Combination Therapies for the Treatment of Cancer

Abstract

The present disclosure provides methods of treating cancer with combination therapy of an SHP2 inhibitor and an EGFR inhibitor.

Description

[Technical Field]

[0001] cross reference This application claims the interests of U.S. Provisional Patent Application No. 63 / 124,6657 filed on 11 December 2020, U.S. Provisional Patent Application No. 63 / 214,718 filed on 24 June 2021, U.S. Provisional Patent Application No. 63 / 253,012 filed on 6 October 2021, and U.S. Provisional Patent Application No. 63 / 277,561 filed on 9 November 2021, each of which is incorporated herein by reference in its entirety. [Background technology]

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

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

[0004] SHP2 plays an important role in basic cellular functions including proliferation, differentiation, cell cycle maintenance, and motility. By dephosphorylating its associated signaling molecules, SHP2 regulates multiple intracellular signaling pathways in response to a wide range of growth factors, cytokines, and hormones. The cellular signaling processes in which SHP2 is involved include the RAS-MAPK (mitogen-activated protein kinase), PI3K (phosphoinositide 3-kinase)-AKT, and JAK-STAT pathways.

[0005] SHP2 further plays a role in signal enhancement on this pathway and acts downstream of RTK and upstream of RAS. One common mechanism of resistance to pharmacological inhibition of MAPK signaling involves the activation of RTKs that promote the reactivation of MAPK signaling. RTK activation recruits SHP2 via direct binding and via adapter proteins. These interactions cause the conversion of SHP2 from a closed (inactive) conformation to an open (active) conformation. SHP2 is an important facilitator of RAS signaling reactivation and evades pharmacological inhibition in both primary and secondary resistance. Inhibition of SHP2 often achieves the effect of globally attenuating upstream RTK signaling that promotes oncogenic signaling and adaptive tumor escape (see Prahallad, A. et al. Cell Reports 12, 1978 - 1985 (2015); Chen YN, Nature 535, 148 - 152 (2016)), which incorporates by reference in its entirety all of its teachings, including but not limited to all methods, compounds, compositions, data, etc. for use with any of the embodiments and disclosures of this specification.

[0006] In addition to SHP2, the epidermal growth factor receptor (EGFR), a transmembrane protein that is a receptor for members of the epidermal growth factor family of extracellular protein ligands, also operates upstream of the RAS pathway. Opportunities to target signaling pathways from multiple angles and potentially improve the feedback loop upstream of Ras via SHP2 and EGFR provide opportunities to develop ways of using combination therapies. Described herein are various specific methods and compositions related to the use of SHP2 and EGFR inhibitors.

[0007] In a first aspect, the present disclosure provides a method of treating a subject having cancer, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula I:

[0008]

Chemical formula

[0009] In some embodiments, EGFR in the subject is constitutively expressed.

[0010] In some embodiments, cancer is characterized by EGFR gene copy number increase, EGFR gene amplification, 7-chromosome polysomatic mutations, EGFR L858R, EGFR exon 19 deletion / insertion (e.g., E746_A750del, E746_T751delinsI, E746_T751delinsIP, E746_S752delinsA, E746_S752delinsV, E746_S752delinsV, L747_S752del, L747_T751del, and L747_P753delinsS), EGFR L861Q, EGFR G719C, EGFR G719S, EGFR G719A, EGFR V765A, EGFR This includes EGFR mutations selected from T783A, EGFR exon 20 insertions (e.g., N771dup, N771_H773dup, and P772_H773dup), EGFR splice variants (e.g., Vii, Vvi, and Vii), EGFR A289D, EGFR A289T, EGFR A289V, EGFR G598A, EGFR G598V, EGFR T790M, and EGFR C797S.

[0011] In some embodiments, the cancer is lung cancer.

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

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

[0014] In some embodiments, the EGFR inhibitor is selected from osimertinib, dacomitinib, razertinib, nazartinib, neratinib, mobocertinib, afatinib, erlotinib, gefitinib, lapatinib, rifilafenib, amivantamab, cetuximab, panitumumab, necitumumab, mizotamab-kreztocrax, nimotuzumab, and vandetanib.

[0015] In some embodiments, the EGFR inhibitor is osimertinib.

[0016] In some embodiments, the EGFR inhibitor is erlotinib.

[0017] In some embodiments, the EGFR inhibitor is gefitinib.

[0018] In some embodiments, the EGFR inhibitor is lapatinib.

[0019] In some embodiments, the EGFR inhibitor is neratinib.

[0020] In some embodiments, the EGFR inhibitor is afatinib.

[0021] In some embodiments, the method includes the step of administering a third MAPK pathway inhibitor.

[0022] In some embodiments, administration is by oral infusion.

[0023] In some embodiments, the dosage of the compound of formula I ranges from 20 mg to 400 mg per day.

[0024] In some embodiments, the dose of the EGFR inhibitor of formula I ranges from 1 mg to 1500 mg per day.

[0025] In a second aspect, the Disclosure provides a method for treating a target lung cancer, wherein the method involves a therapeutically effective amount of Formula I:

[0026] [ka] This includes administering the compound or a pharmaceutically acceptable salt thereof in combination with osimertinib.

[0027] In some embodiments, the compound of formula I is administered once or twice daily.

[0028] In some embodiments, osimertinib is administered once or twice daily.

[0029] In some embodiments, the subject is human.

[0030] In some embodiments, the lung cancer is non-small cell lung cancer.

[0031] In some embodiments, lung cancer has an EGFR mutation.

[0032] In a third aspect, the disclosure provides a kit comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, and an EGFR inhibitor.

[0033] In some embodiments, the compound of formula I and the EGFR inhibitor are in separate packages.

[0034] In some embodiments, the kit further includes instructions for administering the contents of the kit to a subject for the treatment of cancer.

[0035] In some embodiments, the EGFR inhibitor is one or more of the following: osimertinib, dacomitinib, razertinib, nazartinib, neratinib, mobocertinib, afatinib, erlotinib, gefitinib, lapatinib, rifilafenib, amibantamab, cetuximab, panitumumab, necitumumab, mizotamab crezutoclax, nimotuzumab, and vandetanib.

[0036] In the final embodiment, the Disclosure provides a method for treating a target cancer, wherein the method applies Formula I:

[0037] [ka] The process includes administering, in combination with cetuximab, orally in a therapeutically effective dose, the compound or a pharmaceutically acceptable salt thereof.

[0038] In some embodiments, cetuximab is administered weekly.

[0039] In some embodiments, cetuximab is administered every other week.

[0040] In some embodiments, the cancer is squamous cell head and neck cancer (SCCHN).

[0041] In some embodiments, the cancer is pancreatic ductal adenocarcinoma (PDAC).

[0042] In some embodiments, the compound of formula I is administered once or twice daily.

[0043] In some embodiments, the subject is human.

[0044] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in doses between approximately 20 mg and approximately 260 mg per day.

[0045] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in doses between approximately 20 mg and approximately 60 mg per day.

[0046] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered in doses of about 40 mg per day or about 60 mg per day.

[0047] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily in doses between approximately 10 mg and approximately 100 mg.

[0048] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily in doses between approximately 20 mg and approximately 80 mg.

[0049] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as an oral composition.

[0050] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once or twice daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered twice daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered continuously over a 28-day cycle.

[0051] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in amounts ranging from approximately 10 mg to approximately 140 mg.

[0052] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a three-week cycle consisting of two weeks of compound administration followed by one week of compound non-administration.

[0053] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a four-week cycle including three weeks of compound administration followed by one week of compound non-administration.

[0054] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of 6 weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of 8 weeks.

[0055] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered three times a week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first, third, and fifth days of the week.

[0056] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered four times a week.

[0057] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a three-week cycle consisting of two weeks of compound administration followed by one week of compound non-administration.

[0058] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a 4-week cycle including 3 weeks of compound administration followed by 1 week of compound non-administration.

[0059] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice a week, on two days. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of eight weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered on the first and second days of each week.

[0060] In some embodiments, the cancer is selected from lung cancer, gastric cancer, liver cancer, colon cancer, kidney cancer, breast cancer, pancreatic cancer, pancreatic ductal adenocarcinoma (PDAC), juvenile myelomonocytic leukemia, neuroblastoma, melanoma, and acute myeloid leukemia. [Brief explanation of the drawing]

[0061] [Figure 1A] This shows data on the combination of compound I and the EGFR inhibitor osimertinib in the cell line HCC827. This data demonstrates that the combination of compound I and the EGFR inhibitor osimertinib shows a synergistic effect in vitro. [Figure 1B] Numerical data for the combination of compound I and the EGFR inhibitor osimertinib in the cell line NCI-H820 are shown. This data indicates that the combination of compound I and the EGFR inhibitor osimertinib exhibits a synergistic effect in vitro. [Figure 2A] The graphs show the percentage activity versus inhibitor concentration (logM) in CAL-27 cells treated with the compound of formula I alone (black circles, line 1) and in combination with cetuximab at 0.5 μg / ml (black squares, line 2), 1.0 μg / ml (black circles, line 3), or 2.5 μg / ml (black squares, line 4). [Figure 2B] The bar graph shows the percentage of CTG activity, indicating that cetuximab treatment alone reduced cell viability by approximately 40%. [Figure 3A] The graph shows the percentage activity versus inhibitor concentration (log M) in SCC-9 cells treated with the compound of formula I alone and in combination with cetuximab at 0.5 μg / ml (black square, line 2), 1.0 μg / ml (black circle, line 3), or 2.5 μg / ml (black square, line 4). [Figure 3B] The bar graph shows the percentage of CTG activity, indicating that cetuximab treatment alone reduced cell viability by approximately 15%. [Figure 4A] The graph shows the percentage activity versus inhibitor concentration (logM) in SCC-15 cells treated with the compound of formula I alone and in combination with cetuximab at 1.0 μg / ml (black square, line 2), 2.5 μg / ml (black circle, line 3), or 5.0 μg / ml (black square, line 4). [Figure 4B] The bar graph shows the percentage of CTG activity, indicating that cetuximab treatment alone reduced cell viability by less than 10%. [Figure 5A] The graph shows the percentage of activity versus inhibitor concentration (log M) in SCC-25 cells treated with the compound of formula I alone and in combination with cetuximab at 2.5 μg / ml (black square, line 2), 5.0 μg / ml (black circle, line 3), or 10.0 μg / ml (black square, line 4). [Figure 5B] The bar graph shows the percentage of CTG activity, indicating that cetuximab treatment alone did not reduce cell viability. [Figure 6A] This shows an immunoblot of approximately 50% inhibition of ERK1 / 2 phosphorylation activity between the compound of formula I and DMSO. [Figure 6B]The bar graph shows the quantified phosphorylated ERK1 / 2 bands, normalized by total ERK. Quantitative results showed that treatment with compound I alone or cetuximab alone reduced ERK1 / 2 phosphorylation by approximately 50% compared to DMSO-treated control cells, while the combination of compound I and cetuximab showed approximately 80% inhibition of ERK1 / 2 phosphorylation in the HPV-negative head and neck squamous cell carcinoma cell line CAL27. [Figure 7A] The table shows the Bliss synergy scores for the combination of compound I and osimertinib in the (erlotinib-resistant) HCC827 / ER1 cell line. [Figure 7B] The table shows the Bliss synergy scores for the combination of compound I and osimertinib in the (erlotinib-resistant) HCC827ER1 parent cell line. [Figure 8] The graphs show tumor volume over treatment periods for regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR L858R / T790M mutant NSCLC PDX model LUN2355-215. [Figure 9] The graphs show tumor volume over treatment periods with regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR delE746_E749 / T790M mutant and the MET-amplified NSCLC CDX model NCI-H820. [Figure 10] The graphs show tumor volume over treatment periods with regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR L858R mutant and ERBB2-overexpressing NSCLC PDX model LUN2005-143-9. [Figure 11] The graphs show tumor volume over treatment periods for regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR L858R mutant NSCLC PDX model LUN2005-234. [Figure 12]The graphs show tumor volume over treatment periods for regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR L858R / T790M mutant NSCLC PDX model LUN2355-128-33. [Figure 13] The graphs show tumor volume over treatment periods for regimens of formula I compound alone, osimertinib alone, and a combination of formula I compound and osimertinib in the EGFRex19del mutant erlotinib-resistant CDX model HCC827 / ER1(METamp)[E4957-U2101]. [Figure 14] The graphs show tumor volume over treatment periods in the RAS / RAF wild-type PDX model CRC049, with regimens of formula I compound alone, cetuximab alone, and a combination of formula I compound and cetuximab. [Figure 15] The graphs show tumor volume over treatment periods with regimens of compound I alone, cetuximab alone, and a combination of compound I and cetuximab in the RAS / RAF wild-type HPV-negative HNSCC CDX model FaDu. [Modes for carrying out the invention]

[0062] I. General This embodiment provides a method for treating a subject having cancer, wherein the subject is given formula I:

[0063] [ka] The procedure includes administering a therapeutically effective dose of the compound of formula I or a pharmaceutically acceptable salt thereof in combination with an EGFR inhibitor. The following examples demonstrate the unexpected synergistic effects of the combination. The combination therapies disclosed herein using the compound of formula I or a pharmaceutically acceptable salt thereof can show superior results compared to alternative combinations of SHP2 inhibitors used in combination with EGFR inhibitors. Furthermore, the combination of the SHP2 inhibitor of formula I and the EGFR inhibitor provides a method that allows for the use of lower doses of either agent used alone in monotherapy, which may help reduce potential side effects. In particular, the combination therapy may be effective in cancer cells having any EGFR mutation or overexpressing EGFR. Thus, such treatment is suitable for the use of companion diagnostics to assist in the selection of an appropriate patient population. These and other advantages will be recognized by those skilled in the art.

