is observed in any of the 6 evaluable patients enrolled, then escalate to the next dose level. If DLT is observed in ≥2 / 6 evaluable patients enrolled, then decrement to a lower dose.

[0620] 3. If escalating to a higher dose level, follow the same approach as in 2.

[0621] 4. The maximum sample size for dose escalation is 21 DLT-evaluable patients and 10 DLT-evaluable patients in any dose level cohort.

[0622] Table 12: Dose escalation decision rules

[0623]

[0624] Abbreviation: DLT = dose-limiting toxicity

[0625] a When none of these actions (ie, escalation, decrementation, or elimination) are triggered, stay at the current dose for treatment of the next cohort of patients. Note that the "number of DLTs" is the number of patients with at least 1 DLT.

[0626] After the trial is completed, the MTD will be selected based on ordinal regression as specified in Liu and Yuan (2015). This calculation is implemented by the "estimate MTD" of the BOIN design desktop program. In particular, the dose whose isotonic estimate of the DLT rate is closest to the target DLT rate is selected as the MTD. If there is a connection, a higher dose level is selected when the isotonic estimate is lower than the target DLT rate, and a lower dose level is selected when the isotonic estimate is higher than or equal to the target DLT rate. The RP1bD for cohort expansion will be selected based on toxicity (i.e., MTD) and other clinical considerations (e.g., PK / pharmacodynamics).

[0627] Table 13 shows the operational characterization of the trial design based on 1000 simulated trials using the BOIN design desktop program (MD Anderson Cancer Center, 2021). The operational characterization shows that the design selects the true MTD with a high probability (if any) and allocates more patients to the dose level with the DLT rate closest to the target 0.3.

[0628] Table 13: Operating characteristics of BOIN dose results from simulations

[0629]

[0630]

[0631] Abbreviations: BOIN = Bayesian Optimal Interval Design; DL = highest dose cleared by SRC in ongoing compound (10b) monotherapy trial (NCT04528836); DLT = dose-limiting toxicity

[0632] a Dose escalation, alternative dose levels, and / or alternative dose schedules will be performed after the SRC reviews all safety data and after the SRC approves or makes a recommendation regarding the next dose cohort.

[0633] Note: “% Early Termination” refers to early terminations due to excessive DLTs.

[0634] Effectiveness

[0635] Patient Evaluation & Statistics

[0636] Patients with solid tumors will undergo disease assessments at Screening and then every 8 weeks thereafter unless more frequent assessments are warranted clinically or per RECIST v1.1 criteria outlined herein.

[0637] Descriptive statistics will be provided for continuous variables, and frequency tables will be used to summarize categorical variables. Best overall response and proportion of patients experiencing CR, PR, SD, and PD among patients with measurable disease will be shown, along with the corresponding 95% exact CIs.

[0638] Time-to-event variables (eg, DOR, PFS, and OS) will be analyzed by the Kaplan-Meier method. The median, as well as the 25%, 75%, minimum, and maximum, time-to-event values ​​will be estimated, and Kaplan-Meier curves will be plotted.

[0639] Anti-tumor effect - solid tumors

[0640] Tumor response is the primary efficacy endpoint in Phase 1b dose expansion / optimization; Patients will be evaluated with standard criteria (note: all patients must have measurable disease when enrolled in the study, and will therefore be evaluated according to RECIST v1.1 criteria). For the purpose of this study, tumor response should be evaluated every 8 weeks (± 7 days) (i.e., the 1st day of each odd-numbered cycle), thereafter until PD, while the patient is undergoing treatment. Disease assessments for patients interrupted for reasons other than PD are summarized in Section 7.6 of the clinical protocol. In addition to baseline scans, confirmation scans will also be obtained at least 4 weeks after the first recording of objective response (i.e., PR or CR).

[0641] In this study, response and progression will be evaluated using the new international standard RECIST v1.1 proposed by the RECIST committee (Eisenhauer et al., Eur J Cancer; 2009; 45(2): 228-247). Only the change in the largest diameter (a one-dimensional measurement) of the tumor lesion is used in the RECIST criteria.

