Methods and compositions for the treatment of KRAS mutant cancers

Abstract

The present disclosure provides methods and compositions for the treatment of KRAS mutant cancer.Aspects of the present disclosure are directed to the method for the treatment of KRAS mutant cancer, including KRAS mutant non-small cell lung cancer, comprising administering a KRAS inhibitor (e.g., sotrasib, adagrasib) and poziotinib.Also disclosed is a pharmaceutical composition comprising a KRAS inhibitor (e.g., sotrasib, adagrasib) and poziotinib.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 227,237, filed July 29, 2021, which is incorporated by reference in its entirety.

[0002] I. FIELD OF THE INVETION Aspects of the invention relate to at least the fields of cancer biology and medicine. [Background technology]

[0003] II. Background Novel KRAS G12C Early reports investigating KRAS inhibitors (e.g., sotrasib, adagrasib) G12C Within hours of inhibition, KRAS mutant cells upregulated HB-EGF, EGFR, and HER4 ligands, and HB-EGF upregulation was associated with increased expression of active KRAS. G12C In another study, inhibition of EGFR resulted in a transient attenuation of KRAS-mediated tumorigenesis in a preclinical model of KRAS mutant lung cancer. However, upregulation of other ErbB family members, including ErbB2 (HER2), ErbB3 (HER3), and ErbB4 (HER4), restored downstream signaling that attenuated the antitumor effect. Thus, KRAS G12C Inhibitor application limits the efficacy of treatment.

[0004] To improve the treatment of patients with KRAS mutant cancers G12C There is a need for methods and compositions to overcome the limited efficacy of KRAS inhibitors, including inhibitors. Summary of the Invention

[0005] overview Aspects of the present disclosure address certain needs in the field of cancer medicine by providing improved methods and compositions for the treatment of KRAS mutant cancer.Thus, in some aspects, a method of treating a subject for KRAS mutant cancer is provided herein, comprising administering a KRAS inhibitor and poziotinib to the subject.Also disclosed is a pharmaceutical composition comprising a KRAS inhibitor and poziotinib.In some embodiments, the KRAS inhibitor is a KRAS inhibitor such as sotrasib or adagrasib. G12C It is an inhibitor.

[0006] Embodiments of the disclosure include methods for treating a subject for KRAS mutant cancer, G12C Methods for Treating a Subject for KRAS Mutant Cancer G12C Methods for Treating a Subject for Non-Small Cell Lung Cancer, KRAS G12C Included are methods for detecting mutations, methods for diagnosing subjects with KRAS mutant cancer, and pharmaceutical compositions comprising a KRAS inhibitor and poziotinib. The methods of the present disclosure may include at least one, two, three, or more of the following steps: administering a KRAS inhibitor; G12C administering a selective HER2-4 inhibitor; administering poziotinib; administering a KRAS G12C administering a composition comprising an inhibitor and poziotinib, detecting a KRAS mutation in the subject, G12C Detecting the mutation, diagnosing the subject as having KRAS mutant cancer, and administering an additional cancer treatment. Any one or more of the foregoing steps may be excluded from the embodiments of the present disclosure. The pharmaceutical composition comprises a KRAS inhibitor, a KRAS G12C The composition may include one or more of the following inhibitors: sotrasib, a pharmaceutically acceptable salt of sotrasib, adagrasib, a pharmaceutically acceptable salt of adagrasib, poziotinib, a pharmaceutically acceptable salt of poziotinib, and a pharmaceutically acceptable excipient. Any one or more of the foregoing components may be excluded from embodiments of the present disclosure.

[0007] In some embodiments, disclosed herein are methods of treating a subject for KRAS mutant cancer, comprising administering to the subject an effective amount of (a) a KRAS inhibitor and (b) poziotinib. In some embodiments, the KRAS inhibitor and poziotinib are administered substantially simultaneously. In some embodiments, the KRAS inhibitor and poziotinib are administered sequentially. In some embodiments, the KRAS inhibitor is administered prior to administering poziotinib. In some embodiments, the KRAS inhibitor is administered after administering poziotinib. In some embodiments, the KRAS inhibitor is administered in combination with a KRAS inhibitor. G12C In some embodiments, the KRAS mutant cancer is KRAS mutant non-small cell lung cancer. In some embodiments, the KRAS mutant cancer is KRAS mutant colorectal cancer. In some embodiments, the KRAS mutant cancer is KRAS mutant pancreatic cancer.

[0008] In some embodiments, KRAS G12C Also disclosed herein are methods of treating a subject for non-small cell lung cancer, the methods comprising administering to a subject an effective amount of (a) a KRAS G12C In some embodiments, the method further comprises administering to the subject (a) a KRAS inhibitor and (b) poziotinib. G12C The KRAS inhibitor and poziotinib are administered substantially simultaneously. G12C The inhibitor and poziotinib are administered sequentially. G12C The KRAS inhibitor is administered prior to administering poziotinib. G12C The inhibitor is administered after administration of poziotinib.

[0009] In some embodiments, the method comprises detecting KRAS in a subject. G12C In some embodiments, the method further comprises detecting a KRAS mutation. G12C The inhibitor is sotorasib (AMG 510). G12CThe inhibitor is adagrasib (MRTX849). In some embodiments, the subject has been previously treated with a cancer therapy. In some embodiments, the cancer therapy has included chemotherapy. In some embodiments, the cancer therapy has included a KRAS inhibitor. In some embodiments, the subject has been determined to be resistant to the cancer therapy. In some embodiments, the subject has not been previously treated with a KRAS inhibitor. In some embodiments, poziotinib is administered at a dose of 0.1 mg to 50 mg. In some embodiments, poziotinib is administered at least TIFF2024527109000001.tif41170, at most TIFF2024527109000002.tif41170, approx. TIFF2024527109000003.tif41170, or exactly TIFF2024527109000004.tif41170, or any range or value derivable therein. In some embodiments, poziotinib is administered at a dose of 1 mg to 25 mg. In some embodiments, poziotinib is administered at a dose of 1 mg to 5 mg. In some embodiments, poziotinib is administered orally. In some embodiments, the KRAS inhibitor and poziotinib are administered at least 1, 2, 3, 4, 5, 6, or 7 times per day, at most 1, 2, 3, 4, 5, 6, or 7 times, or exactly 1, 2, 3, 4, 5, 6, or 7 times per day for multiple days. In some embodiments, the KRAS inhibitor and poziotinib are administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 times per week, or at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 times per week for multiple weeks. In some embodiments, the KRAS inhibitor and poziotinib are administered once per day for multiple days. In some embodiments, the KRAS inhibitor and poziotinib are administered twice a day for multiple days.In some embodiments, the method further comprises administering additional cancer treatment to the subject.In some embodiments, the additional cancer treatment comprises chemotherapy, radiation therapy, immunotherapy, or a combination thereof.

[0010] Also disclosed herein in some embodiments is a pharmaceutical composition comprising: (a) a KRAS inhibitor; (b) poziotinib; and (c) a pharma- ceutical acceptable excipient. In some embodiments, the KRAS inhibitor is G12C In some embodiments, the KRAS G12C The inhibitor is sotorasib (AMG 510). G12CThe inhibitor is adagrasib (MRTX849). In some embodiments, the poziotinib is at a dose of 0.1 mg to 50 mg. In some embodiments, the poziotinib is at least TIFF2024527109000005.tif41170, at most TIFF2024527109000006.tif41170, approx. TIFF2024527109000007.tif41170, or exactly TIFF2024527109000008.tif41170, or any range or value derivable therein. In some embodiments, poziotinib is at a dose of 1 mg to 25 mg. In some embodiments, poziotinib is at a dose of 1 mg to 5 mg.

[0011] In some embodiments, KRAS G12C Further disclosed herein is a method of treating a subject for non-small cell lung cancer, the method comprising administering to a subject an effective amount of (a) a KRAS G12C inhibitor; and (b) administering poziotinib at a dose of 1 mg to 5 mg twice daily for multiple days to the subject. In some embodiments, the poziotinib is at least TIFF2024527109000009.tif41159, at most TIFF2024527109000010.tif41159, approx. TIFF2024527109000011.tif41159, or exactly TIFF2024527109000012.tif41159, or at any range or value dose derivable therein.

[0012] Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for the method of measurement or quantification.

[0013] The use of the words "a" or "an," when used in conjunction with the term "comprising," can mean "one," but it is also consistent with the meaning of "one or more," "at least one," and "one or more than one."

[0014] The term "and / or" means "and" or "or." By way of example, A, B, and / or C includes A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C. In other words, "and / or" functions as an inclusive "or."

[0015] The words "comprising" (and any form of comprising, such as "comprise" and "comprises"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "includes" and "include"), or "containing" (and any form of containing, such as "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0016] The compositions and methods for their use may "comprise," "consist essentially of," or "consist" of any of the components or steps disclosed throughout this specification. Compositions and methods "consisting essentially of" any of the disclosed components or steps limit the claims to particular materials or steps that do not materially affect the basic or novel characteristics of the claimed invention.

[0017] "Individual," "subject," and "patient" are used interchangeably and can refer to a human or non-human.

[0018] Any method in the context of a therapeutic, diagnostic, or physiological purpose or effect may also be described in "use" claim language, such as the "use of" any compound, composition, or agent discussed herein to achieve or carry out the described therapeutic, diagnostic, or physiological purpose or effect.