[0064] II. Definition Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art in the field covered by these embodiments. In addition, any method or material similar to or equivalent to those described herein may be used in the implementation of the embodiments herein. For the purposes of the embodiments disclosed herein, the following terms are defined:

[0065] As used herein, “a,” “an,” or “the” include not only embodiments having one member but also embodiments having multiple members. For example, the singular forms “a,” “an,” and “the” include multiple referents unless explicitly indicated otherwise in the context. Thus, for example, a reference to “a cell” includes multiple cells, and a reference to “the agent” includes one or more agents known to those skilled in the art.

[0066] "Pharmacologically acceptable excipients" refer to substances that assist in the administration and absorption of an active agent to a subject. Useful pharmaceutical excipients in this embodiment include, but are not limited to, binders, fillers, disintegrants, lubricants, surfactants, coatings, sweeteners, flavorings, and colorants. Those skilled in the art will recognize that other pharmaceutical excipients may be useful in this embodiment.

[0067] "To treat," "treating," and "treatment" refer to any objective or subjective parameters, including, for example, relief, remission, reduction or impairment of symptoms, improved patient tolerance to the condition or state, slowing or reduction of the rate of degeneration, decline toward the terminal stage of degeneration, and improvement of the patient's physical or mental health, as well as any signs of success in treating or improving an impairment, condition or state. Treatment or improvement of symptoms may be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric examination, and / or psychiatric evaluation.

[0068] "Administering" refers to oral administration, suppository administration, topical contact, parenteral administration, intravenous, intraperitoneal, intramuscular, intrafocal, intranasal or subcutaneous administration, intrathecal administration, or implantation of a sustained-release device to the subject, such as a mini osmotic pump. In the combination therapies disclosed herein, administration may occur at separate times, simultaneously, or substantially simultaneously.

[0069] As used herein, “concurrent administration” or “combined administration” means administering the compositions described herein simultaneously with, immediately before, or immediately after the administration of one or more additional therapies. The compounds provided herein may be administered alone or concurrently to a patient. Concurrent administration is intended to include concurrent or sequential administration of compounds, individually or in combination (one or more compounds). Concurrent administration is intended to include administration of compounds on the same day, within the same week, and / or within the same treatment schedule. Compounds may have different administration schedules, but may still be concurrently administered if they are administered within the same treatment schedule. For example, palbociclib may be administered once daily for three weeks within a four-week treatment schedule, and the compound of Formula I is concurrently administered with palbociclib if administered at any time within a four-week treatment schedule.

[0070] The “therapeutic dose” refers to the dose administered that produces a therapeutic effect. The exact dose depends on the purpose of the treatment and can be determined by those skilled in the art using known techniques (see, for example, Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins), each of which is incorporated herein by reference in whole with respect to all teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein. In sensitized cells, the therapeutic dose may often be lower than the conventional therapeutic dose for unsensitized cells.

[0071] "Inhibition," "blocking," and "inhibitor" refer to compounds that partially or completely block or prohibit a particular action or function, or methods of partially or completely blocking or prohibiting such action or function.

[0072] "Cancer" means all types of cancer, neoplasms, or malignant tumors found in mammals (e.g., humans), including, but not limited to, leukemia, lymphoma, carcinoma, and sarcoma. Exemplary cancers that can be treated with the compounds or methods provided herein include brain cancer, glioma, glioblastoma, neuroblastoma, prostate cancer, colorectal cancer, pancreatic cancer, medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, head cancer, Hodgkin's disease, and non-Hodgkin lymphoma. Exemplary cancers that can be treated with the compounds or methods provided herein include cancers of the thyroid, endocrine system, brain, breast, cervix, colon, head and neck, liver, kidney, lung, ovary, pancreas, rectum, stomach, and uterus. Further examples include thyroid cancer, cholangiocarcinoma, pancreatic adenocarcinoma, cutaneous melanoma, colon adenocarcinoma, rectal adenocarcinoma, gastric adenocarcinoma, esophageal cancer, head and neck squamous cell carcinoma, invasive breast cancer, lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung cancer, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocythemia, primary macroglobulinemia, primary brain tumor, malignant pancreatic insulanoma, malignant carcinoid, bladder cancer, pre-malignant cutaneous lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical carcinoma, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid carcinoma, medullary thyroid cancer, melanoma, colorectal cancer, papillary thyroid carcinoma, hepatocellular carcinoma, pancreatic ductal adenocarcinoma (PDAC), or prostate cancer.

[0073] "EGFR inhibitor" refers to any inhibitor of wild-type EGFR or any EGFR variant. EGFR variants include, but are not limited to, any of those disclosed in U.S. Patent Publication 2018 / 0235968, which is incorporated herein by reference in its entirety. EGFR variants include, but are not limited to, single nucleotide polymorphisms, exon insertions and deletions, and polysomatic variants. Specific examples of mutations include, but are not limited to, EGFR gene copy number increase, EGFR gene amplification, chromosome 7 polychromosome, EGFR L858R, EGFR exon 19 deletion / insertion (e.g., E746_A750del, E746_T751delinsI, E746_T751delinsIP, E746_S752delinsA, E746_S752delinsV, E746_S752delinsV, L747_S752del, L747_T751del, and L747_P753delinsS), EGFR L861Q, EGFR L718Q, EGFR G719C, EGFR G719S, EGFR G724S, EGFR G719A, EGFR V765A, EGFR This includes T783A, EGFR exon 20 insertions (e.g., N771dup, N771_H773dup, and P772_H773dup), EGFR splice variants (e.g., Vii, Vvi, and Vii), EGFR A289D, EGFR A289T, EGFR A289V, EGFRG G598A, EGFR G598V, EGFR S768I, EGFR T790M, and EGFR C797S. In some embodiments, one or more of the variants detailed in this paragraph and elsewhere in this specification can be specifically excluded from embodiments described herein, including but not limited to any methods, kits, and compositions, etc. Non-exclusive examples of EGFR inhibitors include osimertinib, dacomitinib, razertinib, nazartinib, neratinib, mobocertinib, afatinib, erlotinib, gefitinib, lapatinib, rifilafenib, amibantamab, cetuximab, panitumumab, necitumumab, mizotamab crezutoclax, nimotuzumab, and vandetanib.Other EGFR inhibitors include those disclosed in U.S. Patent Applications Publications 2020 / 0002279, 2019 / 0202920, and 2019 / 0167686, and International Applications WO2012 / 061299, WO2019 / 067543, and WO2020 / 190765, each of which is incorporated herein by reference in whole. In some embodiments, one or more inhibitors detailed in this paragraph and elsewhere in this specification, as well as inhibitors in incorporated references, may be specifically excluded from one or more embodiments described herein, including, but not limited to, any methods, kits, and compositions.

[0074] "Subject" refers to an organism that is suffering from or susceptible to a disease or illness that can be treated by administration of a pharmaceutical composition such as those provided herein. Examples, but not limited to, include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, deer, horses, and other non-mammals. In some embodiments, the patient is human.

[0075] III. Method of Administration In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is formulated as an oral composition.

[0076] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once or twice daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered twice daily. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered continuously over a 28-day cycle.

[0077] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in amounts ranging from approximately 10 mg to approximately 140 mg.

[0078] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a three-week cycle consisting of two weeks of compound administration followed by one week of compound non-administration.

[0079] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a four-week cycle including three weeks of compound administration followed by one week of compound non-administration.

[0080] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of 6 weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of 8 weeks.

[0081] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered three times a week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first, third, and fifth days of the week.

[0082] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered four times a week.

[0083] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a three-week cycle consisting of two weeks of compound administration followed by one week of compound non-administration.

[0084] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a 4-week cycle including 3 weeks of compound administration followed by 1 week of compound non-administration.

[0085] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily for two days a week. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of eight weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered on the first and second days of each week.

[0086] In some embodiments, the cancer is selected from lung cancer, gastric cancer, liver cancer, colon cancer, kidney cancer, breast cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, and acute myeloid leukemia. In some embodiments, the cancer is pancreatic ductal adenocarcinoma (PDAC).

[0087] IV. Combination Therapy In some embodiments, methods for treating subjects having cancer are provided herein, the methods for treating subjects with formula I:

[0088] [ka] The procedure includes administering a therapeutically effective dose of the compound or a pharmaceutically acceptable salt thereof in combination with an EGFR inhibitor. As disclosed herein, significant synergistic effects were observed beyond those expected for such combinations.

[0089] In some embodiments, the methods disclosed herein are suitable for the treatment of any cancer in which EGFR plays a role. In some embodiments, the cancer may be, for example, colorectal cancer (e.g., colon cancer, rectal cancer, etc.). In some embodiments, the cancer may be, for example, NSCLC (non-small cell lung cancer). In some embodiments, the cancer may be glioma, including, for example, glioblastoma. In some embodiments, the cancer is trinegative breast cancer. In some embodiments, the cancer is thyroid cancer. In some embodiments, the cancer may be, for example, head and neck squamous cell carcinoma. As will be understood by those skilled in the art, tumors can metastasize from the first or primary locus of the tumor to one or more other body tissues or sites. In particular, metastases to the central nervous system (i.e., secondary CNS tumors), and especially to the brain (i.e., brain metastases), are well documented for tumors and cancers, such as breast, lung, melanoma, kidney, and colorectal cancer. Thus, the methods disclosed herein can also be used for the treatment of metastases to other organs (i.e., metastatic tumor growth).

[0090] In some embodiments, the method may include the step of administering a third MAPK pathway inhibitor. Without being limited by theory, suppression of MAPK signaling in cancer cells may result in downregulation of PD-L1 expression, increasing the likelihood that cancer cells will be detected by the immune system. Such third MAPK pathway inhibitors may be based on other mutations in proteins in the MAPK pathway. In some embodiments, any MAPK pathway inhibitor can be used, including those targeting K-Ras, N-Ras, H-Ras, PDGFRA, PDGFRB, MET, FGFR, ALK, ROS1, TRKA, TRKB, TRKC, EGFR, IGFR1R, GRB2, SOS, ARAF, BRAF, RAF1, MEK1, MEK2, c-Myc, CDK4, CDK6, CDK2, ERK1, and ERK2. Exemplary MAPK pathway inhibitors include, but are not limited to, afatinib, osimertinib, erlotinib, gefitinib, lapatinib, neratinib, dacomitinib, vandetanib, cetuximab, panitumumab, nimotuzumab, necitumumab, trametinib, binimetinib, cobimetinib, selumetinib, urixertinib, LTT462, and LY3214996. In some embodiments, but are not limited, one or more of the above-described inhibitors, including any methods, kits, and compositions, may be specifically excluded from the embodiments described herein.

[0091] The methods disclosed herein can be combined with other chemotherapeutic agents. Examples of such anticancer agents can be found in *Cancer Principles and Practice of Oncology* by VTDevita and S. Hellman (editors), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers, which is incorporated herein by reference in its entirety, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein. Those skilled in the art will be able to identify which combinations of agents are useful based on the specific characteristics of the drugs and diseases involved.

[0092] In some embodiments, the method may involve the simultaneous administration of at least one cytotoxic agent. The term “cytotoxic agent” means, as used herein, a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., radioisotopes of At211, I131, I125, Y90, Rel86, Re188, Sm153, Bi212, P32, Pb212, and Lu), chemotherapeutic agents, growth inhibitors, enzymes such as nucleases and their fragments, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin, including fragments and / or variants thereof.

[0093] Examples of cytotoxic agents may be selected from antimicrotubule agents, platinum coordination complexes, alkylating agents, antibiotics, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signaling pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, apoptosis promoters, LDH-A inhibitors; fatty acid biosynthesis inhibitors; cell cycle signaling inhibitors; HDAC inhibitors; proteasome inhibitors; and cancer metabolism inhibitors.