[0642] Disease parameters

[0643] Measurable disease. Measurable lesions were defined as those that could be accurately measured >10 mm in at least one dimension (the longest diameter [LD] to be recorded) by scanning (CT, MRI, or PET-CT; scan slice thickness should not exceed 5 mm). Millimeters (or decimal fractions of centimeters).

[0644] Lesions cannot be measured. All other lesions (or disease sites), including small lesions (LD <10 mm or pathological lymph nodes ≥10 mm to <15 mm), were considered non-measurable disease. Bone lesions, leptomeningeal disease, ascites, pleural / pericardial effusions, cutaneous / pulmonary lymphangitis, mastitis, abdominal masses (without subsequent CT, MRI, or PET-CT), and cystic lesions were all considered true non-measurable disease.

[0645] Target lesion. All measurable lesions representing all involved organs should be identified as target lesions at baseline, with a maximum of 2 lesions per organ and a total of 5 lesions, and recorded and measured. Target lesions should be selected based on their size (lesions with LD) and suitability for accurate repeated measurements (by imaging counts or clinically). The sum of the LD of all target lesions will be calculated and reported as the baseline sum LD. The baseline sum LD will be used as a reference to characterize the objective tumor response.

[0646] Non-target lesions. All other lesions (disease sites) (including any measurable lesions in excess of and above the 5 target lesions) should be identified as non-target lesions at baseline and should also be recorded. These lesions do not need to be measured, but the presence or absence of each lesion should be noted throughout follow-up.

[0647] Methods used to assess measurable disease

[0648] All measurements should be made using a ruler or calipers and recorded in metric notation. All baseline evaluations should be performed as close to the start of treatment as possible and never more than 4 weeks before the start of treatment.

[0649] Each identified and reported lesion should be characterized using the same assessment method and the same technology at baseline and during follow-up. When both methods have been used to assess the antitumor effect of treatment, imaging-based evaluation is superior to evaluation by clinical examination.

[0650] CT, MRI, or PET-CT scanning should be used to measure selected lesions for response assessment : These techniques should be performed with serial sections ≤5 mm thick. Each identified and reported lesion should be characterized using the same assessment method and the same technique at baseline and during follow-up.

[0651] Tumor markers. Tumor markers alone cannot be used to assess response. If a marker is initially above the upper limit of normal, it must be normalized so that the patient is considered to have a complete clinical response. Specific additional criteria for standardized use of prostate-specific antigen and CA-125 responses to support clinical trials are being developed.

[0652] Cytology, Histology: These counts can be used to differentiate between PR and CR in rare cases (eg, residual lesions in tumor types such as germ cell tumors where residual benign tumor is known to remain).

[0653] When measurable tumor meets the criteria for response or stable disease, cytologic confirmation of the neoplastic origin of any effusion that appears or worsens during treatment is mandatory to distinguish between response or stable disease (effusions can be a side effect of treatment) and PD.

[0654] Reaction criteria

[0655] Evaluation of target lesions

[0656] Complete Response (CR): All target lesions disappeared.

[0657] Partial Response (PR): At least 30% reduction in the sum of LD of all target lesions, using the baseline sum LD as reference.

[0658] Progressive disease (PD): The sum of the target lesion LDs increased by at least 20%, taking as reference the minimum sum LD recorded since the start of treatment or the appearance of one or more new lesions.

[0659] Stable disease (SD): If the disease was neither sufficiently reduced to meet the criteria of PR nor sufficiently increased to meet the criteria of PD, the smallest summed LD from the start of treatment was used as the reference.

[0660] Evaluation of non-target lesions

[0661] Complete Response (CR): All non-target lesions disappeared and tumor marker levels normalized.

[0662] NOTE: If tumor markers are initially above the upper limit of normal, they must be normalized in order for the patient to be considered in complete clinical response.

[0663] Stable disease (SD): Persistence of one or more non-target lesions and / or maintenance of tumor marker levels above normal limits.

[0664] Progressive disease (PD): Appearance of one or more new lesions and / or unequivocal progression of existing non-target lesions.

[0665] Although unequivocal progression of only “non-target” lesions is an exception, in such cases the opinion of the attending physician should prevail and progression status should be confirmed later by the review team (or the principal investigator).