[0019] It is expressly contemplated that any limitation discussed with respect to one aspect of the invention may be applied to any other aspect of the invention. Moreover, any composition of the invention may be used in any method of the invention, and any method of the invention may be used to produce or utilize any composition of the invention. Aspects of the embodiments shown in the examples are also embodiments that may be implemented in the context of embodiments discussed elsewhere in different examples, or elsewhere in this application, such as in the summary, detailed description, claims, and brief description of the figures.

[0020] Other objects, features, and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and specific examples, while indicating particular embodiments of the present invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. [Brief description of the drawings]

[0021] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. An understanding of the present invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein. [Figure 1]Figure 1A shows a Western blot obtained from 4 hours of treatment of H23 cells with the indicated concentrations of sotorasib (AMG 510) or DMSO control. Figure 1B shows sotorasib (AMG 510) IC50 values ​​in H358, H1378, H1792, and H2030 cells when combined with either 100 nM afatinib, 100 nM poziotinib, or DMSO control. Figure 1C shows adagrasib (MRTX849) IC50 values ​​in H358, H1378, H1792, and H2030 cells when combined with either 100 nM afatinib, 100 nM poziotinib, or DMSO control. [Diagram 2] 1 shows pEGFR and pHER2 levels in NSCLC cell lines harboring KRAS G12C mutations treated with sotorasib or adagrasib for 4 hours. [Diagram 3] Figure 3A shows phosphorylation of ERBB family members in HCC44, H2122, and H358 NSCLC cells (all harboring KRAS G12C mutations) treated with adagrasib for 72 hours. Figure 3B shows phosphorylation of ERBB family members in HCC44, H2122, and H358 NSCLC cells (all harboring KRAS G12C mutations) treated with sotorasib for 72 hours. [Figure 4] Figure 4A shows the sensitivity of Ba / F3 cells expressing EGFR, EGFR / HER2, HER2 / HER3, HER2 / HER4, HER3 / HER4, and HER4 to poziotinib. Figure 4B shows the sensitivity of Ba / F3 cells expressing EGFR, EGFR / HER2, HER2 / HER3, HER2 / HER4, HER3 / HER4, and HER4 to afatinib. [Diagram 5] Shows the selectivity of poziotinib for various receptors. [Figure 6] Figure 6A shows the resistance of H358 cells to treatment with a combination of adagrasib and exogenous EGF or NRG1. Figure 6B shows the resistance of H358 cells to treatment with a combination of sotorasib and exogenous EGF or NRG1. [Figure 7] Figure 7A shows the synergistic effect of treating H23, HCC44, H2122, and H1792 cells (NSCLC carrying KRAS G12C mutations) with sotorasib alone or in combination with poziotinib. Figure 7B shows the synergistic effect of treating H23, HCC44, H2122, and H1792 cells (NSCLC carrying KRAS G12C mutations) with adagrasib alone or in combination with poziotinib. [Figure 8] Figure 8A shows phosphorylation of ERBB family members in HCC44, H2122, and H358 NSCLC cells (all carrying KRAS G12C mutations) treated with sotrasib alone or in combination with afatinib or poziotinib. Figure 8B shows phosphorylation of ERBB family members in HCC44, H2122, and H358 NSCLC cells (all carrying KRAS G12C mutations) treated with adagrasib alone or in combination with afatinib or poziotinib. [Figure 9] Figure 9A shows the effect of sotrasib treatment alone or in combination with poziotinib (pozi) or afatinib (afat) on tumor volume in a PDX model of KRAS G12C mutant NSCLC. Figure 9B shows the effect of sotrasib treatment alone or in combination with poziotinib (pozi) or afatinib (afat) on progression-free survival in a PDX model of KRAS G12C mutant NSCLC. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] Detailed Description The present disclosure provides that administration of poziotinib, which is described herein to have selectivity for HER2-4 over EGFR, inhibits KRAS activation in KRAS mutant tumor cells. G12C The present disclosure is based, at least in part, on the surprising discovery that KRAS inhibitors (e.g., KRAS inhibitors such as sotrasib or adagrasib) synergistically enhance the cytotoxicity of the inhibitors. G12CThe present disclosure is directed to a method for treating a subject with KRAS mutant cancer, comprising administering a KRAS inhibitor and poziotinib. Also disclosed is a pharmaceutical composition comprising a KRAS inhibitor, poziotinib, and one or more pharma- ceutically acceptable excipients. Further aspects and embodiments of the disclosure are further described herein.

[0023] I. Treatment method Aspects of the present disclosure are directed to compositions comprising a therapeutically effective amount of one or more cancer therapeutics, and administering such compositions to a subject or patient in need thereof. In some embodiments, the one or more cancer therapeutics include a KRAS inhibitor and poziotinib.

[0024] The compositions of the present disclosure may be used for in vivo, in vitro, or ex vivo administration. The route of administration of the compositions may be, for example, intradermal, subcutaneous, intravenous, oral, topical, topical, and intraperitoneal administration.

[0025] The treatment provided herein may include the administration of a combination of a KRAS inhibitor and a therapeutic agent such as poziotinib.The treatment may be administered in any suitable manner known in the art.For example, the KRAS inhibitor and poziotinib may be administered sequentially (at different times) or simultaneously (at the same or about the same time; also referred to as "substantially simultaneously").In some embodiments, the KRAS inhibitor and poziotinib are administered in separate compositions.In some embodiments, the KRAS inhibitor and poziotinib are in the same composition.

[0026] In some embodiments, the KRAS inhibitor and poziotinib are administered substantially simultaneously. In some embodiments, the KRAS inhibitor and poziotinib are administered sequentially. In some embodiments, the KRAS inhibitor, poziotinib, and the additional cancer treatment are administered sequentially. In some embodiments, the KRAS inhibitor is administered before poziotinib is administered. In some embodiments, the KRAS inhibitor is administered after poziotinib is administered.

[0027] The embodiments of the present disclosure relate to compositions and methods, including therapeutic compositions. Different therapies may be administered in one composition, or in more than one composition, such as two compositions, three compositions, or four compositions. Various combinations of agents may be used.

[0028] The therapeutic agent of the present disclosure can be administered by the same or different administration route.In some embodiments, the therapeutic agent of the present disclosure (e.g., KRAS inhibitor, poziotinib) is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intracerebroventricularly, or intranasally.The appropriate dosage can be determined based on the type of disease to be treated, the severity and course of the disease, the individual's clinical condition, the individual's medical history and response to treatment, and the discretion of the attending physician.Different therapeutic agents can be administered by the same or different administration route.

[0029] Treatments may include various "unit doses". A unit dose is defined as containing a predetermined amount of a therapeutic composition. The amount to be administered, as well as the specific route and formulation, are within the skill of one of ordinary skill in the clinical arts in determining. In the case of intravenous administration, the unit dose need not be administered as a single injection, but may include a continuous infusion over a period of time. In some embodiments, the unit dose comprises a single administrable dose.

[0030] In some embodiments, the KRAS inhibitor is administered at a dose of 1 mg / kg to 5000 mg / kg. TIFF2024527109000013.tif238160, at most TIFF2024527109000014.tif238160, or approx. TIFF2024527109000015.tif238160, or any range or value derivable therein. In some embodiments, the KRAS inhibitor is administered at a dose of at least TIFF2024527109000016.tif77160, at most TIFF2024527109000017.tif77160, approx. TIFF2024527109000018.tif77160, or exactly TIFF2024527109000019.tif77160, or any range or value derivable therein.

[0031] In some embodiments, a single dose of poziotinib is administered. In some embodiments, multiple doses of poziotinib are administered. In some embodiments, poziotinib is administered in a single dose. TIFF2024527109000020.tif238160, at most TIFF2024527109000021.tif238160, or approx. TIFF2024527109000022.tif238160, or any range or value derivable therein. In some embodiments, poziotinib is administered at a dose of 0.1 mg / kg to 100 mg / kg.

[0032] In some embodiments, poziotinib comprises at least TIFF2024527109000023.tif77159, at most TIFF2024527109000024.tif77159, approx. TIFF2024527109000025.tif77159, or exactly TIFF2024527109000026.tif77159, or any range or value derivable therein. In some embodiments, poziotinib is administered at a dose of 0.1 mg to 50 mg. In some embodiments, poziotinib is administered at a dose of 1 mg to 25 mg. In some embodiments, poziotinib is administered at a dose of 1 mg to 5 mg.

[0033] The amount to be administered depends on the desired treatment effect, depending on both the number of treatments and the unit dose. It will be understood that an effective dose refers to the amount required to achieve a particular effect. Furthermore, such doses can be administered multiple times during a day and / or multiple days, weeks, or months. In some embodiments, the composition of the present disclosure (e.g., poziotinib and KRAS inhibitor) is administered to the subject 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times or more per day for multiple days. In some embodiments, poziotinib and KRAS inhibitor are administered to the subject once per day for multiple days. In some embodiments, poziotinib and KRAS inhibitor are administered to the subject twice per day for multiple days. In some embodiments, the compositions of the disclosure (e.g., poziotinib and a KRAS inhibitor) are administered to a subject 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 or more times per week for multiple weeks.