[0094] Chemotherapy agents include compounds useful for treating cancer. Examples of chemotherapeutic agents include erlotinib (TARCEVA®, Genentech / OSI Pharm.), bortezomib (VELCADE®, Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX®, AstraZeneca), sunitinib (SUTENT®, Pfizer / Sugen), letrozole (FEMARA®, Novartis), and imatinib mesylate. Glucosin (GLEEVEC®, Novartis), finasunate (VATALANIB®, Novartis), oxaliplatin (ELOXATIN®, Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib (TYKERB®, GSK572016, Glaxo Smith Kline), lonafarnib (SCH 66336), sorafenib (NEXAVAR®, Bayer Labs), gefitinib (IRESSA®, AstraZeneca), AG1478, alkylating agents, e.g., thiotepa and cytoxane®. Cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan, and biposulfan; aziridines such as benzodopa, carbocone, meturedopa, and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine; acetogenins (especially bratacin and bratacinone);Camptothecin (containing topotecan and irinotecan); bryostatin; calistatin; CC 1065 (containing its adzelesin, karzelesin, and biseresin synthetic analogs); cryptophycin (especially cryptophycin 1 and cryptophycin 8); corticosteroids (containing prednisone and prednisolone); cyproterone acetate; 5-α-reductase containing finasteride and dutasteride; vorinostat, romidepsin, panobinostat, valproic acid, mosetinostat; aldesleukin, talc duocalmycin (containing synthetic analogs, kW-2189 and CB1-TM1)); eleutherobin; pancratistatin; sarcodictiin A (a sarcodictyin; spongistatin; chlorambutyl, chlornafadin, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembichin, phenesterine, prednimustine, trophosphamide, uracil mustard and other nitrogen mustards; nitrosourea such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; engineeric antibiotics (e.g., calicheamicin, especially calicheamicin γ1I and calicheamicin ω1I (Angew Chem. Intl. Ed. Engl. 1994) Antibiotics such as 33:183-186); dynemycin including dynemicin A; bisphosphonates such as clodronate; esperamicin;(and neocardinostatin chromophores and related pigment proteins enediin antibiotic chromophores), aclasinomycin, actinomycin, ausramycin, azaserin, bleomycin, kactinomycin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin such as mitomycin C, mycophenolic acid, nogaramycin; Olivomycin, peplomycin, porphyromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folate analogs such as denopterin, methotrexate, pteropterin, and trimethrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; ancitabine, azacitidine, 6 Pyrimidine analogs such as azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and fulocuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testactone; anti-adrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid replenishers such as frolinic acid; acegraton; and aldophosphamide glycoside. glycoside; aminolevulinic acid; enyluracil; amsacrine; bestrabusil; bisanthren; edatraxate; defofamine; demecoltin; diazicone; elfomithine; eriptinium acetate; epotilon; etogluside; gallium nitrate; hydroxyurea; lentinan; lonidainine; mytansinoids such as mytansin and anthamitosin; mitoxantrone; mopidamnol; nitraerine; pentostatin; fenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; PSKR polysaccharide complex (JHS Natural Products, Eugene, Oreg.)); Lazoxane; Rhizoxin; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadiquan; 2,2',2''-Trichlorotriethylamine; Trichothecenes (especially T-2 toxin, veraculin A, roridin A, and anguidine); Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"); Cyclophosphamide; Thiotepa; Taxoids, e.g., TAXOL (Paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), Abraxane (registered trademark) (excluding Cremofol), Albumin-modified nanoparticle formulations of Paclitaxel (American Pharmaceutical Partners, Schaumberg, III.), and Taxotere® (docetaxel; Sanofi-Aventis); chlorambucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE® (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO). This includes retinoids such as retinic acid; and pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0095] The chemotherapeutic agents also include (i) anti-hormone agents that act to modulate or inhibit the hormonal effects on tumors, such as anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (NOLVADEX®; including tamoxifen citrate), raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, and FARESTON® (toremifene citrate). (ii) Aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal gland, such as 4(5)-imidazole, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestany, fadrozol, RIVISOR® (borozol), FEMARA® (letrozole; Novartis), and ARIMIDEX® (anastrozole; AstraZeneca); (iii) Antiandrogens such as flutamide, nylutamide, bicalutamide, leuprolide, and goserelin; buserelin, trypterine, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all transretinic acids, fenretinide, and troxacitabine (1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those involved in signaling pathways associated with abnormal cell proliferation. (vii) Ribozymes such as VEGF expression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii) Vaccines such as gene therapy vaccines, e.g., ALLOVECTIN®, LEUVECTIN®, and VAXID®; PROLEUKIN®, rIL-2; topoisomerase 1 inhibitors such as LURTOTECAN®; ABARELIX® rmRH;and (ix) comprising any pharmaceutically acceptable salt, acid, and derivative thereof;

[0096] The chemotherapy agents further include antibodies and antibody-drug conjugates such as alemtuzumab (Campus), bevacizumab (AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); panitumumab (VECTIBIX®, Amgen); rituximab (RITUXAN®, Genentech / Biogen Idec); pertuzumab (OMNITARG®, 2C4, Genentech); trastuzumab (HERCEPTIN®, Genentech); and tositumomab (Bexxar, Corixia), as well as gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Further humanized monoclonal antibodies that have therapeutic potential as drugs when combined with the compounds of the present invention include apolizumab, aselizumab, atlizumab, bapineuzumab, vivacuzumab meltansine, cantuzumab meltansine, sedelizumab, certolizumab pegol, sidofcituzumab, sidotuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erulizumab, felbizumab, fontrizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, rabetuzumab, lintuzumab, matsuzumab, mepolizumab, motabizumab, motobizumab, and nata Lizumab, nimotuzumab, norovizumab, numabizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecufcituzumab, pectuzumab, pexerizumab, larivizumab, ranibizumab, reslivizumab, reslizumab, resivizumab, roberizumab, luprizumab, cibrotuzumab, cipri This includes zumab, sontuzumab, tacutuzumab tetraxetan, tadocituzumab, tarizumab, tefivazumab, tocilizumab, tralizumab, tucotsuzumab cermoloykin, tucucituzumab, umabituzumab, urtoxazumab, ustekinumab, bicilizumab, and anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), which is a recombinant, exclusively human-sequence full-length IgG1λ antibody genetically modified to recognize the interleukin-12 p40 protein.

[0097] Chemotherapy agents further include other “EGFR inhibitors,” which refer to compounds that bind to EGFR or its variant forms, or otherwise directly interact with it, thereby preventing or reducing its signaling activity, and are substituted for “EGFR antagonists.” Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB 8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al.) and their variants, e.g., chimeric 225 (C225 or cetuximab; ERBUTIX) and remodeled human 225 (H225) (WO96 / 40210, Imclone Systems). See Inc.); IMC-11F8 (Imclone), a fully human EGFR-targeting antibody; antibodies that bind to type II mutant EGFR (U.S. Patent No. 5,212,290); humanized and chimeric antibodies that bind to EGFR as described in U.S. Patent No. 5,891,996; human antibodies that bind to EGFR such as ABX-EGF or panitumumab (see WO98 / 50433, Abgenix / Amgen); EMD 55900 (Stragliotto et al. Eur.J. Cancer) 32A:636-640(1996)) Humanized EGFR antibodies against EGFR that compete with both EGF and TGFα for EGFR binding include EMD7200 (matsuzumab) (EMD / Merck); a fully human antibody known as the human EGFR antibody, HuMax-EGFR (GenMab); E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 and E7.6.3, and described in US6,235,883; MDX-447 (Medarex Inc); and mAb806 or humanized mAb806 (Johns et al., J. Biol. Chem. 279(29):30375-30384(2004)).Anti-EGFR antibodies can be conjugated with cytotoxic agents to generate immune complexes (see, for example, EP659,439A2, Merck Patent GmbH). EGFR antagonists are listed in U.S. Patents 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, and 5,866. This includes small molecules such as compounds described in PCT publications WO98 / 14451, WO98 / 50038, WO99 / 09016 and WO99 / 24037, as well as publications 572, 6399, 602, 602, 603, 6391, 674, 6344, 455, 5760, 041, 6002, 008 and 5747, 498.Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA-Genentech / OSI Pharmaceuticals); PD 183805 (CI 1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-182(N8-(3-chloro-4-fluorophenyl)-N2-(1-methyl-piperidine-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidine-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785(N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butinamide); EKB-569(N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide)(Wyeth); AG1478(Pfizer); AG1571(SU 5271; Pfizer); Dual EGFR / HER2 tyrosine kinase inhibitors, such as lapatinib (TYKERB®, GSK572016, or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine]). Each of the above references is incorporated herein by reference in its entirety, with respect to all teachings, including but not limited to all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein.

[0098] Chemotherapy agents also include "tyrosine kinase inhibitors," including EGFR-targeted drugs mentioned in the paragraph above; small molecule HER2 tyrosine kinase inhibitors such as TAK165 available from Takeda; oral selective inhibitors of ErbB2 receptor tyrosine kinase, such as CP-724, 714 (Pfizer and OSI); dual-HER inhibitors such as EKB-569 (available from Wyeth), which preferentially binds to EGFR but inhibits both HER2 and EGFR overexpressing cells; lapatinib (GSK572016; available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033; Pharmacia); and ISIS, which inhibits Raf-1 signaling. Raf-1 inhibitors such as the antisense agent ISIS-5132 available from Pharmaceuticals; non-HER-targeted TK inhibitors, e.g., imatinib mesylate (GLEEVEC®, available from Glaxo SmithKline); multi-targeted tyrosine kinase inhibitors, e.g., sunitinib (SUTENT®, available from Pfizer); VEGF receptor tyrosine kinase inhibitors such as batalanib (PTK787 / ZK222584, available from Novartis / Schering AG); MAPK extracellular regulatory kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines, e.g., PD153035,4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyridopyrimidines; pyrrolopyrimidines, e.g., CGP59326, CGP60261 and CGP62706; pyrazolopyrimidines, 4-(phenylamino)-7H-py Lolo[2,3-d]pyrimidine; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tilphostine containing the nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those that bind to nucleic acids encoding HER); quinoxaline (US Patent No. 5,804,396); triphostine (US Patent No. 5,804,396); ZD6474 (Astra Zeneca);PTK-787 (Novartis / Schering); pan-HER inhibitors, e.g., CI-1033 (Pfizer); Affinitac (ISIS 3521; ISIS / Lilly); Imatinib mesylate (GLEEVEC®); PKI166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); INC-1C11 (Imclone); Rapamycin (Sirolimus, RAPAMUNE®); or the following patent publications: U.S. Patent No. 5,804,396; WO1999 / 09016 (American Cyanamid); WO1998 / 43960 (American Cyanamid); WO1997 / 38983 (Warner Lambert); WO1999 / 06378 (Warner Lambert); WO1999 / 06396 (Warner Lambert); WO1996 / 30347 (Pfizer, Inc); WO1996 / 33978 (Zeneca); including those described in any of WO1996 / 3397 (Zeneca) and WO1996 / 33980 (Zeneca). Each of the above references is incorporated herein by reference in its entirety, with respect to all teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein.

[0099] The chemotherapy agents include dexamethasone, interferon, colchicine, methoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostin, arsenic trioxide, asparaginase, BCG Live, bevacuzimab, besarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexazoxane, epoetin alfa, erotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, and interferon alfa-2 b. Further comprising lenalidomide, levamisol, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelbequin, parifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexeddisodium, plicamycin, porfimer sodium, quinacrine, rasburicase, salglamostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, barulurubicin, zoledronate, and zoledronic acid, as well as pharmaceutically acceptable salts thereof.

[0100] The chemotherapy agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, thixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortone, and hydrocortisone-17-butyrate. Hydrocortisone-17-valerate, acromethasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetazone-17-butyrate, clobetasol-17-propionate, fluocortone caproate, fluocortone pivalate and flupredniden acetate; phenylalanine-glutamine-glycine (FEG) and its D-isomer (FEG) (IMULAN Immunoselective anti-inflammatory peptides (ImSAIDs), such as those used by BioTherapeutics, LLC; anti-rheumatic drugs, e.g., azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salt, hydroxychloroquine, leflunomideminocycline, sulfasalazine; tumor necrosis factor α (TNFα) blockers, e.g., etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), golimumab (Simponi), interleukin-1 (IL-1). Blockers, e.g., anakinra (Kineret); T-cell co-stimulation blockers, e.g., abatacept (Orencia); interleukin-6 (IL-6) blockers, e.g., tocilizumab (ACTEMERA®); interleukin-13 (IL-13) blockers such as lebrikizumab; interferon-alpha (IFN) blockers such as lontalizumab; beta-7 integrin blockers such as rhuMAb beta-7; IgE pathway blockers such as anti-M1 prime; homotrimer LTa3 and membrane-bound heterotrimer LTa / p2 blockers such as anti-lymphotoxin α (LTa); radioisotopes (e.g., At 211 , I 131 , I125 、Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 、P 32 、Pb 212and radioactive isotopes of Lu); other investigational drugs such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3, and farnesyltransferase inhibitors (L-739749, L-744832); polyphenols and their derivatives such as quercetin, resveratrol, piceatannol, epigallocatechin gallate, theaflavins, flavanols, procyanidins, and betulinic acid; autophagy inhibitors such as chloroquine; δ-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapaco Lapachol; Colchicine; Betulinic acid; Acetylcamptothecin, Scopolectin, and 9-Aminocamptothecin; Podophyllotoxin; Tegafur (UFTORAL®); Bexarotene (TARGRETIN®); Clodronate (e.g., BONEFOS® or OSTAC®), Etidronate (DIDROCAL®), NE-58095, Zoledronic acid / Zoledrone (ZOMETA®), Alendronate (FOSAMAX®), Bisphosphonates such as pamidronate (AREDIA®), tildronate (SKELID®), or risedronate (ACTONEL®); and epidermal growth factor receptor (EGF-R); vaccines such as THERATOPE® vaccine; perifosin, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); CCI-779; tipifarnib (R11577); olafenib, ABT510; oblimersen sodium (GENASENSE( This includes combinations of two or more of the above, such as Bcl-2 inhibitors (registered trademark)); pixantrone; farnesyltransferase inhibitors such as ronafarnib (SCH6636, SARASAR®); and CHOP, an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, along with pharmaceutically acceptable salts, acids, or derivatives of any of the above; and FOLFOX, an abbreviation for treatment regimens with oxaliplatin (ELOXATIN®) in combination with 5-FU and leucovorin.

[0101] Chemotherapy agents further include nonsteroidal anti-inflammatory drugs having analgesic, antipyretic, and anti-inflammatory effects. NSAIDs include non-selective inhibitors of the enzyme cyclooxygenase. Specific examples of NSAIDs include propionic acid derivatives such as aspirin, ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, and naproxen; acetic acid derivatives such as indomethacin, sulindac, etodolac, and diclofenac; enolic acid derivatives such as piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, and isoxicam; fenamic acid derivatives such as mefenamic acid, meclofenamic acid, flufenamic acid, and tolfenamic acid; and COX-2 inhibitors such as celecoxib, etoricoxib, lumiracoxib, parecoxib, lofecoxib, lofecoxib, and valdecoxib. NSAIDs may be indicated for the relief of symptoms of diseases such as rheumatoid arthritis, osteoarthritis, inflammatory arthritis, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headaches and migraines, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, ileus, and colitis.

[0102] In some embodiments, the chemotherapeutic agents include, but are not limited to, doxorubicin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, interferon, platinum derivatives, taxanes (e.g., paclitaxel, docetaxel), vinca alkaloids (e.g., vinblastine), anthracyclines (e.g., doxorubicin), epipodophilotoxin (e.g., etoposide), cisplatin, mTOR inhibitors (e.g., rapamycin), methotrexate, actinomycin D, dorastatin 10, colchicine, trimethrexate, methoprine, cyclosporine, daunorubicin, teniposide, amphotericin, alkylating agents (e.g., chlorambucil), 5-fluorouracil, camptothecin, cisplatin, metronidazole, and imatinib mesylate. In other embodiments, the compounds disclosed herein are administered in combination with a biological agent such as bevacizumab or panitumumab.