[0666] Best overall response evaluation

[0667] Best overall response is the best response recorded from the start of treatment until disease progression / relapse (for PD, the smallest measurement recorded from the start of treatment is used as reference). The patient's best response assignment will depend on the achievement of measurement and confirmation criteria.

[0668] Table 14: Evaluation of Best Overall Response

[0669]

[0670] Reaction duration

[0671] Duration of overall response: DOR was measured from the time when CR or PR (whichever was recorded first) was met until the first date of objectively documented relapse or PD (for PD, the smallest measurement recorded from the start of treatment was used as reference).

[0672] Duration of overall CR was measured from the time when CR criteria were first met until the first date of objectively documented recurrent disease.

[0673] Duration of disease stability: Stable disease was measured from the start of treatment until progression criteria were met, using the smallest measurement recorded from the start of treatment as the reference.

[0674] Progression-free survival

[0675] Progression-free survival (PFS) will be defined as the treatment start date to the event date, defined as the first documented progression or death from any cause. If the patient does not experience an event, PFS will be censored according to the details described in the SAP. Kaplan-Meier analysis of PFS will be provided.

[0676] Although the foregoing disclosure has been described in some detail by way of illustration and example for the sake of clarity of understanding, it will be appreciated by those skilled in the art that certain changes and modifications may be implemented within the scope of the appended claims. In addition, each reference provided herein is incorporated by reference in its entirety to the same extent as each reference is incorporated by reference individually. If there is a conflict between the present application and the references provided herein, the present application shall prevail.

Claims

1. A method of treating cancer in a subject, comprising administering to the subject: a) a therapeutically effective amount of a compound represented by formula (I): or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformer, tautomer, or combination thereof; and b) a therapeutically effective amount of sotolacib (AMG 510).

2. The method according to claim 1, wherein the compound of formula (I) is represented by formula (10b): The name is 6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]dec-8-yl)-3-(R a )-(2,3-dichlorophenyl)-2,5-dimethylpyrimidin-4(3H)-one.

3. The method of claim 1 or claim 2, wherein the cancer is characterized by a KRAS mutation.

4. The method of claim 3, wherein the cancer is characterized by a KRAS G12C mutation.

5. The method of any one of claims 1 to 4, wherein the cancer comprises a solid tumor.

6. The method of any one of claims 1 to 5, wherein the cancer is lung cancer, colorectal cancer, pancreatic cancer, urothelial carcinoma, gastric cancer, mesothelioma, or a combination thereof.

7. The method of claim 6, wherein the cancer is non-small cell lung cancer (NSCLC).

8. The method of any one of claims 1 to 7, wherein the cancer is a KRAS G12C-positive cancer that is resistant to a KRAS G12C inhibitor.

9. The method of any one of claims 1 to 8, wherein the cancer is a KRAS G12C-positive cancer characterized by intrinsic and / or acquired resistance to KRAS G12C inhibitors.

10. The method of any one of claims 1 to 9, wherein the cancer is a KRAS G12C-positive cancer that is resistant to sotolacib.

11. The method of any one of claims 1 to 10, wherein the cancer progresses or relapses on or after at least one prior line of systemic therapy comprising platinum-based double chemotherapy and / or anti-PD-1 / PD-L1 therapy, each administered as a monotherapy or both administered as a combination therapy.

12. The method of any one of claims 1 to 11, wherein the subject does not have an activating mutation in BRAF V600X, PTPN11 (SHP2), or KRAS Q61X.

13. The method according to any one of claims 1 to 12, wherein the subject has not been previously treated with a PTPN11 inhibitor.

14. The method according to any one of claims 1 to 12, wherein the subject has been previously treated with a PTPN11 inhibitor other than a compound of formula (I).

15. The method of any one of claims 1 to 12, wherein the subject has been previously treated with a compound of formula (I).

16. The method of any one of claims 1 to 15, wherein the subject has not been previously treated with a KRAS G12C inhibitor.

17. The method of any one of claims 1 to 8 and 10 to 15, wherein the subject has been previously treated with a KRASG12C inhibitor.