[0034] In certain embodiments, an effective dose of the pharmaceutical composition may provide a blood level of about 1 μM to 150 μM. In other embodiments, an effective dose provides a blood level of about 4 μM to 100 μM; or about 1 μM to 100 μM; or about 1 μM to 50 μM; or about 1 μM to 40 μM; or about 1 μM to 30 μM; or about 1 μM to 20 μM; or about 1 μM to 10 μM; or about 10 μM to 150 μM; or about 10 μM to 100 μM; or about 10 μM to 50 μM; or about 25 μM to 150 μM; or about 25 μM to 100 μM; or about 25 μM to 50 μM; or about 50 μM to 150 μM; or about 50 μM to 100 μM (or any range derivable therein). In other embodiments, the dose may provide the following blood levels of the agent resulting from the therapeutic agent being administered to the subject: TIFF2024527109000027.tif26166, at least approx. TIFF2024527109000028.tif26166, or at most about TIFF2024527109000029.tif26166, or any range derivable therein. In certain embodiments, a therapeutic agent administered to a subject is metabolized in the body to a metabolized therapeutic agent, in which case blood levels may refer to the amount of that agent. Alternatively, to the extent that the therapeutic agent is not metabolized by the subject, blood levels discussed herein may refer to a non-metabolized therapeutic agent.

[0035] In some embodiments, poziotinib is administered to a subject in an amount of 0.1 mg to 50 mg, or any range or value derivable therein. Poziotinib can be administered in chloride salt form and can be administered orally, such as in a tablet. Poziotinib can be administered in a dose of 1 to 25 mg, such as a dose of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 mg, or any range or value derivable therein. Dosing can be twice daily, daily, every other day, every third day, or weekly. In some embodiments, dosing is twice per day. Dosing can be on a continuous schedule, such as on a 28-day cycle.

[0036] The precise amount of therapeutic composition depends on the judgment of the health care practitioner and is peculiar to each individual. Factors influencing the dosage include the physical and clinical condition of the patient, the route of administration, the intended purpose of the treatment (alleviation of symptoms versus cure), and the efficacy, stability, and toxicity of the particular therapeutic agent or other treatments the subject may be undergoing.

[0037] It will be understood and appreciated by those skilled in the art that dosage units of μg / kg or mg / kg of body weight can be converted and expressed in equivalent concentration units (blood levels) of μg / ml or mM, such as 4 μM to 100 μM. It will also be understood that uptake is species and organ / tissue dependent. Applicable conversion factors and physiological assumptions to be considered for uptake and concentration measurements are well known and will enable those skilled in the art to convert one concentration measurement to another and make reasonable comparisons and conclusions regarding the doses, potencies, and results described herein.

[0038] In certain instances, it may be desirable to administer multiple doses of the composition, for example, 2, 3, 4, 5, 6, or more doses. Administration may be at intervals of 1, 2, 3, 4, 5, 6, 7, 8 to 5, 6, 7, 8, 9, 10, 11, or 12 weeks (including all ranges therebetween).

[0039] The phrase "pharmacologically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not cause adverse, allergic or other untoward reactions when administered to animals or humans. As used herein, "pharmacologically acceptable carriers" include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmacoactive substances is well known in the art. As long as any conventional media or agent is not incompatible with the active ingredient, its use in immunogenic and therapeutic compositions is contemplated. Other supplementary active ingredients, such as anti-infective agents and vaccines, can also be included in the composition.

[0040] Active compound can be formulated for parenteral administration, for example, can be formulated for injection via intravenous, intramuscular, subcutaneous or intraperitoneal route.Typically, such compositions can be prepared as either liquid solution or suspension;solid forms can also be prepared that are suitable for use in preparing solution or suspension by adding liquid before injection;preparation can also be emulsified.

[0041] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations, for example, containing aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that it can be easily squirted. It must also be stable under the conditions of manufacture and storage, and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

[0042] The pharmaceutical composition may contain a solvent or dispersion medium, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include an isotonic agent, for example, sugar or sodium chloride. Prolonged absorption of the injectable composition can be brought about by the use of absorption-delaying agents in the composition, for example, aluminum monostearate and gelatin.

[0043] Sterile injectable solution is prepared by incorporating the required amount of active compound into a suitable solvent with various other ingredients as listed above, as necessary, and then sterilizing by filtration or equivalent procedures.Generally, dispersion is prepared by incorporating various sterilized active ingredients into a sterile vehicle that contains a basic dispersion medium and other necessary ingredients from those listed above.In the case of sterile powder for preparing sterile injectable solution, the preferred method of preparation is vacuum drying and freeze-drying technique, which obtains a powder of active ingredient with any additional desired ingredients from its pre-sterilized filtered solution.

[0044] The administration of the composition is typically via any common route, including, but not limited to, oral administration and / or intravenous administration. Alternatively, administration can be by orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal, or intranasal administration. Such compositions will usually be administered as pharma- ceutically acceptable compositions, including physiologically acceptable carriers, buffers, or other excipients.

[0045] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutically or prophylactically effective.

[0046] A. Cancer Treatment In some embodiments, the method includes administering a cancer treatment to the patient. The cancer treatment can be selected based on the expression level measurement, alone or in combination with a clinical risk score calculated for each patient. In some embodiments, the cancer treatment includes a local cancer treatment. In some embodiments, the cancer treatment excludes a systemic cancer treatment. In some embodiments, the cancer treatment excludes a local treatment. In some embodiments, the cancer treatment includes a local cancer treatment without administration of a systemic cancer treatment. In some embodiments, the cancer treatment includes an immunotherapy, which can be an immune checkpoint therapy. In some embodiments, the cancer treatment is a KRAS inhibitor. In some embodiments, the cancer treatment is poziotinib. Any of these cancer treatments can be excluded. A combination of these treatments can also be administered. For example, as disclosed herein, a combination of a KRAS inhibitor and poziotinib is synergistically effective in treating KRAS mutant cancers; thus, aspects of the disclosure are directed to cancer treatments including a combination of a KRAS inhibitor (e.g., sotrasib or adagrasib) and poziotinib.

[0047] The term "cancer" as used herein can be used to describe solid tumors, metastatic cancers, or non-metastatic cancers. In certain embodiments, cancer can occur in the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, duodenum, small intestine, large intestine, colon, rectum, anus, gums, head, kidney, liver, lung, nasopharynx, cervix, ovary, pancreas, prostate, skin, stomach, testicle, tongue, or uterus. In some embodiments, the cancer is a recurrent cancer. In some embodiments, the cancer is a stage I cancer. In some embodiments, the cancer is a stage II cancer. In some embodiments, the cancer is a stage III cancer. In some embodiments, the cancer is a stage IV cancer.

[0048] The cancer may specifically be of the following histological types, but is not limited to: neoplasm, malignant; carcinoma; carcinoma, unclassified; giant cell and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; mixed hepatocellular and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyps; adenocarcinoma, familial polyposis coli; solid tumors; carcinoid tumor, malignant; bronchioloalveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophilic carcinoma; acidophilic adenocarcinoma; basophilic carcinoma; clear cell Adenocarcinoma;Granular cell carcinoma;Follicular adenocarcinoma;Papillary and follicular adenocarcinoma;Nonencapsulated sclerosing carcinoma;Adrenal cortical carcinoma;Endometrioid carcinoma;Cutaneous adnexal carcinoma;Apocrine adenocarcinoma;Sebaceous gland adenocarcinoma;Earwax adenocarcinoma;Mucoepidermoid carcinoma;Cystadenocarcinoma;Papillary cystadenocarcinoma;Papillary serous cystadenocarcinoma;Mucinous cystadenocarcinoma;Mucinous adenocarcinoma;Signet ring cell carcinoma;Invasive ductal carcinoma;Medullary carcinoma;Lobular carcinoma;Inflammatory carcinoma;Paget's disease, breast;Acinic cell carcinoma;Adenosquamous carcinoma;Adenocarcinoma with squamous metaplasia;Thymoma, malignant;Ovarian stromal tumor, malignant;Theca cell tumor, malignant;Granulosa cell tumor, malignant;Androblastoma, malignant;Sertoli cell tumor;Leydig cell tumor, malignant;Lipid cell tumor alveolar tumor, malignant;paraganglioma, malignant;extramammary paraganglioma, malignant;pheochromocytoma;glomus tumor;malignant melanoma;amelanotic melanoma;superficial spreading melanoma;malignant melanoma in giant pigmented nevus;epithelioid cell melanoma;blue nevus, malignant;sarcoma;fibrosarcoma;fibrous histiocytoma, malignant;myxosarcoma;liposarcoma;leiomyosarcoma;rhabdomyosarcoma;embryonal rhabdomyosarcoma;alveolar rhabdomyosarcoma;stromal sarcoma;mixed tumor, malignant;mixed Müllerian tumor;nephroblastoma;hepatoblastoma;carcinosarcoma;mesenchymoma, malignant;Brenner tumor, malignant;phyllodes tumor, malignant;synovial sarcoma;mesothelioma, malignant;dysgerminoma;embryonal carcinoma;teratoma , malignant;ovarian goiter, malignant;choriocarcinoma;mesonephroma, malignant;angiosarcoma;hemangioendothelioma, malignant;Kaposi's sarcoma;hemangiopericytoma, malignant;lymphangiosarcoma;osteosarcoma;parosteal osteosarcoma;chondrosarcoma;chondroblastoma, malignant;mesenchymal chondrosarcoma;giant cell tumor of bone;Ewing's sarcoma;odontogenic tumor, malignant;ameloblastoma;ameloblastoma, malignant;ameloblastic fibrosarcoma;pinealoma, malignant;chordoma;glioma, malignant;ependymoma;astrocytoma;protoplasmic astrocytoma;fibrous astrocytoma;astroblastoma;glioblastoma;oligodendroglioma;oligodendroglioma;primitive neuroectodermal;cerebellar sarcoma;ganglioneoblastoma;neuroblastoma;Retinoblastoma; Olfactory nerve tumor; Meningioma, malignant; Neurofibrosarcoma; Schwannoma, malignant; Granular cell tumor, malignant; Malignant lymphoma; Hodgkin's disease; Hodgkin's; Side granuloma; Malignant lymphoma, small lymphocytic; Malignant lymphoma, large cell, diffuse; Malignant lymphoma, follicular; Mycosis fungoides; Other specific non-Hodgkin's lymphoma; Malignant histiocytosis; Multiple myeloma; Mast cell sarcoma; Immunoproliferative small intestinal disease; Leukemia; Lymphocytic leukemia; Plasma cell leukemia; Erythroleukemia; Lymphosarcoma cell leukemia; Myeloid leukemia; Basophilic leukemia; Eosinophilic leukemia; Monocytic leukemia; Mast cell leukemia; Megakaryoblastic leukemia; Myeloid sarcoma; and Hairy cell leukemia. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is a KRAS mutant cancer. In some embodiments, the KRAS mutant cancer is a KRAS G12C mutant cancer;