[0103] In certain embodiments, the compounds disclosed herein, or pharmaceutically acceptable compositions thereof, include avalerix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amiphostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG Live, bevacuzimab, fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, carsterone, capecitabine, camptothecin, carboplatin, carmustine, cetuximab, chlorambucil, cladribine, clofarabine, cyclophosphamide, cytarabine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin, dexazoxane, docetaxel, and doxorubicin.(Neutral), Doxorubicin hydrochloride, Dromostanolone propionate, Epirubicin, Epoetin alfa, Erotinib, Estramustine, Etoposide phosphate, Etoposide, Exemestane, Filgrastim, Floxuridine, Fludarabine, Fulvestrant, Gefitinib, Gemcitabine, Gemtuzumab, Goserelin acetate, Histrelin acetate, Hydroxyurea, Ibritumomab, Idarubicin, Ifosfamide, Me Imatinib silate, interferon α-2a, interferon α-2b, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisol, lomustine, megestrol acetate, melphalan, mercaptopurine, 6-MP, mesna, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone, nelarabine, nofetumomab, oprelbequine, oxalipin Latin, paclitaxel, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobromane, plicamycin, porfimer sodium, procarbazine, quinacrine, rasburicase, rituximab, salglamostim, sorafenib, streptozocin, sunitinib maleate, talc, tamoxifen, te It is administered in combination with one or more antiproliferative or chemotherapeutic agents selected from mozolomide, teniposide, VM-26, testactone, thioguanine, 6-TG, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, ATRA, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, zoledronate, or zoledronic acid.

[0104] In some embodiments, the administration of the compound of formula I may be any appropriate amount to treat cancer. For example, the administration may be a daily dose between 1 mg body weight and 500 mg. As a further example, the daily dose may be in the range of about 20 mg to 400 mg (or any partial range or partial value including the endpoints therein). In some embodiments, the administration range of the compound of formula I may be 10 mg to 300 mg. In some embodiments, the administration range of the compound of formula I may be 10 mg to 100 mg. In some embodiments, the administration range of the compound of formula I may be 5 mg to 50 mg. The daily dose can be achieved by administering a single dose (e.g., QD) or by administering multiple doses during the day (e.g., BID, TID, QID, etc.) to provide the total daily dose. In some embodiments, the administration of the EGFR inhibitor is any appropriate amount. For example, it may be an amount in the range of 1 mg to 500 mg per day (or any partial range or partial value including the endpoints therein). The dosage of an EGFR inhibitor may be the same as or less than the approved dosage for any given EGFR inhibitor and may depend on the given indication. For example, osimertinib for NSCLC may be administered at a daily dose of 20–100 mg in marketed doses of 40 mg and 80 mg. It will be understood that each of the detailed ranges described above may include any subrange or subpoint within it, including the endpoint. It will be understood that each of the detailed ranges described above may include any partial range or partialpoint within it, including the endpoint. The general dose range for adults is generally 5 mg to 2 g / day. Tablets or other forms of presentation provided in separate units may conveniently contain amounts of one or more compounds that are effective as a unit containing 5 mg to 500 mg, or a number of similar amounts, e.g., usually up to about 10 mg to 200 mg. The amount of active ingredient that can be combined with a carrier to produce a single dosage form varies depending on the host being treated and the specific mode of administration. In some embodiments, administration is oral. For example, cetuximab for squamous cell head and neck cancer (SCCHN) is administered by intravenous infusion at a dose of 500-100 mg / m². 2It can be administered in 50 ml and 200 mg / 100 ml vials, and commercially available doses are available. It will be understood that each of the detailed ranges described above may include any sub-range or subpoint within it, including the endpoint. It will be understood that each of the detailed ranges described above may include any sub-range or subpoint within it, including the endpoint. The general dose range for adult humans is generally 400 mg / m² for the first infusion. 2 From 250 mg / m² 2 Up to one infusion per week. Partial range is 500 mg / m². 2 / Every other week and 500 mg / m² 2 Includes bi-weekly infusions. Partial range is 250 mg / m². 2 / week, 250mg / m 2 / Every other week and 250 mg / m 2 Includes injections every 3 weeks. Partial range is 200 mg / m². 2 / week, 200mg / m 2 / Every other week and 200 mg / m 2 Includes infusions every 3 weeks. Partial range is 150 mg / m². 2 / week, 150mg / m 2 / Every other week and 150 mg / m 2 Includes injections every 3 weeks. Partial range is 100 mg / m². 2 / week, 100mg / m 2 / Every other week and 100 mg / m 2 Includes infusions every 3 weeks. Partial range is 500 mg / m². 2 / 120min, 400mg / m 2 / 120min, 250mg / m 2 / 60min, 250mg / m 2 / 120min, 200mg / m 2 / 60 minutes, 200mg / m 2 / 120min, 150mg / m 2 / 60min, 150mg / m 2 / 120min, 100mg / m 2 / 60 minutes, or 100 mg / m² 2This includes 120 minutes. In some embodiments, cetuximab may be administered for up to 54 months. In some embodiments, administration is by intravenous injection.

[0105] In some embodiments, a method is provided for treating colorectal cancer in a subject, comprising the step of orally administering a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof to the subject in combination with an EGFR inhibitor such as osimertinib. In some embodiments, the compound of formula I is administered once or twice daily. In some embodiments, osimertinib is administered once or twice daily. For example, drugs can be administered co-administered as described herein.

[0106] In some embodiments, a method for treating head and neck cancer (SCCHN) in a subject is provided, comprising the step of orally administering a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof to the subject in combination with an EGFR inhibitor such as cetuximab. In some embodiments, the compound of formula I is administered once or twice daily. In some embodiments, cetuximab is administered weekly. The drugs may be administered concurrently, for example, as described herein.

[0107] In some embodiments, patients with EGFR mutations (cancers with EGFR mutations) are selected.

[0108] In some embodiments, patients with NSCLC are selected. In some embodiments, patients with NSCLC with an EGFR mutation are selected. In some embodiments, patients with head and neck squamous cell carcinoma are selected. In some embodiments, patients with head and neck squamous cell carcinoma with an EGFR mutation are selected. In some embodiments, the stages of head and neck squamous cell carcinoma are classified stepwise. In some embodiments, Stage 0 generally refers to a tumor localized to the area where the tumor originated, and cancer cells are not present in deeper layers of tissue, nearby structures, lymph nodes, or distant sites. In some embodiments, Stage 1 generally refers to a primary tumor that is 2 cm or smaller and cancer cells are not present in deeper layers of tissue, nearby structures, lymph nodes, or distant sites. In some embodiments, Stage 2 generally refers to a tumor that measures 2-4 cm and cancer cells are not present in deeper layers of tissue, nearby structures, lymph nodes, or distant sites. In some embodiments, stage 3 generally refers to cases where the tumor is classified by one of the following criteria: (i) larger than 4 cm transversely, and cancer cells are not present in deeper layers of tissue, nearby structures, lymph nodes, or distal sites; or (ii) the tumor is of any size but has not proliferated into nearby structures or distal sites; cancer cells are present in one lymph node, which is located on the same head or neck side as the primary tumor and is less than 3 cm transversely. In some embodiments, head and neck squamous cell carcinoma is stage 2. In some embodiments, head and neck squamous cell carcinoma is stage 2 / 3. In some embodiments, head and neck squamous cell carcinoma is stage 3. In some embodiments, patients with colorectal cancer are selected. In some embodiments, patients with colorectal cancer with an EGFR mutation are selected.

[0109] In some embodiments, the cancer is negative for human papillomavirus (HPV). In some embodiments, the cancer lacks the KRAS mutation (wtKRAS). In some embodiments, the cancer lacks the NRAS mutation (wtNRAS). In some embodiments, the cancer lacks the BRAF mutation (wtBRAF). In some embodiments, the cancer is wtKRAS / wtNRAS / wtBRAF. In some embodiments, the cancer lacks mutations in KRAS, NRAS, or BRAF.

[0110] In some embodiments, the cancer has one or more acquired mutations. In some embodiments, the acquired mutations arise from the first-line treatment. In some embodiments, the first-line treatment is an EGFR inhibitor. In some embodiments, the EGFR inhibitor is osimertinib. In some embodiments, the EGFR inhibitor is cetuximab. In some embodiments, the cancer is a solid tumor carcinoma. In some embodiments, the cancer is NSCLC.

[0111] In some embodiments, the acquired mutation is an acquired EGFR mutation. In some embodiments, the acquired EGFR mutation is C797X. In some embodiments, the acquired EGFR mutation is L718Q. In some embodiments, the acquired EGFR mutation is EGFR amplification. In some embodiments, the acquired EGFR mutation is G724S. In some embodiments, the acquired mutation is S768I.

[0112] In some embodiments, the acquired mutation is an acquired amplification mutation. In some embodiments, the acquired mutation is MET gene amplification. In some embodiments, the acquired mutation is HER2 gene amplification.

[0113] In some embodiments, the acquired mutation is an acquired oncogenic fusion. In some embodiments, the acquired oncogenic fusion is SPTBN1-ALK. In some embodiments, the acquired oncogenic fusion is a RET fusion. In some embodiments, the acquired oncogenic fusion is a BRAF fusion.

[0114] In some embodiments, the acquired mutation is an acquired MAPK-PI3K mutation. In some embodiments, the acquired MAPK-PI3K mutation is BRAF-V600E. In some embodiments, the acquired MAPK-PI3K mutation is PI3KCA. In some embodiments, the acquired MAPK-PI3K mutation is KRAS. In some embodiments, the acquired MAPK-PI3K mutation is HER2.

[0115] In some embodiments, the subject is human. In some embodiments, the subject is a non-human mammal such as a primate, rodent, dog, cat, or other small animal.

[0116] composition The compounds of formula I disclosed herein may exist as salts. This embodiment includes such salts that may be pharmaceutically acceptable. Examples of applicable salt forms include salts with hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartaric acid (e.g., (+)-tartaric acid, (-)-tartaric acid, or mixtures thereof including racemic mixtures), succinate, benzoate, and amino acids such as glutamic acid. Such salts may be prepared by methods known to those skilled in the art. Salts with sodium, potassium, calcium, ammonium, organic amino, or magnesium, or similar salts are also included. If the compounds of this embodiment have relatively basic functionalities, acid addition salts may be obtained by contacting the neutral form of such compound with a sufficient amount of the desired acid in a neat or suitable inert solvent. Examples of acceptable acid addition salts This includes salts 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, as well as salts derived from organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, corticic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids such as alginates and salts of organic acids such as glucuronic acid or galactunoric acids are further included. Certain specific compounds in this embodiment possess both basic and acidic functionalities, allowing the compound to be converted into either a base or an acid addition salt.

[0117] Other salts include acid or base salts of the compounds used in the methods of this embodiment. Exemplary examples of pharmaceutically acceptable salts are mineral acid salts (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid), organic acid salts (e.g., acetic acid, propionic acid, glutamic acid, citrate), and quaternary ammonium salts (e.g., methyl iodide, ethyl iodide). It is understood that pharmaceutically acceptable salts are non-toxic. Further information regarding suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, which incorporates, by reference, all of its teachings, including but not limited to all methods, compounds, compositions, data, etc., for use with any of the embodiments and disclosures herein.

[0118] pharmaceutically acceptable salts include salts of the active compound prepared with a relatively non-toxic acid or base, depending on the specific substituents found in the compounds described herein. If the compounds of this embodiment include relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired base in a neat or suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts. If the compounds of this embodiment include relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired acid in a neat or suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, corticic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids such as alginates, and salts of organic acids such as glucuronic acid or galacturonic acid (see, for example, Berge et al, “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19) are also included, and all of their teachings, including but not limited to all methods, compounds, compositions, data, etc., for use with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety. Certain specific compounds of these embodiments possess both basic and acidic functionalities, which allow the compounds to be converted into either a base or an acid addition salt.

[0119] The neutral form of the compound is preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0120] Certain compounds of this embodiment can exist in solvated forms as well as non-solvated forms, including hydrated forms. Typically, the solvated forms are equivalent to the non-solvated forms and are encompassed within the scope of this embodiment. Certain compounds of this embodiment can exist in multiple crystalline or amorphous forms. Typically, all physical forms are intended to be suitable for the applications envisioned by this embodiment and are intended to be within the scope of this embodiment.

[0121] Certain compounds of this embodiment have an asymmetric carbon atom (optical center) or a double bond; the forms of enantiomers, racemates, diastereomers, tautomers, geometric isomers, and stereoisomers may be defined in terms of absolute stereochemistry as (R)- or (S)-, or (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of this embodiment. The compounds of this embodiment do not include those known to be too unstable in the art to be synthesized and / or separated. This embodiment is intended to include compounds in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or separated using conventional techniques.

[0122] Unless otherwise specified, the compounds of this embodiment may contain isotopes of atoms in an unnatural ratio in one or more of the atoms constituting such a compound. For example, the compounds of this embodiment may contain radioactive or stable isotopes, such as deuterium ( 2 H), tritium ( 3 H), Iodine-125( 125 I), Fluorine-18( 18 F), Nitrogen-15( 15 N), oxygen-17( 17 O), oxygen-18( 18O), carbon-13 ( 13 It can be labeled with C), or carbon-14 ( 14 C). All isotopic variations of the compounds of this embodiment, whether radioactive or not, are included within the scope of this embodiment.

[0123] In addition to salt forms, this embodiment provides compounds in prodrug form. The prodrugs of the compounds described herein are compounds that readily undergo chemical changes under the physiological conditions for providing the compounds of this embodiment. In addition, the prodrugs can be converted to the compounds of this embodiment by chemical or biochemical methods in an ex vivo environment. For example, the prodrug can be slowly converted to the compounds of this embodiment when placed in a transdermal patch reservoir with appropriate enzymes or chemical reagents.