18. The method of any one of claims 1 to 15, wherein the subject was previously treated with sotolacib.

19. The method according to any one of claims 1 to 18, wherein the subject meets all of the inclusion criteria according to 1) to 11) of Example 3, provided that the subject does not meet any of the exclusion criteria according to 1) to 17) of Example 3.

20. The method according to any one of claims 1 to 19, wherein the subject is a human.

21. The method according to any one of claims 1 to 20, wherein the compound of formula (I) or (10b) and sotolacib are administered concomitantly.

22. The method according to any one of claims 1 to 20, wherein the compound of formula (I) or (10b) and sotolacib are administered sequentially.

23. The method of claim 21, wherein the compound of formula (I) or (10b) is administered prior to administration of sotolacib.

24. The method of claim 21, wherein the compound of formula (I) or (10b) is administered after administration of sotolacib.

25. The method according to any one of claims 1 to 24, wherein the compound of formula (I) or (10b) or sotolacib is administered orally, or sotolacib and the compound of formula (I) or (10b) are each administered orally.

26. The method according to any one of claims 1 to 25, wherein the compound of formula (I) or (10b) is administered orally.

27. The method of any one of claims 1 to 26, wherein sotolacib is administered orally.

28. The method of any one of claims 1 to 27, wherein the compound of formula (I) or (10b) and sotolacib are provided in a combined therapeutically effective amount.

29. The method of any one of claims 1 to 27, wherein the compound of formula (I) or (10b) and sotolacib are provided in synergistically effective amounts.

30. The method of any one of claims 1 to 29, wherein the compound of formula (I) or (10b) and / or sotolacib is used in a dose different from when they are used alone.

31. The method of claim 30, wherein the compound of formula (I) or (10b) is used at a lower dose than when used alone.

32. The method of claim 30, wherein the compound of formula (I) or (10b) is used in a higher dose than when used alone.

33. The method of any one of claims 30 to 32, wherein sotolacib is used at a lower dose than when it is used alone.

34. The method of any one of claims 30 to 32, wherein sotolacib is used at a higher dose than when it is used alone.

35. The method of any one of claims 1 to 34, wherein the treatment comprises one or more treatment cycles; wherein each of the one or more treatment cycles has a duration of about 28 days; and the compound of formula (I) or (10b) and / or sotolacib is administered daily.

36. The method of any one of claims 1 to 35, wherein the administration of the compound of Formula (I) or (10b) and sotolacib comprises one or more dose escalations, dose maintenance, or dose reductions of the compound of Formula (I) or (10b) and / or sotolacib.

37. The method of claim 36, wherein the administration of the compound of Formula (I) or (10b) and sotolacib comprises one or more dose escalations, dose maintenance, or dose reductions of the compound of Formula (I) or (10b).

38. The method of claim 37, wherein the one or more dose escalations, dose holds, or dose reductions of the compound of formula (I) or (10b) are determined by dose limiting toxicity (DLT) assessment.

39. The method of claim 38, wherein said administration of said compound of formula (I) or (10b) comprises dose escalation following a prior treatment cycle when the dose limiting toxicity (DLT) rate is less than about 19.7% as determined by DLT assessment.

40. The method of claim 38, wherein said administration of said compound of formula (I) or (10b) comprises dose reduction following a previous treatment cycle when the rate of dose limiting toxicity is greater than about 29.8% as determined by DLT assessment.

41. The method of claim 38, wherein the administration of the compound of formula (I) or (10b) comprises dose maintenance following a previous treatment cycle when the dose limiting toxicity rate is in the range of about 21.9% to about 29.8% as determined by DLT assessment.

42. The method of any one of claims 35 to 41, wherein the treatment comprises a dose escalation phase, and wherein after the dose escalation phase, the treatment further comprises a dose expansion / optimization phase; and the compound of formula (I) or (10b) is administered at a dosage regimen determined during the dose escalation phase.

43. The method of claim 42, wherein during the dose expansion / optimization phase, the administration of the compound of formula (I) or (10b) comprises one or more dose adjustments.