[0049] In some embodiments, the method of the present disclosure includes treating a subject suffering from cancer (e.g., KRAS mutant cancer) by administering a therapeutically effective amount of a KRAS inhibitor and poziotinib. As used herein, the term "therapeutically effective amount" is synonymous with "effective amount," "therapeutically effective dose," and / or "effective dose," and refers to an amount of an agent (or combination of agents) sufficient to obtain a desired result or to have a desired effect on the particular condition being treated. In some embodiments, a therapeutically effective amount is an amount sufficient to improve at least one symptom, behavior, or event associated with a pathological, abnormal, or otherwise undesirable condition, or an amount sufficient to prevent or reduce the probability of such a condition occurring or recurring, or an amount sufficient to delay the worsening of such a condition. For example, in some embodiments, an effective amount refers to an amount of a KRAS inhibitor and poziotinib in combination that can treat or prevent cancer in a subject. An effective amount can vary depending on the organism or individual being treated. The appropriate effective amount to be administered for a particular application of the method of the present disclosure can be determined by one of skill in the art using the guidance provided herein. As used herein, the term "treatment", "treat" or "treating" refers to an intervention that attempts to change the natural course of the subject being treated, and can be carried out either for prevention or during the course of the pathology of disease or condition.Treatment can be useful in achieving one or more of various desired outcomes, including, for example, preventing the occurrence or recurrence of disease, alleviating or reducing the severity of symptoms, and alleviating any direct or indirect pathological consequences of disease, preventing disease spread, slowing down the rate of disease progression, improving or remission of disease state, and remission or improving prognosis.

[0050] 1. KRAS mutant cancer Aspects of the present disclosure are directed to methods for the treatment of KRAS mutant cancer. As used herein, "KRAS mutant cancer" describes a cancer that carries one or more KRAS mutations. Thus, a subject with KRAS mutant cancer describes a subject with cancer, where cancer cells from the subject are identified to have a KRAS mutation. KRAS mutations include, for example, G12 mutations (e.g., G12A, G12C, G12D, G12R, G12V), G13 mutations (e.g., G13D), and Q61 mutations (e.g., Q61K, Q61L, Q61H). In some embodiments, the subject of the present disclosure has KRAS G12C mutant non-small cell lung cancer (NSCLC). In some embodiments, the subject of the present disclosure has KRAS G12C mutant colorectal cancer. In some embodiments, the subject of the present disclosure has KRAS G12C mutant pancreatic cancer.

[0051] B. KRAS Inhibitors Aspects of the present disclosure include KRAS inhibitors and methods of use thereof. As used herein, "KRAS inhibitor" describes any molecule capable of inhibiting the activity and / or reducing the expression of the GTPase KRas ("KRAS" or "K-Ras") protein. In some embodiments, the KRAS inhibitor is an oligonucleotide capable of reducing the expression of the KRAS protein in a cell. In some embodiments, the KRAS inhibitor is an inhibitor of KRAS enzymatic activity. In some embodiments, the KRAS inhibitor is a molecule capable of inactivating KRAS by trapping KRAS in a GDP-bound state. In some embodiments, the KRAS protein targeted by the KRAS inhibitors of the present disclosure is a mutant KRAS protein. Mutant KRAS proteins include KRAS proteins with a G12 mutation, such as, for example, G12A, G12C, G12D, G12R, or G12V. In some embodiments, the mutant KRAS protein is a KRAS protein with a G12C mutation ("KRAS"). G12C ").

[0052] In some embodiments, the KRAS inhibitors of the present disclosure are G12C As used herein, "KRAS" is a G12C Inhibitors G12C In some embodiments, the present invention describes any molecule capable of inhibiting the activity and / or reducing the expression of KRAS. G12C Inhibitors include KRAS G12C In some embodiments, the compound is capable of trapping KRAS in a GDP-bound state, thereby inhibiting the enzymatic activity of the protein. G12C Inhibitors inhibit KRAS compared to wild-type KRAS protein. G12C In some embodiments, the protein is preferentially inhibited. G12C The inhibitor does not inhibit the wild-type KRAS protein. G12C Examples of inhibitors include, but are not limited to, ARS-1620, ARS-853, sotorasib (AMG 510), and adagrasib (MRTX849). G12C The inhibitor is 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-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, i.e., sotorasib, having a structure represented by formula I, or a pharma- ceutical acceptable salt thereof. G12C The inhibitor is {(2S)-4-[7-(8-chloronaphthalen-1-yl)-2-{[(2S)-1-methylpyrrolidin-2-yl]methoxy}-5,6,7,8 tetrahydropyrido[3,4-d]pyrimidin-4-yl]-1-(2-fluoroprop-2-enoyl)piperazin-2-yl}acetonitrile, having the structure represented by Formula III, i.e., adagrasib, or a pharma-ceutically acceptable salt thereof. TIFF2024527109000030.tif59128

[0053] C. Poziotinib 1-[4-[4-(3,4-dichloro-2-fluoroanilino)-7-methoxyquinazolin-6-yl]oxypiperidin-1-yl]prop-2-en-1-one, or poziotinib (also referred to as "HM781-36" or "HM781-36B"), has the formula III: It is a compound having a structure represented by TIFF2024527109000031.tif71128.

[0054] Poziotinib is a pan-HER inhibitor capable of inhibiting the activity of ErbB1 (EGFR), ErbB2 (HER2), ErbB3 (HER3), and ErbB4 (HER4). Poziotinib is described, for example, in PCT Publication No. WO 2020 / 005932 and Cha MY,. et al,. Int J Cancer. 2012 May 15;130(10):2445-54, each of which is incorporated herein by reference in its entirety. Disclosed herein are compositions comprising poziotinib or a pharma- ceutically acceptable salt thereof. Also disclosed are methods for the use of poziotinib, optionally in combination with one or more KRAS inhibitors, for the treatment of KRAS mutant cancer. As used herein, compositions and methods comprising "poziotinib" describe compositions and methods comprising a compound having a structure represented by formula III or a pharma- ceutically acceptable salt thereof.

[0055] D. Cancer Immunotherapy In some embodiments, the method includes the administration of cancer immunotherapy as a therapeutic agent. Cancer immunotherapy (sometimes called cancer immunology, abbreviated as IO) is the use of the immune system to treat cancer. Immunotherapies can be classified as active, passive, or hybrid (active and passive). These approaches take advantage of the fact that cancer cells often have molecules on their surface that can be detected by the immune system, known as tumor-associated antigens (TAA); they are often proteins or other macromolecules (e.g., carbohydrates). Active immunotherapy directs the immune system to attack tumor cells by targeting TAAs. Passive immunotherapy enhances existing anti-tumor responses and includes the use of monoclonal antibodies, lymphocytes, and cytokines. A variety of immunotherapies are known in the art, and examples are provided below.

[0056] 1. Checkpoint Inhibitors and Combination Treatments Aspects of the present disclosure may include administration of immune checkpoint inhibitors, examples of which are further described below. As disclosed herein, "checkpoint inhibitor therapy" (also "immune checkpoint blockade therapy", "immune checkpoint therapy", "ICT", "checkpoint blockade immunotherapy", or "CBI") refers to cancer therapy that includes providing one or more immune checkpoint inhibitors to a subject suffering from or suspected of having cancer.

[0057] PD-1 may act in the tumor microenvironment where T cells encounter infection or tumor. Activated T cells upregulate PD-1 and continuously express it in peripheral tissues. Cytokines such as IFN-gamma induce the expression of PDL1 on epithelial and tumor cells. PDL2 is expressed on macrophages and dendritic cells. The main role of PD-1 is to limit the activity of effector T cells in the periphery and prevent excessive damage to tissues during immune responses. The inhibitors of the present disclosure may block one or more of PD-1 and / or PDL1 activities.

[0058] Alternative names for "PD-1" include CD279 and SLEB2. Alternative names for "PDL1" include B7-H1, B7-4, CD274, and B7-H. Alternative names for "PDL2" include B7-DC, Btdc, and CD273. In some embodiments, PD-1, PDL1, and PDL2 are human PD-1, PDL1, and PDL2.