[0124] In some embodiments, a pharmaceutical composition comprising a compound of formula I and a pharmaceutically acceptable excipient is provided. In some embodiments, the pharmaceutical composition is configured as an oral tablet preparation.

[0125] The compounds of this embodiment can be prepared and administered in a wide 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 the patient. The compounds of this embodiment can also be administered by injection (which is intravenous, intramuscular, intradermal, subcutaneous, intraduodenal, or intraperitoneal). They can also be administered by injection, i.e., intravenously, intramuscularly, intradermally, subcutaneously, intradurally, or intraperitoneally. Furthermore, the compounds described herein can be administered by inhalation, for example, intranasally. Furthermore, the compounds of this embodiment can be administered percutaneously. The compounds of formula I disclosed herein may also be administered by intraocular, intravaginal, and intrarectal routes, including suppositories, inhalants, powders, and aerosol formulations (for example, see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111, 1995), and all teachings, including but not limited to all methods, compounds, compositions, data, etc., for use with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety. Accordingly, these embodiments further provide pharmaceutical compositions comprising one or more pharmaceutically acceptable carriers and / or excipients and compounds of formula I or pharmaceutically acceptable salts of compounds of formula I.

[0126] For preparing pharmaceutical compositions from the compounds of this embodiment, the pharmaceutically acceptable carrier may be either solid or liquid. Solid preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. The solid carrier may be one or more substances that also act as diluents, fragrances, surfactants, binders, preservatives, tablet disintegrants, or encapsulating materials. Further details regarding the techniques for formulation and administration are described in detail in the scientific literature and patent documents, for example, the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's"), which, with reference to all teachings therein, including but not limited to all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein, are incorporated herein by reference in their entirety.

[0127] In dispersal, the carrier is a pulverized solid that is a mixture with the pulverized active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary binding properties in the appropriate ratio and additional excipients as needed, and then compressed into the desired shape and size.

[0128] Powders, capsules, and tablets preferably contain 5% or 10% to 70% of the active compound. Suitable carriers include magnesium carbonate, magnesium stearate, talc, sugars, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low-melting-point waxes, and cocoa butter. The term “preparation” is intended to include formulations of the active compound comprising an encapsulating material as a carrier providing a capsule, where the active ingredient, with or without other excipients, is surrounded by the carrier and thus attached to the carrier. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0129] Suitable solid excipients include sugars such as lactose, sucrose, mannitol, or sorbitol; starches from corn, wheat, rice, potato, or other plants; celluloses such as methylcellulose, hydroxypropylmethylcellulose, or sodium carboxymethylcellulose; and gums such as gum arabic and gum tragacanth; as well as carbohydrate or protein fillers, including but not limited to proteins such as gelatin and collagen. Disintegrants or solubilizers such as cross-linked polyvinylpyrrolidone, agar, alginic acid, or salts of alginic acid such as sodium alginate may be added as needed.

[0130] The sugar-coated tablet core is provided with suitable coatings such as concentrated sugar solutions that may also contain gum arabic, talc, polyvinylpyrrolidone, Carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablet or sugar-coated tablet coating for product identification or to characterize the amount of the active compound (i.e., dosage). The pharmaceutical preparations disclosed herein may be used orally, for example, using gelatin push-fit capsules, as well as gelatin soft seal capsules, and coatings such as glycerol or sorbitol. The push-fit capsules may contain the compound of formula I mixed with a filler or binder such as lactose or starch, a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the compound of formula I may be dissolved or suspended in a suitable liquid, such as fatty oil, liquid paraffin, or liquid polyethylene glycol, with or without a stabilizer.

[0131] Liquid preparations include solutions, suspensions, and emulsions, such as water and water / propylene glycol solutions. For parenteral injection, liquid preparations may be formulated in solution in an aqueous polyethylene glycol solution.

[0132] Aqueous solutions suitable for oral use can be prepared by dissolving the active ingredient in water and adding appropriate colorants, fragrances, stabilizers, and thickeners as needed. Aqueous suspensions suitable for oral use can be prepared by dispersing the micronized active ingredient in water using viscous materials such as natural or synthetic rubber, resins, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth rubber, and acacia rubber, as well as dispersants or wetting agents such as naturally occurring phosphatides (e.g., lecithin), condensation products of alkylene oxides and fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide and long-chain aliphatic alcohols (e.g., heptadecaethyleneoxycetanol), condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous suspension may further contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavorings, and one or more sweeteners such as sucrose, aspartame, or saccharin. The formulation can be adjusted for volumetric osmolality.

[0133] This also includes solid-state preparations intended to be converted into liquid preparations for oral administration immediately before use. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active ingredient, colorants, flavorings, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.

[0134] Oily suspensions can be formulated by suspending the compound of Formula I in a vegetable oil such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin, or in a mixture thereof. Oily suspensions may contain thickeners such as beeswax, solid paraffin, or cetyl alcohol. Sweeteners such as glycerol, sorbitol, or sucrose can be added to provide an oral preparation with a pleasant mouthfeel. These formulations can be preserved by the addition of antioxidants such as ascorbic acid. For an example of an injectable oily vehicle, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997, which, in whole, is incorporated herein by reference with respect to all teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein. The pharmaceutical compositions disclosed herein may also be in the form of oil-in-water emulsions. The oil phase may be the vegetable oil or mineral oil described above, or a mixture thereof. 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 such as sorbitan monooleate and anhydrous hexitol, and condensates of partial esters having ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may further contain sweeteners and flavorings, as in the case of syrup and elixir formulations. Such formulations may further contain lubricants, preservatives, or colorants.

[0135] Pharmaceutical formulations of the compounds of formula I disclosed herein can be provided as salts and can be formed with bases, i.e., cationic salts, such as alkali and alkaline earth metal salts, such as sodium, lithium, potassium, calcium, and magnesium, as well as ammonium salts, such as ammonium, trimethylammonium, diethylammonium, and tris(hydroxymethyl)methylammonium salts.

[0136] Pharmaceutical preparations are preferably in unit dosage forms. In such forms, the preparation is subdivided into unit doses containing an appropriate amount of the active ingredient. A unit dosage form may be a packaged preparation, and the packaging contains separate amounts of the preparation, such as packaged tablets, capsules, and powders in vials or ampoules. Furthermore, a unit dosage form may be a capsule, tablet, cachet, or lozenge itself, or an appropriate number of any of these in packaged form.

[0137] The amount of the active ingredient in a unit dose preparation may vary or be adjusted from 0.1 mg to 10,000 mg, more typically from 1.0 mg to 1,000 mg, and most typically from 10 mg to 500 mg, depending on the specific use and the potency of the active ingredient. The composition may further contain other suitable therapeutic agents if desired.

[0138] The administration regimen should also take into account pharmacokinetic parameters well known in the art, namely absorption rate, bioavailability, metabolism, and clearance (e.g., Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci. 84:1144-1146; Rohatagi (1995) Pharmazie See 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24:103-108; and the latest Remington's (above), each of which, with reference to all of its teachings, including but not limited to all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein, is incorporated herein by reference in their entirety. State-of-the-art technology enables clinicians to determine the dosage regimen for each individual patient, GR and / or MR modulator, and each disease or illness being treated.

[0139] A single or multiple dose of a compound formulation of formula I can be administered according to the required and tolerated dosage and frequency by the patient. The formulation should provide a sufficient amount of the active agent to effectively treat the disease condition. Accordingly, in one embodiment, a pharmaceutical formulation for oral administration of a compound of formula I is a daily dose of about 0.5 to about 30 mg / kg body weight / day, including all partial ranges and partial values ​​therein, including the endpoint. In an alternative embodiment, the dose is about 1 mg to about 20 mg / kg body weight / patient / day. Lower doses can be used, in particular, when the drug is administered into an anatomically occluded site such as the cerebrospinal fluid (CSF) space, into the bloodstream, into a body cavity, or into the lumen of an organ, in contrast to oral administration. Substantially higher doses can be used for topical administration. Practical methods for preparing formulations containing a compound of formula I for parenteral administration are known or evident to those skilled in the art and are described in detail in publications such as Remington's (above). See also Nieman, In “Receptor Mediated Antisteroid Action”, Agarwal, et al., eds., De Gruyter, New York (1987), which, in whole, is incorporated herein by reference with respect to all of its teachings, including, but not limited to, all methods, compounds, compositions, data, etc., for use in conjunction with any of the embodiments and disclosures herein.

[0140] In some embodiments, simultaneous administration includes administering one active agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours (or any sub-range of time or sub-value of time within 24 hours) of the second active agent. Simultaneous administration includes administering two active agents simultaneously, nearly simultaneously (e.g., within approximately 1, 5, 10, 15, 20, or 30 minutes of each other (or, e.g., any sub-range of time or sub-value of time between 0 and 30 minutes)), or sequentially in any order. In some embodiments, simultaneous administration can be achieved by simultaneous formulation, i.e., by preparing a single pharmaceutical composition containing both active agents. In some embodiments, the active agents may be formulated separately. In some embodiments, the active agents and / or adjuvants may be linked or conjugated with each other. At least one dose of the drugs may be administered, for example, simultaneously. At least one dose of the drugs may be administered, for example, within a few minutes or less than an hour of each other. At least one dose of the drug can be administered, for example, at different times, but on the same day or on different days.

[0141] A pharmaceutical composition comprising a compound of formula I disclosed herein may, after being formulated in one or more acceptable carriers, be placed in a suitable container and labeled for treatment of the indicated conditions. Such labeling for the administration of a compound of formula I may include, for example, instructions regarding the amount, frequency, and method of administration.

[0142] Drug administration The administration regimens of the compounds described herein will naturally vary depending on known factors such as the pharmacodynamic characteristics of the particular drug and its mode and route of administration; the recipient's species, age, sex, health status, medical condition, and weight; the nature and severity of symptoms; the type of concurrent treatment; the frequency of treatment; the route of administration, the patient's renal and hepatic function, and the desired effect. Clinicians can determine and prescribe the effective dose of the drug necessary to prevent, counteract, or halt the progression of a disease or disorder.

[0143] As a general guideline, the daily oral dose of each active ingredient, when used for the indicated effect, is in the range of about 0.001 to about 1000 mg / kg body weight, preferably about 0.01 to about 100 mg / kg body weight / day, and most preferably about 0.1 to about 20 mg / kg / day. In some embodiments, the compound of formula (I) may be administered in doses between about 10 mg / day and about 200 mg / day. In some embodiments, the compound of formula (I) may be administered at doses of approximately 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 110 mg / day, 120 mg / day, 130 mg / day, 140 mg / day, 150 mg / day, 160 mg / day, 170 mg / day, 180 mg / day, 190 mg / day, or 200 mg / day. The dose may be any value or a partial range within the ranges described in detail.

[0144] Depending on the patient's condition and the intended therapeutic effect, the frequency of administration of the therapeutic agent may vary, for example, from once to six times a day. That is, the frequency of administration may be QD, i.e., once a day; BID, i.e., twice a day; TID, i.e., three times a day; QID, i.e., four times a day; five or six times a day. In another embodiment, the frequency of administration may be BIW, i.e., twice a week; TIW, i.e., three times a week; or QIW, i.e., four times a week.

[0145] Depending on the patient's condition and the intended therapeutic effect, the treatment cycle may include periods during which the therapeutic agent is not administered. As used herein, “interval administration” refers to administration of the therapeutic agent and the subsequent gap of days or weeks. For example, a treatment cycle may be three weeks, including two weeks of therapeutic agent administration followed by one week without therapeutic agent administration. In some embodiments, a treatment cycle may be four weeks, including three weeks of administration followed by one week without therapeutic agent administration.

[0146] As used herein, the term “treatment cycle” refers to a predetermined period of time for administering a therapeutic agent. Typically, the patient is examined at the end of each treatment cycle to assess the effectiveness of the treatment.

[0147] In one embodiment, each treatment cycle has about 3 days or more. In another embodiment, each treatment cycle has about 3 days to about 60 days. In another embodiment, each treatment cycle has about 5 days to about 50 days. In another embodiment, each treatment cycle has about 7 days to about 28 days. In another embodiment, each treatment cycle has 28 days. In one embodiment, the treatment cycle has about 29 days. In another embodiment, the treatment cycle has about 30 days. In another embodiment, the treatment cycle has about 31 days. In another embodiment, the treatment cycle has a treatment cycle of about 1 month. In another embodiment, the treatment cycle is of any length from 3 weeks to 8 weeks. In another embodiment, the treatment cycle is of any length from 3 weeks to 6 weeks. In yet another embodiment, the treatment cycle is 3 weeks. In another embodiment, the treatment cycle is 1 month. In another embodiment, the treatment cycle is 4 weeks. In another embodiment, the treatment cycle is 5 weeks. In another embodiment, the treatment cycle is 6 weeks. In another embodiment, the treatment cycle is 7 weeks. In another embodiment, the treatment cycle is 8 weeks. The duration of the treatment cycle may include any value or partial range within the detailed range, including the endpoint.

[0148] As used herein, the terms “co-administration” or “co-administration” refer to the coordinated administration of (a) an additional therapeutic agent and (b) a compound of formula (I), or its salts, solvates, esters, and / or prodrugs. For example, co-administration may be simultaneous administration, sequential administration, overlapping administration, interval administration, consecutive administration, or a combination thereof.

[0149] In some embodiments, the administration regimen for the compound of formula (I) is once daily for a continuous 28-day cycle. In some embodiments, the once-daily administration regimen for the compound of formula (I) may be, but are not limited to, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, or 60 mg / day. The compound of formula (I) may be administered once daily in any dose from 20 mg to 60 mg. The dose may be any value or a partial range within the detailed range.