44. The method of any one of claims 1 to 43, wherein the therapeutically effective amount of sotolacib is a total daily dose of about 120 mg, about 240 mg, about 360 mg, about 480 mg, about 600 mg, about 720 mg, about 840 mg, or about 960 mg.

45. The method of claim 44, wherein the therapeutically effective amount of sotolacib is a total daily dose of about 960 mg.

46. ​​The method of any one of claims 1 to 45, wherein the therapeutically effective amount of the compound of formula (I) or (10b) is from about 100 mg to about 2000 mg, from about 150 mg to about 1000 mg, from about 200 mg to about 1000 mg, from about 250 mg to about 1000 mg, from about 300 mg to about 1000 mg, from about 350 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 450 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 550 mg to about 1000 mg, from about 600 mg to about 1000 mg, from about 650 mg to about 1000 mg, from about 700 mg to about 1000 mg, from about 100 mg to about 700 mg, from about 150 mg to about 700 mg, from about 200 mg to about 700 mg, from about 250 mg to about 1000 mg. g to about 700 mg, from about 300 mg to about 700 mg, from about 3 mg to about 700 mg, from about 400 mg to about 700 mg, from about 450 mg to about 700 mg, from about 500 mg to about 700 mg, from about 550 mg to about 700 mg, from about 100 mg to about 550 mg, from about 150 mg to about 550 mg, from about 200 mg to about 550 mg, from about 250 mg to about 550 mg, from about 300 mg to about 550 mg, from about 350 mg to about 550 mg, from about 400 mg to about 550 mg, from about 450 mg to about 550 mg, from about 100 mg to about 400 mg, from about 150 mg to about 400 mg, from about 200 mg to about 400 mg, from about 250 mg to about 400 mg, or a total daily dose of from about 300 mg to about 400 mg, on a salt-free and water-free basis.

47. The method of claim 46, wherein the therapeutically effective amount is a total daily dose of the compound of formula (I) or (10b) from about 250 mg to about 400 mg, from about 400 mg to about 550 mg, or from about 550 mg to about 700 mg, on a salt-free and water-free basis.

48. The method of claim 47, wherein the therapeutically effective amount is a total daily dose of about 250 mg, about 400 mg, or about 550 mg of the compound of formula (I) or (10b), on a salt-free and water-free basis.

49. The method of any one of claims 46 to 48, wherein the therapeutically effective amount is a total daily dose of about 250 mg of the compound of formula (I) or (10b), calculated on a salt-free and water-free basis.

50. The method of any one of claims 46 to 48, wherein the therapeutically effective amount is a total daily dose of about 400 mg of the compound of formula (I) or (10b) on a salt-free and water-free basis.

51. The method of any one of claims 46 to 48, wherein the therapeutically effective amount is a total daily dose of about 550 mg of the compound of formula (I) or (10b) on a salt-free and water-free basis.

52. The method of any one of claims 1 to 51, wherein the compound of formula (10b) and sotolacib are each administered orally.

53. The method of any one of claims 1 to 52, wherein the compound of formula (10b) is administered once, twice, three times or four times daily.

54. The method of claim 53, wherein the compound of formula (I) or (10b) is administered once daily; and sotolacib is administered once daily.

55. The method of any one of claims 1 to 54, wherein the compound of formula (I) or (10b) is provided in a tablet formulation.

56. The method of any one of claims 1 to 55, wherein the treatment reduces the volume of the cancer or solid tumor by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%.

57. The method of any one of claims 1 to 55, wherein the treatment stabilizes the cancer or solid tumor.

58. The method of any one of claims 1 to 57, wherein the subject is further evaluated for one or more biomarkers associated with anti-tumor response.

59. A kit for treating cancer in a subject, comprising: a) a therapeutically effective amount of a compound represented by formula (I): or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, conformational isomer, tautomer or a combination thereof, or a compound represented by formula (10b): and b) a therapeutically effective amount of sotolacib, and instructions for effective administration.

60. The kit of claim 59, wherein the compound of formula (I) or (10b) and sotolacib are formulated for concomitant administration.

61. The kit of claim 59, wherein the compound of formula (I) or (10b) and sotolacib are formulated for sequential administration.

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