[0059] In some embodiments, the PD-1 inhibitor is a molecule that inhibits PD-1 from binding to its ligand binding partner. In certain aspects, the PD-1 ligand binding partner is PDL1 and / or PDL2. In another embodiment, the PDL1 inhibitor is a molecule that inhibits PDL1 from binding to its binding partner. In certain aspects, the PDL1 binding partner is PD-1 and / or B7-1. In another embodiment, the PDL2 inhibitor is a molecule that inhibits PDL2 from binding to its binding partner. In certain aspects, the PDL2 binding partner is PD-1. The inhibitor can be an antibody, an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide. Exemplary antibodies are described in U.S. Patent Nos. 8,735,553, 8,354,509, and 8,008,449, all of which are incorporated herein by reference. Other PD-1 inhibitors for use in the methods and compositions provided herein are known in the art, such as those described in U.S. Patent Application Nos. US2014 / 0294898, US2014 / 022021, and US2011 / 0008369, all of which are incorporated herein by reference.

[0060] In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody). In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, and pidilizumab. In some embodiments, the PD-1 inhibitor is an immunoadhesin (e.g., an immunoadhesin comprising an extracellular portion or a PD-1 binding portion of PDL1 or PDL2 fused to a constant region (e.g., an Fc region of an immunoglobulin sequence). In some embodiments, the PDL1 inhibitor comprises AMP-224. Nivolumab, also known as MDX-1106-04, MDX-1106, ONO-4538, BMS-936558, and OPDIVO®, is an anti-PD-1 antibody described in WO2006 / 121168. MK-3475, Merck3475, lambrolizumab, KEYTRUDA Pembrolizumab, also known as CT-011, hBAT, or hBAT-1, is an anti-PD-1 antibody described in WO2009 / 114335. Pidilizumab, also known as CT-011, hBAT, or hBAT-1, is an anti-PD-1 antibody described in WO2009 / 101611. AMP-224, also known as B7-DCIg, is a PDL2-Fc fusion soluble receptor described in WO2010 / 027827 and WO2011 / 066342. Additional PD-1 inhibitors include MEDI0680, also known as AMP-514, and REGN2810.

[0061] In some embodiments, the immune checkpoint inhibitor is a PDL1 inhibitor, such as durvalumab, also known as MEDI4736, atezolizumab, also known as MPDL3280A, avelumab, also known as MSB00010118C, MDX-1105, BMS-936559, or a combination thereof. In certain aspects, the immune checkpoint inhibitor is a PDL2 inhibitor, such as rHIgM12B7.

[0062] In some embodiments, the inhibitor comprises the heavy and light chain CDRs or VRs of nivolumab, pembrolizumab, or pidilizumab. Thus, in one embodiment, the inhibitor comprises the CDR1, CDR2, and CDR3 domains of the VH region of nivolumab, pembrolizumab, or pidilizumab, and the CDR1, CDR2, and CDR3 domains of the VL region of nivolumab, pembrolizumab, or pidilizumab. In another embodiment, the antibody competes for binding and / or binds to the same epitope on PD-1, PDL1, or PDL2 as the above-mentioned antibody. In another embodiment, the antibody has at least about 70, 75, 80, 85, 90, 95, 97, or 99% (or any range derivable therein) of variable region amino acid sequence identity with the above-mentioned antibody.

[0063] Another immune checkpoint that can be targeted in the methods provided herein is cytotoxic T lymphocyte-associated protein 4 (CTLA-4), also known as CD152. The complete cDNA sequence of human CTLA-4 has GenBank accession number L15006. CTLA-4 is found on the surface of T cells and acts as an "off" switch when it binds to B7-1 (CD80) or B7-2 (CD86) on the surface of antigen-presenting cells. CTLA4 is a member of the immunoglobulin superfamily that is expressed on the surface of helper T cells and transmits inhibitory signals to T cells. CTLA4 is similar to CD28, a T cell costimulatory protein, and both molecules bind to B7-1 and B7-2 on antigen-presenting cells. CTLA-4 transmits inhibitory signals to T cells, while CD28 transmits stimulatory signals. Intracellular CTLA-4 is also found in regulatory T cells and may be important for their function. T cell activation via T cell receptor and CD28 increases the expression of CTLA-4, an inhibitory receptor for B7 molecules. The inhibitor of the present disclosure can block one or more functions of CTLA-4, B7-1, and / or B7-2 activity. In some embodiments, the inhibitor blocks the interaction between CTLA-4 and B7-1. In some embodiments, the inhibitor blocks the interaction between CTLA-4 and B7-2.

[0064] In some embodiments, the immune checkpoint inhibitor is an anti-CTLA-4 antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody), an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide.

[0065] Anti-human CTLA-4 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present method can be generated using methods well known in the art. Alternatively, art-recognized anti-CTLA-4 antibodies can be used. For example, the anti-CTLA-4 antibodies disclosed in US 8,119,129, WO 01 / 14424, WO 98 / 42752; WO 00 / 37504 (CP675,206, also known as tremelimumab; formerly ticilimumab), US Patent No. 6,207,156; Hurwitz et al., 1998 can be used in the methods disclosed herein. The teachings of each of the aforementioned publications are incorporated herein by reference. Antibodies that compete with any of these art-recognized antibodies for binding to CTLA-4 can also be used. For example, humanized CTLA-4 antibodies are described in International Patent Application Nos. WO2001 / 014424, WO2000 / 037504, and US Pat. No. 8,017,114; all of which are incorporated herein by reference.

[0066] An additional anti-CTLA-4 antibody useful as a checkpoint inhibitor in the methods and compositions of the disclosure is ipilimumab (also known as 10D1, MDX-010, MDX-101, and Yervoy®) or antigen-binding fragments and variants thereof (see, e.g., WO 01 / 14424).

[0067] In some embodiments, the inhibitor comprises the heavy and light chain CDRs or VRs of tremelimumab or ipilimumab. Thus, in one embodiment, the inhibitor comprises the CDR1, CDR2, and CDR3 domains of the VH region of tremelimumab or ipilimumab and the CDR1, CDR2, and CDR3 domains of the VL region of tremelimumab or ipilimumab. In another embodiment, the antibody competes for binding to and / or binds to the same epitope on PD-1, B7-1, or B7-2 as the above-mentioned antibody. In another embodiment, the antibody has at least about 70, 75, 80, 85, 90, 95, 97, or 99% (or any range derivable therein) of variable region amino acid sequence identity with the above-mentioned antibody.

[0068] Another immune checkpoint that can be targeted in the methods provided herein is lymphocyte activation gene 3 (LAG3), also known as CD223 and lymphocyte activation 3. The complete mRNA sequence of human LAG3 has GenBank accession number NM_002286. LAG3 is a member of the immunoglobulin superfamily found on the surface of activated T cells, natural killer cells, B cells, and plasmacytoid dendritic cells. The primary ligand of LAG3 is MHC class II, which has been reported to negatively regulate T cell proliferation, activation, and homeostasis in a manner similar to CTLA-4 and PD-1, and to play a role in Treg suppressive function. LAG3 also helps maintain CD8+ T cells in a tolerogenic state and works with PD-1 to help maintain CD8 exhaustion during chronic viral infection. LAG3 is also known to be involved in dendritic cell maturation and activation. The inhibitors of the present disclosure can block one or more functions of LAG3 activity.

[0069] In some embodiments, the immune checkpoint inhibitor is an anti-LAG3 antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody), an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide.

[0070] Anti-human LAG3 antibody (or VH and / or VL domain derived therefrom) suitable for use in this method can be generated using methods well known in the art.Alternatively, art-recognized anti-LAG3 antibody can be used.For example, anti-LAG3 antibody can include GSK2837781, IMP321, FS-118, Sym022, TSR-033, MGD013, BI754111, AVA-017, or GSK2831781. US 9,505,839 (BMS-986016, also known as leratolimab); US 10,711,060 (IMP-701, also known as LAG525); US 9,244,059 (IMP731, also known as H5L7BW); US 10,344,089 (25F7, also known as LAG3.1); WO 2016 / 028672 (MK-4280, also known as 28G-10); WO 2017 / 019894 (BAP050); Burova E., et al., J. ImmunoTherapy Cancer, 2016; 4(Supp. 1): P195 (REGN3767); Yu, X., et al., mAbs, 2019; 11:6 The anti-LAG3 antibodies disclosed in (LBL-007) may be used in the methods disclosed herein.These and other anti-LAG-3 antibodies useful in the claimed disclosure are described in, e.g., WO 2016 / 028672, WO 2017 / 106129, WO 2017062888, WO 2009 / 044273, WO 2018 / 069500, WO 2016 / 126858, WO 2014 / 179664, WO 2016 / 200782, WO 2015 / 200119, WO 2017 / 019846, WO 2017 / 198741, WO 2017 / 220555, WO 2017 / 220569, WO 2018 / 071500, WO 2017 / 015560; WO 2017 / 025498, WO 2017 / 087589, WO 2017 / 087901, WO 2018 / 083087, WO 2017 / 149143, WO 2017 / 219995, US 2017 / 0260271, WO 2017 / 086367, WO 2017 / 086419, WO 2018 / 034227, and WO 2014 / 140180. The teachings of each of the aforementioned publications are incorporated herein by reference. Antibodies that compete with any of these art-recognized antibodies for binding to LAG3 can also be used.

[0071] In some embodiments, the inhibitor comprises the heavy and light chain CDRs or VRs of an anti-LAG3 antibody. Thus, in one embodiment, the inhibitor comprises the CDR1, CDR2, and CDR3 domains of the VH region of an anti-LAG3 antibody and the CDR1, CDR2, and CDR3 domains of the VL region of an anti-LAG3 antibody. In another embodiment, the antibody has at least about 70, 75, 80, 85, 90, 95, 97, or 99% (or any range derivable therein) variable region amino acid sequence identity with the above-mentioned antibody.