[0150] In some embodiments, the administration regimen for the compound of formula (I) is twice daily for a continuous 28-day cycle. In some embodiments, the twice-daily administration regimen for the compound of formula (I) may be, but are not limited to, 10 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, or 100 mg / day. The compound of formula (I) may be administered twice daily in any dose from 20 mg to 80 mg. In some embodiments, the compound of formula (I) may be administered in any dose from 10 mg / day to 100 mg / day. The dose may be any value or a partial range within the detailed range.

[0151] In some embodiments, the administration regimen of the compound of formula (I) may be 20 mg to 60 mg / day once daily for two weeks, followed by a one-week break over six weeks (e.g., two weeks on, one week off). In some embodiments, the administration regimen of the compound of formula (I) may be 10 mg to 100 mg twice daily for two weeks, followed by a one-week break over six weeks (e.g., two weeks on, one week off).

[0152] In some embodiments, the administration regimen for the compound of formula (I) may be 20 mg to 60 mg / day once daily for 3 weeks, followed by a 1-week break over 8 weeks (e.g., 3 weeks on, 1 week off). In some embodiments, the administration regimen for the compound of formula (I) may be 10 mg to 100 mg twice daily three times daily for 3 weeks, followed by a 1-week break over 8 weeks (e.g., 8 weeks on, 1 week off).

[0153] In some embodiments, the administration regimen of the compound of formula (I) may be twice daily on days 1 and 2, weekly for 8 weeks. In some embodiments, the dosage of the compound of formula (I) may be, but is not limited to, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg.

[0154] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a three-week cycle consisting of two weeks of compound administration followed by one week of compound non-administration.

[0155] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered once daily in a four-week cycle including three weeks of compound administration followed by one week of compound non-administration.

[0156] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of 6 weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of 8 weeks.

[0157] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered three times a week. In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered on the first, third, and fifth days of the week.

[0158] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered four times a week.

[0159] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a three-week cycle consisting of two weeks of compound administration followed by one week of compound non-administration.

[0160] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is administered in a 4-week cycle including 3 weeks of compound administration followed by 1 week of compound non-administration.

[0161] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered twice daily, two days a week. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered over a period of eight weeks. In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is administered on the first and second days of each week.

[0162] When the compound of formula I is administered multiple times a week, the dosage may be administered on any day or combination of days within the week. For example, a three-times-a-week administration may include administrations on days 1, 3, and 5; days 1, 2, and 3; days 1, 3, and 5, etc. A two-days-a-week administration may include administrations on days 1 and 2; days 1 and 3; days 1 and 4; days 1 and 5; days 1 and 6; days 1 and 7, etc.

[0163] Kits and products Some embodiments relate to kits and products comprising a compound of formula I and / or at least one EGFR inhibitor. For example, a kit or product may include a package or container containing the compound of formula I. Such a kit and product may further include a product attachment or label with approved drug administration and instruction information, including how to use the compound of formula I in combination with an EGFR inhibitor provided separately. The kit can be used in methods of treating cancer as described herein.

[0164] In some embodiments, the kit or product may include both a compound of formula I and at least one EGFR inhibitor. In some embodiments, the EGFR inhibitor is, for example, osimertinib. Such a kit may include one or more containers or packages, one or more containers or packages containing one or both combination drugs together in a single container and / or package, or in separate packages / containers. In some examples, the two drugs are packaged separately but contained in a single package, container or box. Such kits and products may further include approved drug administration and instruction information, including how to use the compound of formula I in combination with the provided EGFR inhibitor, in a product attachment or label. The kit may be used in methods of treating cancer as described herein. [Examples]

[0165] Basic Procedure All starting materials and solvents were obtained from commercial sources or prepared according to literature references.

[0166] Example 1 Combination Cell Growth Assay: Cells (2000 cells per well) were seeded onto a 96-well plate in 100 μl of cell culture medium. Cells were treated with the compound of Formula I and osimertinib at concentrations ranging from 0 to 10 μM using the Tecan D300e Digital Dispenser combination matrix protocol. On day 5, 50 μl of CellTiter-Glo (CTG) reagent (Promega) was added, and the plate was incubated for 10 minutes with gentle shaking. After 10 minutes of incubation, the luminescence signal was determined according to the provider's instructions (Promega), and combination data was generated using the standard HSA model with Combinenefit software. Combination synergies are represented by positive numbers in the results table. Negative numbers represent combination antagonists.

[0167] Figures 1A and 1B show data demonstrating that the combined compound of formula I and the EGFR inhibitor osimertinib exhibit synergistic effects in vitro in the EGFR mutant cell line HCC827, lung adenocarcinoma with acquired mutations in the EGFR tyrosine kinase domain (E746-A750 deletion), and the NCI-H820 lung adenocarcinoma cell line with acquired mutations in the EGFR tyrosine kinase domain (T790M). Figure 1A shows numerical data for the compound of formula I and the EGFR inhibitor osimertinib in the cell line HCC827. Figure 1B shows numerical data for the compound of formula I and the EGFR inhibitor osimertinib in the cell line NCI-H820. The combination of the compound of formula I and EGFRi (osimertinib) shows a strong synergistic survival effect.

[0168] Example 2 The results from these experiments are summarized in Figures 3-6 and explained further below.

[0169] [Table 1]

[0170] Figures 2A and 2B show treatment of CAL-27 cells with either the compound of formula I alone, cetuximab alone, or a combination of the compound of formula I and cetuximab. Figure 2A shows a plot of activity percentage versus inhibitor concentration (log M) in CAL-27 cells treated with the compound of formula I alone (black circle, line 1) and in combination with cetuximab at 0.5 μg / ml (black square, line 2), 1.0 μg / ml (black circle, line 3), or 2.5 μg / ml (black square, line 4).

[0171] Figure 2B shows a bar graph of CTG activity percentages, indicating that cetuximab treatment alone reduced cell viability by approximately 40%.

[0172] [Table 2]

[0173] Figures 3A and 3B show SCC-9 cell treatment with either the compound of formula I alone, cetuximab alone, or a combination of the compound of formula I and cetuximab. Figure 3A shows a plot of activity percentage versus inhibitor concentration (log M) in SCC-9 cells treated with the compound of formula I alone and in combination with cetuximab at 0.5 μg / ml (black square, line 2), 1.0 μg / ml (black circle, line 3), or 2.5 μg / ml (black square, line 4). Figure 3B shows a bar graph of CTG activity percentage, indicating that cetuximab treatment alone reduced cell viability by approximately 15%.

[0174] [Table 3]

[0175] Figures 4A and 4B show treatment of SCC-15 cells with either the compound of formula I alone, cetuximab alone, or a combination of the compound of formula I and cetuximab. Figure 4A shows a plot of activity percentage versus inhibitor concentration (log M) in SCC-15 cells treated with the compound of formula I alone and in combination with cetuximab at 1.0 μg / ml (black square, line 2), 2.5 μg / ml (black circle, line 3), or 5.0 μg / ml (black square, line 4). Figure 4B shows a bar graph of CTG activity percentage, indicating that cetuximab treatment alone reduced cell viability by less than 10%.

[0176] [Table 4]

[0177] Figures 4A and 5B show treatment of SCC-25 cells with either the compound of formula I alone, cetuximab alone, or a combination of the compound of formula I and cetuximab. Figure 5A shows a plot of activity percentage versus inhibitor concentration (log M) in SCC-25 cells treated with the compound of formula I alone and in combination with cetuximab at 2.5 μg / ml (black square, line 2), 5.0 μg / ml (black circle, line 3), or 10.0 μg / ml (black square, line 4). Figure 5B shows a bar graph of CTG activity percentage, indicating that cetuximab treatment alone did not reduce cell viability.

[0178] [Table 5]

[0179] Figures 6A and 6B show the inhibition of ERK1 / 2 phosphorylation in HPV-negative CAL27 cells by either the compound of formula I alone, cetuximab alone, or a combination of the compound of formula I and cetuximab. Figure 6A shows an immunoblot of approximately 50% inhibition of ERK1 / 2 phosphorylation activity with the compound of formula I versus DMSO. Figure 6B shows the quantified phosphorylated ERK1 / 2 bands normalized by total ERK. Quantitative results showed that treatment with the compound of formula I or cetuximab alone reduced ERK1 / 2 phosphorylation by approximately 50% compared to DMSO-treated control cells, while the combination of the compound of formula I and cetuximab showed approximately 80% inhibition of ERK1 / 2 phosphorylation in the HPV-negative head and neck squamous cell carcinoma cell line CAL27.

[0180] Example 3: The combination of the compound of formula I and cetuximab showed a combined effect on MAPK signaling and cell viability in HPV-negative head and neck squamous cell carcinoma cells. The purpose of this study is to determine the combined effect of the compound of formula I on oncogenic MAPK signaling via SHP2 inhibition and on EGFR signaling via EGFR inhibition by cetuximab.

[0181] The cell lines used in this study were obtained from ATCC (CAL27#CRL-2095, SCC-15#CRL-1623, SCC-25#CRL-1628, SCC-4#CRL-1624, and SCC-9#CRL-1629). Cell line CAL27 was cultured in DMEM (Gibco#). SCC-4, SCC-15, SCC-25, and SCC-9 cell lines were cultured in a 1:1 mixture of DMEM and Ham's F12 medium containing 1.2 g / L sodium bicarbonate, 2.5 mM glutamine, 15 mM HEPES, and 400 ng / ml hydrocortisone, supplemented with 90% 0.5 mM sodium pyruvate; in fetal bovine serum, 10% (Hyclone#SH-30071.03) and penicillin / streptomycin (Thermo Fisher#15070-063). Cells were maintained at 37°C / 5% CO2. Cells were washed in ice-cold PBS (Thermo Fisher Scientific #10010-023) and lysed in RIPA lysis and extraction buffer (Thermo Fisher Scientific #89900) supplemented with Halt phosphatase and protease inhibitors (Thermo Fisher Scientific #78445). The cell membrane was blocked against nonspecific proteins using bovine serum albumin BSA (Cell Signaling Tech #9998). The antibodies used for immunoblotting were as follows: phospho-p44 / 42 ERK1 / 2 (Cell Signaling Tech #4370), ERK2 antibody (Santa Cruz #sc-1647), anti-mouse IgG HRP-binding antibody (Cell Signaling Tech #7076), and anti-rabbit IgG HRP-binding antibody (Cell Signaling Tech #7074).

[0182] Cell proliferation assay: Cells (2000 cells per well) were seeded onto a 96-well plate in 100 μl of cell culture medium. Cells were treated with concentrations of compound I of formula I ranging from 0 to 10 pM and fixed cetuximab concentrations of 0.5, 1, 2.5, 5, and 10 μg / ml using the Tecan D300e Digital Dispenser combined matrix protocol. On day 5, 50 μl of CellTiter-Glo (CTG) reagent (Promega) was added, and the plate was incubated for 10 minutes with gentle shaking. After 10 minutes of incubation, the luminescence signal was determined according to the provider's instructions (Promega), and combined data was generated using Combinefit software.

[0183] Western blotting: Cells were lysed on ice for 10 minutes using Thermo Fisher RIPA lysis buffer containing protease and phosphatase inhibitors. Cells were centrifuged at 4°C for 10 minutes using a microcentrifuge. The supernatant was transferred to a pre-cooled microcentrifuge tube, and the protein concentration of the lysate was measured using the BCA method. Equal volumes of protein-rich cell lysate supernatant were used for immunoblotting.

[0184] The objective of this study was to determine the combined effects of the compound of formula I on oncogenic MAPK signaling via SHP2 inhibition and EGFR signaling via EGFR inhibition by cetuximab. Three HPV-negative head and neck squamous cell carcinoma cell lines, CAL27, SCC-4, SCC-9, SCC-15, and SCC-25, were divided into 96-well plates. After overnight incubation, the compound of formula I and cetuximab were added to the cells using the Tecan D300e Digital Dispenser combined matrix protocol. After 5 days of incubation, the CellTiter-Glo assay was performed to test the combined effect by increasing the sensitivity of the compound of formula I to activity upon addition of cetuximab. The combined data showed that cetuximab increased the sensitivity of the compound of formula I to HPV-negative head and neck squamous cell carcinoma cell lines (Figures 2-5).

[0185] CAL27 cells were treated with the compound of formula I alone and also in combination with cetuximab. The compound of formula I alone inhibited cell proliferation in CAL27 cells with an IC50 of 112 nM. Sensitivity to the compound of formula I increased when combined with cetuximab. Concurrent treatment with cetuximab sensitized the compound of formula I in a dose-dependent manner, and the IC50 of the compound of formula I decreased from 112 nM to 57 nM upon concurrent treatment with 2.5 μg / ml of cetuximab (Figure 2A and Table 2). Cetuximab monotherapy further reduced cell viability by approximately 40% (Figure 2B), suggesting that cetuximab monotherapy was effective in this cell line. The combination of the compound of formula I and cetuximab showed a more pronounced inhibition of cell viability.

[0186] SCC-9 cells were treated with the compound of formula I alone and in combination with cetuximab. The compound of formula I inhibited cell proliferation of SCC-9 cells with an IC50 of 161 nM. Sensitivity to the compound of formula I increased when combined with cetuximab. Concurrent treatment with cetuximab sensitized the compound of formula I in a dose-dependent manner, and the IC50 of the compound of formula I decreased from 161 nM to 36 nM with concurrent treatment with 2.5 μg / ml of cetuximab (Figure 3A and Table 3). However, no significant decrease in cell viability was observed with cetuximab monotherapy at up to 2.5 μg / ml (Figure 3B). The combination of the compound of formula I and cetuximab showed a more significant inhibition of cell viability.