[0072] Another immune checkpoint that can be targeted in the methods provided herein is T cell immunoglobulin and mucin domain-containing-3 (TIM-3), also known as Hepatitis A Virus Cellular Receptor 2 (HAVCR2) and CD366. The complete mRNA sequence of human TIM-3 has GenBank accession number NM_032782. TIM-3 is found on the surface of IFNγ-producing CD4+ Th1 and CD8+ Tc1 cells. The extracellular region of TIM-3 consists of a single variable immunoglobulin domain (IgV) distal to the membrane and a glycosylated mucin domain of variable length located closer to the membrane. TIM-3 is an immune checkpoint that mediates T cell exhaustion together with other inhibitory receptors including PD-1 and LAG3. TIM-3 has also been shown to be a CD4+ Th1-specific cell surface protein that regulates macrophage activation. The inhibitors of the present disclosure can block one or more functions of TIM-3 activity.

[0073] In some embodiments, the immune checkpoint inhibitor is an anti-TIM-3 antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody), an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide.

[0074] Anti-human TIM-3 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present method can be generated using methods well known in the art. Alternatively, art-recognized anti-TIM-3 antibodies can be used. For example, anti-TIM-3 antibodies including MBG453, TSR-022 (also known as covolimab), and LY3321367 can be used in the methods disclosed herein. These and other anti-TIM-3 antibodies useful in the claimed disclosure can be found, for example, in US 9,605,070, US 8,841,418, US2015 / 0218274, and US 2016 / 0200815. The teachings of each of the aforementioned publications are incorporated herein by reference. Antibodies that compete with any of these art-recognized antibodies for binding to LAG3 can also be used.

[0075] In some embodiments, the inhibitor comprises the heavy and light chain CDRs or VRs of an anti-TIM-3 antibody. Thus, in one embodiment, the inhibitor comprises the CDR1, CDR2, and CDR3 domains of the VH region of an anti-TIM-3 antibody and the CDR1, CDR2, and CDR3 domains of the VL region of an anti-TIM-3 antibody. In another embodiment, the antibody has at least about 70, 75, 80, 85, 90, 95, 97, or 99% (or any range derivable therein) variable region amino acid sequence identity with the above-mentioned antibodies.

[0076] E. Oncolytic Viruses In some aspects, the cancer treatment of the present disclosure comprises oncolytic viruses. Oncolytic viruses are viruses that preferentially infect and kill cancer cells. As infected cancer cells are destroyed by oncolysis, they release new infectious virus particles or virions to help destroy remaining tumors. Oncolytic viruses are believed to not only cause direct destruction of tumor cells, but also stimulate host anti-tumor immune responses for long-term immunotherapy.

[0077] F. Chemotherapy In some embodiments, the cancer treatment of the present disclosure includes chemotherapy. Suitable classes of chemotherapeutic agents include (a) alkylating agents such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil), ethylenimines and methylmelamines (e.g., hexamethylmelamine, thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomustine, chlorozoticin, streptozocin), and triazines (e.g., dicarbazine); (b) folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., 5-fluorouracil, floxuridine, cytarabine, azauridine), and purine analogs and related substances (e.g., 6-mercaptopurine, 6-thioguam, benzodiazepine, benzoic acid ... (c) natural products such as vinca alkaloids (e.g., vinblastine, vincristine), epipodophyllotoxins (e.g., etoposide, teniposide), antibiotics (e.g., dactinomycin, daunorubicin, doxorubicin, bleomycin, plicamycin, and mitoxantrone), enzymes (e.g., L-asparaginase), and biological response modifiers (e.g., interferon-α), and (d) miscellaneous agents such as platinum coordination complexes (e.g., cisplatin, carboplatin, oxaliplatin), substituted ureas (e.g., hydroxyurea), methylhydiazine derivatives (e.g., procarbazine), and adrenal cortex suppressants (e.g., taxol and mitotane). In some embodiments, cisplatin is a particularly suitable chemotherapeutic agent.

[0078] Cisplatin is widely used to treat cancers, such as metastatic testicular or ovarian cancer, advanced bladder cancer, head or neck cancer, cervical cancer, lung cancer, or other tumors. Cisplatin is not absorbed orally and must therefore be delivered via other routes, such as intravenous, subcutaneous, intratumoral, or intraperitoneal injection. Cisplatin is administered at approximately 15 mg / m2 for 5 days every 3 weeks for a total of three courses, which is contemplated in certain embodiments. 2 ~about 20 mg / m 2 These may be used alone or in combination with other agents, using effective doses used in clinical applications, including:

[0079] Other suitable chemotherapeutic agents include microtubule inhibitors, such as paclitaxel ("Taxol") and doxorubicin hydrochloride ("doxorubicin"). Doxorubicin is poorly absorbed and is therefore preferably administered intravenously. In certain embodiments, a suitable intravenous dose for an adult is about 60 mg / m2 at intervals of about 21 days. 2 ~about 75 mg / m 2 or about 25 mg / m2 on each of 2 or 3 consecutive days repeated at intervals of about 3 to about 4 weeks. 2 ~about 30 mg / m 2 or approximately 20 mg / m once weekly 2 The lowest doses should be used in elderly patients if previous myelosuppression caused by previous chemotherapy or neoplastic bone marrow infiltration is present, or if the drug is combined with other myelopoietin-suppressing drugs.

[0080] Nitrogen mustard is another suitable chemotherapeutic agent useful in the methods of the present disclosure. Nitrogen mustards may include, but are not limited to, mechlorethamine (HN2), cyclophosphamide and / or ifosfamide, melphalan (L-sarcolysin), and chlorambucil. Cyclophosphamide (CYTOXAN® available from Mead Johnson, NEOSTAR® available from Adria) is another suitable chemotherapeutic agent. Suitable oral doses for adults include, for example, about 1 mg / kg / day to about 5 mg / kg / day, and intravenous doses include, for example, about 40 mg / kg to about 50 mg / kg initially in divided doses over a period of about 2 to about 5 days, or about 10 mg / kg to about 15 mg / kg every about 7 to about 10 days, or about 3 mg / kg to about 5 mg / kg twice weekly, or about 1.5 mg / kg / day to about 3 mg / kg / day. Due to adverse gastrointestinal effects, the intravenous route is preferred. Drugs may also be administered intramuscularly, by osmosis, or into body cavities.

[0081] Additional suitable chemotherapeutic agents include pyrimidine analogs such as cytarabine (cytosine arabinoside), 5-fluorouracil (fluorouracil; 5-FU), and floxuridine (fluorodeoxyuridine; FudR). 5-FU can be administered to a subject at a dosage anywhere between about 7.5 and about 1000 mg / m2. Furthermore, the dosing schedule of 5-FU can be over a variety of periods, for example, up to 6 weeks, or as determined by one of ordinary skill in the art to which this disclosure pertains.

[0082] Gemcitabine diphosphate (GEMZAR®, Eli Lilly & Co., "gemcitabine") is another suitable chemotherapeutic agent that is recommended for the treatment of advanced and metastatic pancreatic cancer and therefore may also be useful in certain embodiments of the present disclosure for these cancers.

[0083] The amount of chemotherapeutic agent delivered to the patient may vary. In one suitable embodiment, the chemotherapeutic agent may be administered in an amount effective to cause arrest or regression of cancer in the host when the chemotherapy is administered with the construct. In other embodiments, the chemotherapeutic agent may be administered in an amount that is anywhere between 1 / 10,000 and 1 / 2 the chemotherapeutic effective dose of the chemotherapeutic agent. For example, the chemotherapeutic agent may be administered in an amount that is about 1 / 20, about 1 / 500, or even about 1 / 5000 the chemotherapeutic effective dose of the chemotherapeutic agent. The chemotherapeutic agents of the present disclosure may be tested in vivo for the desired therapeutic activity in combination with the construct, as well as for the determination of effective dosages. For example, such compounds may be tested in suitable animal model systems, including but not limited to rats, mice, chickens, cows, monkeys, rabbits, and the like, prior to testing in humans. In vitro testing may also be used to determine suitable combinations and dosages, as described in the Examples.

[0084] G. Radiation Therapy In some embodiments, the cancer treatment of the present disclosure comprises radiation, such as ionizing radiation.As used herein, "ionizing radiation" refers to radiation, including particles or photons that have sufficient energy to cause ionization (gain or loss of electrons) or can generate sufficient energy through nuclear interaction.An exemplary and preferred ionizing radiation is x-ray.Means for delivering x-ray to target tissue or cell are well known in the art.

[0085] In some embodiments, the amount of ionizing radiation is greater than 20 Gy and is administered in one dose. In some embodiments, the amount of ionizing radiation is 18 Gy and is administered in three doses. In some embodiments, the amount of ionizing radiation is at least 2, 4, 6, 8, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 18, 19, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 40 Gy, but not more than 2, 4, 6, 8, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 2 ... 8, 19, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 40 Gy, or exactly 2, 4, 6, 8, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 18, 19, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 40 Gy (or any range derivable therein). In some embodiments, ionizing radiation is administered in at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 doses, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 doses, or exactly 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 doses (or any range derivable therein). If more than one dose is administered, the doses may be spaced apart by about 1, 4, 8, 12, or 24 hours, or 1, 2, 3, 4, 5, 6, 7, or 8 days, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, or 16 weeks, or any range derivable therein.

[0086] In some embodiments, the amount of IR can be referred to as a total dose of IR, which is then administered in fractionated doses. For example, in some embodiments, the total dose is 50 Gy administered in 10 fractionated doses of 5 Gy each. In some embodiments, the total dose is 50-90 Gy administered in 20-60 fractionated doses of 2-3 Gy each. In some embodiments, the total dose of IR is at least TIFF2024527109000032.tif41159, at most TIFF2024527109000033.tif41159, or approx. TIFF2024527109000034.tif41159 (or any range derivable therein). In some embodiments, the total dose is administered in fractional doses of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 20, 25, 30, 35, 40, 45, or 50 Gy, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 20, 25, 30, 35, 40, 45, or 50 Gy, or exactly 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 20, 25, 30, 35, 40, 45, or 50 Gy (or any range derivable therein). TIFF2024527109000035.tif 33159 times, at most TIFF2024527109000036.tif 33159 times or exactly In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 fractions, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 fractions, or exactly 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 fractions (or any range derivable therein) are administered per day. In some embodiments, the IL-15 antibody is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 times, but not more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 times. 2, 23, 24, 25, 26, 27, 28, 29, or 30 fractions, or exactly 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 fractions (or any range derivable therein) are administered per week.

[0087] H. Surgery Approximately 60% of people with cancer undergo some type of surgery, including preventive, diagnostic or staging, curative, and palliative surgery. Curative surgery includes resection, which physically removes, excises, and / or destroys all or part of the cancerous tissue, and may be combined with other treatments, such as the present embodiment of treatment, chemotherapy, radiation therapy, hormone therapy, gene therapy, immunotherapy, and / or alternative therapies. Tumor resection refers to the physical removal of at least a portion of the tumor. In addition to tumor resection, surgical treatments include laser surgery, cryosurgery, electrosurgery, and microsurgery (Mohs surgery).

[0088] Upon removal of part or all of cancerous cells, tissues, or tumors, a cavity may be formed in the body.Treatment may be achieved by perfusion, direct injection, or local application of additional anticancer treatment to the area.Such treatment may be repeated, for example, every 1, 2, 3, 4, 5, 6, or 7 days, or every 1, 2, 3, 4, and 5 weeks, or every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.Such treatment may also be of various dosages.

[0089] II. General Pharmaceutical Compositions In some embodiments, a pharmaceutical composition is administered to a subject. Various aspects may involve administering an effective amount of a composition to a subject. Such compositions are generally dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[0090] The phrase "pharmacologically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not cause adverse, allergic or other untoward reactions when administered to animals or humans.As used herein, "pharmacologically acceptable carriers" include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like.The use of such media and agents for pharmacoactive substances is well known in the art.Unless any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic composition is contemplated.Other supplementary active ingredients, such as anti-infective agents and vaccines, can also be included in the composition.

[0091] Active compound can be formulated for parenteral administration, for example, can be formulated for injection via intravenous, intramuscular, subcutaneous or intraperitoneal route.Typically, such compositions can be prepared as either liquid solution or suspension;solid forms can also be prepared that are suitable for use in preparing solution or suspension by adding liquid before injection;preparation can also be emulsified.

[0092] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations, for example, containing aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that it can be easily squirted. It must also be stable under the conditions of manufacture and storage, and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

[0093] The compounds of the present invention may form solvates, which are understood to be complexes of variable stoichiometry formed by a solute (e.g., poziotinib or its salt) and a solvent. Such solvents for the purposes of the present invention may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, dimethylsulfoxide, ethanol, and acetic acid. In some embodiments, the solvent is a pharma- ceutically acceptable solvent. In some embodiments, the solvent is water.

[0094] The pharmaceutical composition may contain a solvent or dispersion medium, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include an isotonic agent, for example, sugar or sodium chloride. Prolonged absorption of the injectable composition can be brought about by the use of absorption-delaying agents in the composition, for example, aluminum monostearate and gelatin.

[0095] Sterile injectable solution is prepared by incorporating the required amount of active compound into a suitable solvent with various other ingredients as listed above, as necessary, and then sterilizing by filtration or equivalent procedures.Generally, dispersion is prepared by incorporating various sterilized active ingredients into a sterile vehicle that contains a basic dispersion medium and other necessary ingredients from those listed above.In the case of sterile powder for preparing sterile injectable solution, the preferred method of preparation is vacuum drying and freeze-drying technique, which obtains a powder of active ingredient with any additional desired ingredients from its pre-sterilized filtered solution.

[0096] The administration of the composition is typically via any common route. This includes, but is not limited to, oral or intravenous administration. Alternatively, administration can be by orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal, or intranasal administration. Such compositions will usually be administered as a pharmaceutically acceptable composition that includes a physiologically acceptable carrier, buffer, or other excipient.

[0097] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutically or prophylactically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above.

[0098] III. Kits Certain aspects of the present disclosure also relate to kits comprising the compositions of the present disclosure or compositions for carrying out the methods of the present disclosure. In some embodiments, the kits can be used to evaluate one or more biomarkers. In certain embodiments, the kits can be used to evaluate one or more biomarkers. In certain embodiments, the kits can be used to evaluate one or more biomarkers, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 100, 500, 1,000 or more, or includes at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 100, 500, 1,000 or more, or any value or range and combination derivable therein, or at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 100, 500, 1,000 or more, or any values ​​or ranges and combinations derivable therein. In some embodiments, there are kits for assessing biomarker activity in a cell.

[0099] The kits may include components, which may be individually packaged or disposed in containers, such as tubes, bottles, vials, syringes, or other suitable container means.

[0100] Individual components may be provided in the kit in concentrated amounts; in some embodiments, a component is provided individually at the same concentration as it would be in solution with the other components. Concentrations of components may be provided as 1×, 2×, 5×, 10×, or 20×, or higher.

[0101] Kits for using the disclosed probes, synthetic nucleic acids, non-synthetic nucleic acids, and / or inhibitors for prognostic or diagnostic applications are included as part of this disclosure. In particular, kits for using any of the biomarkers identified herein (e.g., KRAS G12C ) are contemplated, including nucleic acid primers / primer sets and probes identical to or complementary to all or a portion of the biomarker, which may include non-coding sequences of the biomarker as well as coding sequences of the biomarker.

[0102] In certain aspects, negative and / or positive control nucleic acids, probes, and inhibitors are included in some kit embodiments. Additionally, the kits may include samples that are negative or positive controls for one or more biomarkers.

[0103] Any embodiment of the disclosure that includes a particular biomarker by name is intended to also encompass embodiments that include biomarkers whose sequence is at least 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% identical to the mature sequence of the particular nucleic acid. EXAMPLES

[0104] The following examples are included to demonstrate preferred embodiments of the invention. It will be understood by those of skill in the art that the procedures disclosed in the examples follow representative procedures discovered by the inventors to work well in the practice of the invention, and therefore can be considered to constitute preferred modes for its practice. However, those of skill in the art, in light of the present disclosure, will appreciate that many changes can be made to the specific embodiments disclosed without departing from the spirit and scope of the invention and still obtain a similar or similar result.

[0105] Example 1 - Poziotinib synergistically enhances the activity of KRAS G12C inhibition in models of KRAS mutant cancer KRAS by AMG 510 (Sotorasib) in KRAS mutant lung cancer cell line H23 G12C Inhibition of poziotinib resulted in upregulation of pEGFR, pErbB3, and pErbB4 after 4 hours (Figure 1A). Poziotinib alone had little effect on KRAS mutant cell line viability at concentrations below 10,000 nM, whereas the addition of 100 nM poziotinib significantly reduced KRAS expression in KRAS mutant NSCLC cell lines. G12C IC of inhibitors AMG 510 and MRTX849 50 In H358 cells, the IC values ​​of AMG 510 and MRTX849 were 50 The IC values ​​for AMG 510 and MRTX849 decreased from 8 nM and 28 nM to 0.20 nM and 0.19 nM, respectively, in H1373 cells. 50 The IC values ​​for AMG 510 and MRTX849 decreased from 1,420 nM and 632 nM to 15.0 nM and 22.9 nM, respectively, in H1792 cells. 50 The IC values ​​for AMG 510 and MRTX849 decreased from >10,000 nM and 1,330 nM to 453 nM and 16.4 nM, respectively. 50The values ​​decreased from >10,000 nM and 1,360 nM to 472 nM and 3.67 nM, respectively. These results are shown in Figure 1B (for AMG 510) and Figure 1C (for MRTX849).

[0106] Example 2 - KRAS G12C inhibitors increase EGFR / HER2 phosphorylation NSCLC cell lines harboring KRAS G12C mutations were treated with sotrasib or adagrasib for 4 hours. pEGFR and pHER2 levels were assessed by RPPA. KRAS G12C inhibitor treatment resulted in significantly increased levels of pEGFR and pHER2. These results are shown in Figure 2.

[0107] Example 3 – KRAS G12C inhibitor treatment increases phosphorylation of all EGFR / HER family members HCC44, H2122, and H358 NSCLC cells (all harboring KRAS G12C mutations) were treated with adagrasib or sotorasib for 72 hours. Phosphorylation of ERBB family members was assessed by ELISA assay. Results after adagrasib treatment are shown in Figure 3A. Results after sotorasib treatment are shown in Figure 3B. In all cell lines, KRAS G12C inhibitor treatment caused induction of pEGFR, pHER2, pHER3, and pHER4.