[0187] SCC-15 cells were treated with the compound of formula I alone and in combination with cetuximab. The compound of formula I inhibited cell proliferation of SCC-15 cells with an IC50 of 1406 nM. Sensitivity to the compound of formula I increased when combined with cetuximab. Concurrent treatment with cetuximab sensitized the compound of formula I in a dose-dependent manner, and the IC50 of the compound of formula I decreased from 1406 nM to 55 nM with concurrent treatment with 5 μg / ml cetuximab (Figure 4A and Table 4). However, no significant decrease in cell viability was observed with cetuximab monotherapy at up to 5 μg / ml (Figure 4B). The combination of the compound of formula I and cetuximab showed a more significant inhibition of cell viability.

[0188] SCC-25 cells were treated with the compound of formula I alone and in combination with cetuximab. The compound of formula I inhibited cell proliferation in SCC-15 cells with an IC50 of 8046 nM. Sensitivity to the compound of formula I increased when combined with cetuximab. Concurrent treatment with cetuximab sensitized the compound of formula I in a dose-dependent manner, and the IC50 of the compound of formula I decreased from 1406 nM to 418 nM with concurrent treatment with 10 μg / ml cetuximab (Figure 5A and Table 5). However, no significant decrease in cell viability was observed with cetuximab monotherapy at up to 5 μg / ml (Figure 5B). The combination of the compound of formula I and cetuximab showed a more significant inhibition of cell viability.

[0189] The combination of formula I and cetuximab showed robust inhibition of ERK1 / 2 phosphorylation in HPV-negative CAL27 cells. CAL27 cell lines were divided into 6-well plates and treated separately with formula I alone, cetuximab alone, and in combination, as shown (Figures 6A-6B). Four hours after treatment, cells were lysed and immunoblotted for phosphorylated ERK1 / 2. Immunoblotting results showed that formula I inhibited ERK1 / 2 phosphorylation by approximately 50% compared to DMSO-treated control cells. Cetuximab treatment showed approximately 50% inhibition of ERK1 / 2 phosphorylation, and the inhibition was more pronounced (approximately 80%) when combined with formula I (Figure 6A). Phosphorylated ERK1 / 2 bands were quantified using Bio-Rad Image Lab software and normalized by total ERK. Quantitative results showed that treatment with compound I alone or cetuximab alone reduced ERK1 / 2 phosphorylation by approximately 50% compared to DMSO-treated control cells, while the combination of compound I and cetuximab showed approximately 80% inhibition of ERK1 / 2 phosphorylation in the HPV-negative head and neck squamous cell carcinoma cell line CAL27 (Figure 6B).

[0190] Example 4: Open-label Phase 1b / 2 study of the compound of the Disclosure in combination with other anticancer therapies in patients with advanced or metastatic solid tumors. Test design The compounds of this disclosure in the form of a pharmaceutical composition (e.g., the compound of Formula I) will be administered in combination with other cancer treatments to subjects with solid tumors exhibiting specific molecular changes in an open-label, multicenter clinical trial. After a screening period, eligible subjects will be enrolled and treated with the pharmaceutical composition containing the compound of Formula I and other anticancer treatments until disease progression, unacceptable toxicity, or other criteria for discontinuing treatment are met.

[0191] This study aims to evaluate the safety and tolerability of the escalating doses of the compounds disclosed herein (e.g., the compound of formula I) when administered in combination with other cancer therapies, to determine the maximum tolerated dose (MTD) and / or recommended dose (RD) of the compound of formula I when administered in combination with other cancer therapies, to characterize the pharmacokinetic (PK) profile of the compound when administered in combination with other cancer therapies, and to evaluate its antitumor activity when administered in combination with other cancer therapies.

[0192] Evaluation items The primary endpoints to be evaluated are: (1) Dose-limiting toxicity (DLT) (based on observed toxicity), (2) Maximum tolerated dose (MTD) (based on observed toxicity), (3) Recommended dose (RD) (based on observed toxicity), (4) Adverse events (AEs) (incidence and severity of treatment-related and serious AEs) (time frame: evaluated from the time of first administration to 24 months), (5) Plasma concentration (Cmax) (time frame: up to day 29 from the study date), (6) Time to reach Cmax (Tmax) (time frame: up to day 29 from the study date), (7) Area under the curve (area under the plasma concentration-time curve of the compound disclosed herein), and (8) Half-life (time frame: from day 1 to day 29 of the study).

[0193] Secondary endpoints to be evaluated: (9) Objective response rate (ORR) (based on X-ray image evaluation according to RECIST version 1.1) (Time frame: evaluated from the time of first administration to 24 months) (10) Duration of response (DOR) (based on X-ray image evaluation according to RECIST version 1.1) (11) Time to response (TTR) (based on X-ray image evaluation according to RECIST version 1.1) (Time frame: evaluated from the time of first administration to 24 months).

[0194] Other pre-specified evaluation items: (12) Pharmacodynamic evaluation (evaluation of phosphorylated ERK (pERK) inhibition in PBMCs or tumor tissue by IHC or immunofluorescence (time frame: evaluated from the first dose up to 24 months)).

[0195] Patient eligibility criteria for the study: (1) Age ≥ 18 years; (2) Willing and able to give written informed consent; (3) Histologically or cytologically confirmed advanced or metastatic solid tumor; (4) No available standard systemic therapy for the patient's tumor histology and / or molecular biomarker profile, or standard therapy is tolerable, ineffective, or inaccessible; (5) Able to swallow oral medication; (6) Eastern Coordinated Oncology Group Performance Status (ECOG PS) of 0 or 1; (7) Having adequate cardiovascular, hematological, hepatic, and renal function; and (7) Willing to follow visits, assessments, and procedures required by all protocols.

[0196] Exclusion criteria: (1) prior treatment with an SHP2 inhibitor; (2) recorded PTPN11 mutation; (3) receiving another study therapy or participating in a study of the investigational drug within 4 weeks of the first dose; (4) receiving pre-palliative radiation on day 1 of cycle 1, day 7 or less; (5) primary central nervous system disease or known active CNS metastasis and / or carcinomatous meningitis; (6) having pre-operative or gastrointestinal dysfunction that may affect drug absorption; (7) clinically significant interstitial lung disease or pneumonia; (8) history of thromboembolism or cerebrovascular event within 12 weeks prior to the first dose; (9) history or current evidence of retinal vein occlusion (RVO or current risk factor for RVO); (10) having any underlying medical condition, mental illness or social condition that, in the opinion of the principal investigator, could impair study administration according to the protocol or assessment of Aes; and (11) being pregnant or lactating, or expecting to become pregnant or father during the study period.

[0197] HPV-negative, head and neck squamous cell carcinoma: For once-daily consecutive doses (QD), the dose levels are 40 mg, 20-60 mg QD, 40 mg QD, or 60 mg QD for the compound of formula I. For twice-daily consecutive doses (BID), the dose levels are 10-100 mg BID or 20-80 mg BID. The planned dosing schedule for QD or BID is 2 weeks on / 1 week off. Cetuximab is administered every other week.

[0198] wtKRAS / wtNRAS / wtBRAF, colorectal cancer: For once-daily consecutive administration (QD), the dose levels are 40 mg, 20-60 mg QD, 40 mg QD, or 60 mg QD for the compound of formula I. For twice-daily consecutive administration (BID), the dose levels are 10-100 mg BID or 20-80 mg BID. The planned administration schedule for QD or BID is 2 weeks on / 1 week off. Cetuximab is administered every other week.

[0199] Example 5 - Combination therapy of compound I and osimertinib in the HCC827 cell line and the erlotinib-resistant cell line HCC827 / ER1 material and method The human NSCLC cell line, HCC827, was purchased from ATCC. EGFR exon 19 deletion mutant and MET-amplified HCC827 / ER1 cell lines (Crown Bioscience UK) were induced from HCC827 (ATCC) at Crown Bioscience by culturing cells in the presence of escalating concentrations of erlotinib. HCC827 / ER1 cells were cultured at 37°C in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and 42 pM erlotinib under 5% CO2 air. The medium was conventionally refreshed, and tumor cells were subcultured every 3–5 days at 80% confluence with trypsin-EDTA (Moore et al. 2016b).

[0200] Cell proliferation assay: Cells were collected during the logarithmic growth period and counted using Count-star. Cells were placed on a 96-well plate in a 4 × 10⁶ arrangement. 3Cells were seeded at a final cell density of cells / mL in a volume of 100 μL / well. After incubation overnight, plated cells were equilibrated at room temperature (RT) for approximately 30 minutes. CellTiter-Glo reagent (50 μL) was added to each well and mixed on an orbital shaker for 5 minutes to induce cell lysis. The plate was incubated at RT for 20 minutes to stabilize the luminescence signal. The T0 luminescence signal was measured using an EnVision Multi Label Reader. For concomitant treatment, drugs were prepared and simultaneously distributed in 1000 × drug solutions of each test item in each well. The plate was incubated in a humidified incubator at 37°C and 5% CO2 for 120 hours, and then measured by CTG assay. The plate was equilibrated at room temperature for approximately 30 minutes. CellTiter-Glo (50 μL) was added to each well, and the plate was incubated on an orbital shaker for 5 minutes to induce cell lysis. The plates were incubated at room temperature for 20 minutes to stabilize the luminescence signal (T5), and then measured using an EnVision Multi Label Reader.

[0201] Synergistic effect calculation The synergy score was calculated using the R package "SynergyFinder" based on either the Loewe additive model or the Bliss independence model. Details of the calculation method are described below.

[0202] In a drug combination experiment where drug A is administered at dose x1 and drug B is administered at dose x2, the effect of this combination is measured as EABobs. In addition, the effect of drug A at dose x1 is EA, and the effect of drug B at dose x2 is EB.

[0203] The expected drug effect (EAB) is calculated based on either the Bliss model or the Loewe model:

[0204] Bliss model:EAB=EA+EB-EA*EB

[0205] Loewe Model: A four-parameter log-logistic model was used to fit the dose-response curves of each drug, and then the EAB was calculated based on the parameters of the log-logistic model using a nonlinear equation.

[0206] Figure 7A shows a table of Bliss synergy scores for combinations of the compound of formula I with osimertinib in the (erlotinib-resistant) HCC827 / ER1 cell line.

[0207] Figure 7B shows a table of Bliss synergy scores for combinations of the compound of formula I and osimertinib in the (erlotinib-resistant) HCC827ER1 parent cell line.

[0208] The difference in proportions between EABobs and EABs represents the synergistic score of that combination.

[0209] Example 6 - Combination therapy of compound I and osimertinib in EGFR L858R / T790M mutant NSCLC PDX model LUN2355-215 material Vehicle / control and 100 mM acetic acid in deionized water were prepared by adjusting the pH to 4.8-5.0, and stored under ambient conditions for 18 days during administration to mice.

[0210] The test substance compound of Formula 1 was prepared weekly in a vehicle of 100 mM acetate buffer and stored under ambient conditions. The combination drug osimertinib was prepared weekly in a vehicle of 0.5% HPMC and 0.1% Tween 80 and stored under ambient conditions.

[0211] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were housed in a specially sourced pathogen-free (SPF) environment and allowed to acclimate to the new environment for at least three days before the start of any experiment. The mice were 6-8 weeks old at the time of transplantation.

[0212] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Animal Care Committee (IACUC). The animal facilities and programs operated under the standards of the Guidelines for the Management and Use of Laboratory Animals (NRC, 2011) and were accredited by the International Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0213] Preparation of PDX models The LUN2355-215 model was established for preclinical efficacy studies at GenenDesign (Shanghai, China). This PDX model was derived from a 49-year-old male Chinese NSCLC patient. EGFR mutations in the PDX model were confirmed by whole exom sequencing and PCR sequencing. Tumor fragments taken from the PDX model were subcutaneously transplanted into the right flank of female Balb / c nude mice. Mice were anesthetized with isoflurane and anesthesia was maintained throughout the transplantation surgery. The skin of the mice was cleaned on the right flank with appropriate surgical scrub and alcohol. A small skin incision was made using the sharp end of a trocar, and a 1.5 cm subcutaneous pocket along the right chest wall was formed by blunt incision with a 10-12 g trocar needle stylet. Tumor fragments (15-30 mm) 3 The trocar needle was inserted and advanced into the subcutaneous pocket in the right flank. The trocar incision was closed with sutures or wound clips, which were removed one week after closure.

[0214] Tumor size is 150-250 mm in volume 3 When the target was reached, mice with tumors were randomly divided into test groups, with 8 mice in each group. The randomization day is denoted as treatment day 0.

[0215] treatment Treatment was initiated on the same day as randomization. The treatment start day was denoted as treatment day 0. Mice were administered orally with a vehicle control solution, the compound of formula I alone at 15 mg / kg QD, osimertinib alone at 15 mg / kg QD, and a combination of the compound of formula I and osimertinib at 15 mg / kg QD. The dose was 5 mL / kg for each compound. In the combination group, osimertinib was administered 1 hour after the administration of the compound of formula I QD. This study was concluded on treatment day 18 as defined in the study protocol.

[0216] result Figure 8 shows graphs of tumor volume over treatment periods with regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR L858R / T790M mutant NSCLC PDX model LUN2355-215. No significant changes in body weight were observed in either the control or treatment groups.

[0217] conclusion As shown in Figure 8, the combination of compound I and osimertinib demonstrated superior tumor growth inhibition in the EGFR L858R / T790M mutant NSCLC PDX model LUN2355-215 compared to treatment with compound I alone or osimertinib alone.

[0218] Example 7 - Combination therapy of compound I and osimertinib in osimertinib-resistant MET-amplified NSCLC CDX model NCI-H820 material Vehicle / control and 100 mM acetic acid in deionized water were prepared by adjusting the pH to 4.8-5.0, and stored under ambient conditions for 28 days during administration to mice.

[0219] Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetate buffer and stored under ambient conditions. The combination drug osimertinib was prepared weekly in a vehicle of 0.5% HPMC and 0.1% Tween 80 and stored under ambient conditions.