[0108] Example 4 - Poziotinib has activity against HER family members, including HER4 Ba / F3 cells expressing EGFR, EGFR / HER2, HER2 / HER3, HER2 / HER4, HER3 / HER4, and HER4 were generated and tested for sensitivity to poziotinib and afatinib to evaluate the activity of these TKIs against each receptor. Results after poziotinib treatment are shown in Figure 4A. Results after afatinib treatment are shown in Figure 4B. Poziotinib had strong activity against cells expressing HER2 / HER3, HER2 / HER4, HER3 / HER4, and HER4. In contrast, afatinib had no activity against cells expressing HER3 / HER4 and HER4.

[0109] Example 5 - Poziotinib is selective for HER2-4 over EGFR compared to other TKIs Figure 5 shows the IC of various TKIs, including erlotinib, gefitinib, tucatinib, TAS0728, afatinib, poziotinib, dacomitinib, neratinib, BDTX-189, mobocertinib, lazertinib, and osimertinib. 50 Values ​​are shown. Data were determined based on Ba / F3 cells expressing EGFR, EGFR / HER2, HER2 / HER3, HER2 / HER4, HER3 / HER4, and / or HER4 receptors, and values ​​are based on Ba / F3 wild-type EGFR IC 50 Normalized IC 50 value.

[0110] Example 6 - Exogenous EGF or NRG1 can promote resistance to KRAS G12C inhibitors H358 cells (KRAS G12C positive NSCLC) were treated with adagrasib or sotorasib alone or in combination with EGF to activate EGFR or NRG1 and activate other HER family members. Figure 6A shows the resistance of H358 cells to treatment with adagrasib in combination with exogenous EGF or NRG1. Figure 6B shows the resistance of H358 cells to treatment with sotorasib in combination with exogenous EGF or NRG1. EGF or NRG1 treatment reduced the sensitivity to adagrasib and sotorasib. These findings support the notion that activation of this pathway can promote resistance to KRAS G12C inhibitors.

[0111] Example 7 - Combination of G12C inhibitors with poziotinib is more synergistic than EGFR-specific inhibitors H23, HCC44, H2122, and H1792 cells (NSCLC carrying KRAS G12C mutation) were treated with sotrasib or adagrasib alone or in combination with afatinib or poziotinib. Figure 7A shows the synergistic effect of treating NSCLC cells carrying KRAS G12C mutation with sotrasib alone or in combination with poziotinib. Figure 7B shows the synergistic effect of treating NSCLC cells carrying KRAS G12C mutation with adagrasib alone or in combination with poziotinib. Poziotinib, acting as a pan-HER inhibitor and EGFR inhibitor, produced a greater synergistic effect than afatinib, which only inhibits EGFR / HER2.

[0112] Example 8 - Poziotinib blocks G12C inhibitor-induced HER phosphorylation to a greater extent than afatinib HCC44, H2122, and H358 cells (all NSCLC carrying KRAS G12C mutation) were treated with sotrasib or adagrasib alone or with poziotinib or afatinib. The phosphorylation of ERBB family members pEGFR, pHER2, pHER3, and pHER4 was evaluated by ELISA assay. Figure 8A shows the phosphorylation of ERBB family members in KRAS G12C mutation-bearing NSCLC cells treated with sotrasib alone or in combination with afatinib or poziotinib. Figure 8B shows the phosphorylation of ERBB family members in KRAS G12C mutation-bearing NSCLC cells treated with adagrasib alone or in combination with afatinib or poziotinib. KRAS G12C inhibitors induced phosphorylation of EGFR and HER family receptors. This effect was inhibited by the addition of poziotinib to a greater extent than by the addition of afatinib.

[0113] Example 9 - Low-dose poziotinib enhances the in vivo activity of KRAS G12C inhibitors Sotorasib was treated with sotorasib alone or in combination with poziotinib (pozi) or afatinib (afat) in PDX models of KRAS G12C mutant NSCLC. Figure 9A shows the effect of sotorasib treatment alone or in combination with poziotinib (pozi) or afatinib (afat) on tumor volume in PDX models of KRAS G12C mutant NSCLC. Figure 9B shows the effect of sotorasib treatment alone or in combination with poziotinib (pozi) or afatinib (afat) on progression-free survival in PDX models of KRAS G12C mutant NSCLC. The addition of poziotinib enhanced the antitumor activity of sotorasib and prolonged the survival of animals.

[0114] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of this disclosure.Although the compositions and methods of the present invention are described with respect to preferred embodiments, it will be apparent to those skilled in the art that modifications can be made to the methods described herein and in the steps or in the sequence of steps of the methods without departing from the concept, spirit and scope of the invention.More specifically, it will be apparent that certain agents that are both chemically and physiologically related can be substituted for the agents described herein while still achieving the same or similar results.All such similar substitutions and modifications that are apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

[0115] References The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are expressly incorporated herein by reference. TIFF2024527109000038.tif57160

Claims

1. A pharmaceutical for use in a method of treating a subject for KRAS mutant cancer, comprising an effective amount of a KRAS inhibitor, said method comprising a step of administering said KRAS inhibitor to said subject, and said pharmaceutical being used in combination with an effective amount of poziotinib.

2. A pharmaceutical comprising an effective amount of poziotinib for use in a method of treating a subject for KRAS mutant cancer, said method comprising administering said poziotinib to said subject, said pharmaceutical being used in combination with an effective amount of a KRAS inhibitor.

3. A pharmaceutical for use in a method of treating a subject for KRAS mutant cancer, comprising a combination of effective amounts of (a) a KRAS inhibitor and (b) poziotinib, said method comprising administering said (a) KRAS inhibitor and (b) poziotinib to said subject.

4. (a) The KRAS inhibitor and poziotinib are administered substantially simultaneously. (b) the KRAS inhibitor and poziotinib are administered sequentially; (c) a KRAS inhibitor is administered prior to administering poziotinib; or (d) the KRAS inhibitor is administered after poziotinib; The pharmaceutical composition according to any one of claims 1 to 3.

5. KRAS inhibitors inhibit KRAS G12C The pharmaceutical agent according to any one of claims 1 to 3, which is an inhibitor.

6. KRAS G12C The pharmaceutical composition of claim 5, wherein the inhibitor is sotorasib (AMG 510) or adagrasib (MRTX849).

7. A method comprising: G12C The pharmaceutical composition according to any one of claims 1 to 3, further comprising a step of detecting a mutation.

8. The method of any one of claims 1 to 3, wherein the subject has previously been treated with cancer therapy.

9. The pharmaceutical composition of claim 8, wherein the cancer treatment includes chemotherapy or a KRAS inhibitor.

10. The pharmaceutical composition of claim 8, wherein the subject has been determined to be resistant to cancer treatment.

11. The pharmaceutical composition according to any one of claims 1 to 3, wherein the KRAS mutant cancer is KRAS mutant non-small cell lung cancer, KRAS mutant colorectal cancer, or KRAS mutant pancreatic cancer.

12. The pharmaceutical composition according to any one of claims 1 to 3, wherein poziotinib is administered orally.

13. The medicament of any one of claims 1 to 3, wherein the KRAS inhibitor and poziotinib are administered once per day for multiple days, or twice per day for multiple days.

14. A pharmaceutical described in any one of claims 1 to 3, wherein the method further comprises a step of administering additional cancer treatment to the subject.

15. 15. The pharmaceutical of claim 14, wherein the additional cancer treatment comprises chemotherapy, radiation therapy, immunotherapy, or a combination thereof.

16. (a) KRAS inhibitors; (b) poziotinib; and (c) Pharmaceutically acceptable excipients 10. A pharmaceutical composition comprising:

17. KRAS inhibitors inhibit KRAS G12C 17. The pharmaceutical composition of claim 16, which is an inhibitor.

18. KRAS G12C 18. The pharmaceutical composition of claim 17, wherein the inhibitor is sotorasib (AMG 510) or adagrasib (MRTX849).

19. 17. The pharmaceutical composition of claim 16, wherein poziotinib is at a dose of 0.1 mg to 50 mg.

20. A pharmaceutical for use in a method of treating a subject for KRAS G12C non-small cell lung cancer, comprising an effective amount of a KRAS G12C inhibitor, said method comprising a step of administering said KRAS G12C inhibitor to said subject, said pharmaceutical being used in combination with an effective amount of poziotinib.

21. A pharmaceutical comprising an effective amount of poziotinib for use in a method of treating a subject for KRAS G12C non-small cell lung cancer, said method comprising administering said poziotinib to said subject, said pharmaceutical being used in combination with an effective amount of a KRAS G12C inhibitor.

22. An effective amount (a) KRAS G12C inhibitors; and (b) Poziotinib 1. A pharmaceutical composition for use in a method of treating a subject for KRAS G12C non-small cell lung cancer, comprising a combination of (a) a KRAS G12C inhibitor and (b) poziotinib, said method comprising administering to said subject (a) a KRAS G12C inhibitor and (b) poziotinib.

23. (a) KRAS G12C the inhibitor and poziotinib are administered substantially simultaneously; (b) KRAS G12C The inhibitor and poziotinib are administered sequentially. (c)KRAS G12C The inhibitor is administered prior to administering poziotinib, or (d) KRAS G12C The inhibitor is administered after administering poziotinib, The pharmaceutical composition according to any one of claims 20 to 22.

24. KRAS G12C The pharmaceutical agent according to any one of claims 20 to 22, wherein the inhibitor is sotorasib or adagrasib.