[0220] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were 6-8 weeks old at the time of transplantation. The mice were housed in a specially sourced pathogen-free (SPF) environment and allowed to acclimate to the new environment for at least 3 days before the start of any experiment.

[0221] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with guidelines approved by the WuXiAppTec Animal Care Committee (IACUC). During the study, animal care and use were carried out in accordance with the regulations of the International Association for Accreditation and Evaluation of Laboratory Animal Care (AAALAC). In addition, all parts of this study conducted at WuXiAppTec adhered to the research protocol made and approved by the principal investigator and applicable standard operating procedures (SOPs).

[0222] Preparation of xenograft models NCI-H820 is a human lung adenocarcinoma cell line possessing the EGFR exon 19 deletion mutation, the EGFR T790M mutation, and MET amplification. The NCI-H820 cell line was purchased from the American Cell Culture and Cell Preservation Center (ATCC® HTB-181®). NCI-H820 cells were cultured at 37°C in RPML1640 medium containing 10% fetal bovine serum (FBS) and 1% antibiotic-antifungal agent (AA) under 5% CO2 atmosphere. The medium was refreshed every 2-3 days, and tumor cells were routinely subcultured at 80-90% confluence with trypsin-EDTA. Cells growing in the exponential growth phase were collected and counted for inoculation.

[0223] NCI-H820 tumor cells were subcutaneously transplanted into mice. In short, 10x10 cells mixed with 50% Matrigel were transplanted. 6A 200 μL cell suspension containing tumor cells was subcutaneously transplanted into the right flank of mice using a syringe. The animals' health and tumor growth were monitored daily. Tumor volume was measured twice weekly using a caliper if the tumor was palpable and measurable. The average tumor volume was 171 mm². 3 (96~251mm 3 When the tumor reached a certain level, mice with tumors were randomized into different groups of 8 mice each. The randomization day is denoted as treatment day 0.

[0224] treatment Treatment was initiated on the day after randomization. The treatment initiation day is denoted as treatment day 1. Mice were administered orally with vehicle control solution, compound I alone at 30 mg / kg QD, and osimertinib alone at 15 mg / kg QD. One additional group received combination treatment with compound I and osimertinib, administration of compound I at 15 mg / kg QD, and osimertinib at 15 mg / kg QD. The dose was 5 mL / kg for each compound. In the combination group, osimertinib was administered 1 hour after administration of compound I QD. This study was concluded on treatment day 28 as defined in the study protocol.

[0225] result Figure 9 shows graphs of tumor volume over treatment periods for regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR delE746_E749 / T790M mutant and the MET-amplified NSCLC CDX model NCI-H820. No significant changes in body weight were observed in the control or treatment groups.

[0226] conclusion As shown in Figure 9, the combination of the compound of formula I with osimertinib demonstrated superior tumor growth inhibition in EGFR delE746_E749 / T790 and MET-amplified NSCLC CDX model NCI-H820 compared to treatment with the compound of formula I alone or osimertinib alone.

[0227] Example 8 - Combination therapy with compound of formula I and osimertinib in the EGFR L858R / T790M mutant and ERBB2 overexpressing NSCLC PDX model LUN2005-143-9 material Acetic acid in 100 mM water was prepared for the vehicle / control, with the pH adjusted to 4.8-5.0, and stored under ambient conditions for 28 days during administration to mice.

[0228] The test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetate buffer and stored under ambient conditions. The combination drug osimertinib was prepared weekly in a vehicle of 0.5% HPMC and 0.1% Tween 80 and stored under ambient conditions.

[0229] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were housed in a specially sourced pathogen-free (SPF) environment and allowed to acclimate to the new environment for at least three days before the start of any experiment. The mice were 6-8 weeks old at the time of transplantation.

[0230] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Animal Care Committee (IACUC). The animal facilities and programs operated under the standards of the Guidelines for the Management and Use of Laboratory Animals (National Research Council, 2011) and were accredited by the International Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0231] Preparation of PDX models The LUN2005-143-9 model was established for preclinical efficacy studies at GenenDesign (Shanghai, China). The parent PDX model was derived from a 60-year-old male Chinese NSCLC patient with a tumor bearing the EGFR L858R mutation. LUN2005-143-9, an osimertinib-resistant tumor with high ERBB2 expression, was derived from approximately 60 mice with parent tumors approximately 8 months after treatment with osimertinib at 15 mg / kg QD. Tumor fragments taken from the PDX model were subcutaneously transplanted into the right flank of female Balb / c nude mice. To do this, the mice were anesthetized with isoflurane and anesthesia was maintained throughout the transplantation surgery. Using sterile surgical techniques, the skin of the mice was cleaned on the right flank with appropriate surgical scrub and alcohol. A small skin incision was made using the sharp end of a trocar, and a 1.5 cm subcutaneous pocket along the right chest wall was formed by blunt incision with a 10-12 g trocar needle stylet. Tumor fragments (15-30 mm) 3 The tumor was inserted into a trocar needle and advanced into a subcutaneous pocket in the right flank. The trocar incision was closed with sutures or wound clips and removed one week after closure. The tumor size was 150-250 mm in volume. 3 When the target was reached, mice with tumors were randomly divided into test groups, with 8 mice in each group. The randomization day is denoted as treatment day 0.

[0232] treatment Treatment was initiated on the same day as randomization. The treatment start day was denoted as treatment day 0. Mice were administered orally by vehicle control solution, osimertinib alone at 15 mg / kg QD, and the compound of formula I alone at 15 mg / kg QD. One additional group accepted combination treatment with the compound of formula I and osimertinib, receiving either the compound of formula I at 15 mg / kg QD or osimertinib at 15 mg / kg QD. The dose was 5 mL / kg for each compound. In the combination group, osimertinib was administered 1 hour after the QD dose of the compound of formula I. This study was concluded on treatment day 28 as defined in the study protocol.

[0233] result Figure 10 shows graphs of tumor volume over the treatment period with regimens of the compound of formula I alone, osimertinib alone, and a combination of the compound of formula I and osimertinib in an EGFR L858R variant and ERBB2-overexpressing NSCLC PDX model LUN2005-143-9. No significant body weight changes were observed in the control and treatment groups.

[0234] Conclusion As shown in Figure 10, the combination of the compound of formula I and osimertinib showed superior tumor growth inhibition in the EGFR L858R variant and ERBB2-overexpressing NSCLC PDX model LUN2005-143-9 compared to treatment with the compound of formula I alone or osimertinib alone.

[0235] Example 9 - Combination Therapy of the Compound of Formula I and Osimertinib in an EGFR L858R Variant NSCLC PDX Model LUN2005-234 Materials The vehicle / control, 100 mM acetic acid in deionized water, was prepared by adjusting the pH to 4.8 - 5.0 and stored under ambient conditions during the 24-day dosing period in mice.

[0236] The test article of the compound of formula I was prepared in a vehicle of 100 mM acetic acid buffer weekly and stored under ambient conditions. The combination drug osimertinib was prepared weekly in a vehicle of 0.5% HPMC and 0.1% Tween 80 and stored under ambient conditions.

[0237] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were housed in a specific pathogen-free (SPF) environmental breeding facility and acclimated to their new environment for at least 3 days before the start of any experiment. The mice were 6 - 8 weeks old at the time of transplantation.

[0238] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Animal Care Committee (IACUC). The animal facilities and programs operated under the standards of the Guidelines for the Management and Use of Laboratory Animals (National Research Council, 2011) and were accredited by the International Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0239] Preparation of PDX models The LUN2005-234 model was established for preclinical efficacy studies at GenenDesign (Shanghai, China). The parent PDX model was derived from a 60-year-old male Chinese NSCLC patient with a tumor bearing the EGFR L858R mutation. Osimertinib-resistant tumor, LUN2005-234, was derived from approximately 60 mice with parent tumors approximately 7 months after treatment with osimertinib at 15 mg / kg QD. Tumor fragments harvested from the PDX model were subcutaneously transplanted into the right flank of female Balb / c nude mice. To do this, the mice were anesthetized with isoflurane and anesthesia was maintained throughout the transplantation surgery. Using sterile surgical techniques, the skin of the mice was cleaned on the right flank with appropriate surgical scrub and alcohol. A small skin incision was made using the sharp end of a trocar, and a 1.5 cm subcutaneous pocket along the right chest wall was formed by blunt incision with a 10-12 g trocar needle stylet. Tumor fragment (15-30 mm) 3 The trocar was inserted into a trocar needle and advanced into a subcutaneous pocket in the right flank. The trocar incision was closed with sutures or wound clips and removed one week after closure. The average tumor volume was 205 mm². 3 When it reaches (tumor size is 150-250 mm) 3 Mice with tumors (within the specified range) were randomly divided into test groups of 8 mice each. The randomization day is denoted as treatment day 0.

[0240] treatment Treatment was initiated on the same day as randomization. The treatment start day was denoted as treatment day 0. Mice were administered orally by vehicle control solution, osimertinib alone at 15 mg / kg QD, and the compound of formula I alone at 15 mg / kg QD. One additional group accepted combination treatment with the compound of formula I and osimertinib, receiving the compound of formula I at 15 mg / kg QD and osimertinib at 15 mg / kg QD. The dose was 5 mL / kg for each compound. In the combination group, osimertinib was administered 1 hour after the QD dose of the compound of formula I. This study was concluded on treatment day 24 as defined in the study protocol.

[0241] result Figure 11 shows graphs of tumor volume over treatment periods for regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFR L858R mutant NSCLC PDX model LUN2005-234. No significant changes in body weight were observed in either the control or treatment groups.

[0242] conclusion As shown in Figure 11, the combination of the compound of formula I with osimertinib demonstrated superior tumor growth inhibition in the EGFR L858R mutant NSCLC PDX model LUN2005-234 compared to treatment with the compound of formula I alone or osimertinib alone.

[0243] Example 10 - Combination therapy of compound I and osimertinib in EGFR L858R / T790M mutant NSCLC PDX model LUN2355-128-33 material Acetic acid in 100 mM water was prepared for the vehicle / control, with the pH adjusted to 4.8-5.0, and stored under ambient conditions for 20 days during administration to mice.

[0244] Test samples of the compound of formula I were prepared weekly in a vehicle of 100 mM acetate buffer and stored under ambient conditions. The combination drug osimertinib was prepared weekly in a vehicle of 0.5% HPMC and 0.1% Tween 80 and stored under ambient conditions.

[0245] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were housed in a specially sourced pathogen-free (SPF) environment and allowed to acclimate to the new environment for at least three days before the start of any experiment. The mice were 6-8 weeks old at the time of transplantation.

[0246] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Animal Care Committee (IACUC). The animal facilities and programs operated under the standards of the Guidelines for the Management and Use of Laboratory Animals (NRC, 2011) and were accredited by the International Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0247] Preparation of PDX models The LUN2355-128-33 model was established for preclinical efficacy studies at GenenDesign (Shanghai, China). This PDX model was derived from a 49-year-old male Chinese NSCLC patient. EGFR mutations in the PDX model were confirmed by whole exom sequencing and PCR sequencing. Tumor fragments taken from the PDX model were subcutaneously transplanted into the right flank of female Balb / c nude mice. Mice were anesthetized with isoflurane and anesthesia was maintained throughout the transplantation surgery. The skin of the mice was cleaned with appropriate surgical scrubs and alcohol on the right flank. Aseptic surgery was used. A small skin incision was made using the sharp end of a trocar, and a 1.5 cm subcutaneous pocket along the right chest wall was formed by blunt incision with a 10-12 g trocar needle stylet. Tumor fragments (15-30 mm) 3 The trocar needle was inserted and advanced into the subcutaneous pocket in the right flank. The trocar incision was closed with sutures or wound clips, which were removed one week after closure.

[0248] Tumor size is 150-250 mm in volume 3 When the target was reached, mice with tumors were randomly divided into test groups, with 8 mice in each group. The randomization day is denoted as treatment day 0.

[0249] treatment Treatment was initiated on the same day as randomization. The treatment initiation day was denoted as treatment day 0. Mice were administered orally by vehicle control solution, compound I of formula 1 alone at 5 mg / kg BID, compound I of formula 1 alone at 15 mg / kg QD, and osimertinib alone at 15 mg / kg QD. Two additional groups received combination therapy with compound I of formula 1 and osimertinib; one group received the combination of compound I of formula 1 at 5 mg / kg BID and osimertinib at 15 mg / kg QD, and the other group received the combination of compound I of formula 1 at 15 mg / kg QD and osimertinib at 15 mg / kg QD. The dose was 5 mL / kg for each compound, and the interval between BID regimens was 8 hours. Osimertinib was administered 1 hour after administration of compound I QD, or after the first dose of the BID regimen in the combination groups.

[0250] Results Figure 12 shows graphs of tumor volume over the treatment period with regimens of the compound of formula I alone, osimertinib alone, and a combination of the compound of formula I and osimertinib in the EGFR L858R / T790M mutant NSCLC PDX model LUN2355-128-33. No significant change in body weight was observed in the control group and the treatment groups.

[0251] Conclusion As shown in Figure 12, the combination of the compound of formula I and osimertinib showed superior inhibition of tumor growth in the EGFR L858R / T790M mutant NSCLC PDX model LUN2355-128-33 compared to treatment with the compound of formula I alone or osimertinib alone.

[0252] Example 11 - Combination Therapy of the Compound of Formula I and Osimertinib in the EGFRexl9del Mutant CDX Model HCC827 / ER1 Materials Vehicle / control, 100 mM acetic acid in deionized water, adjusted to pH 4.8 - 5.0, was prepared and stored under ambient conditions during the 28-day administration period in mice.

[0253] The test article of the compound of formula I was freshly prepared weekly in a vehicle of 100 mM acetic acid buffer and stored under ambient conditions. The combination drug osimertinib was prepared weekly in a vehicle of 0.5% HPMC and 0. = 1% Tween 80 and stored under ambient conditions.

[0254] Female Balb / c nude mice were purchased from SPF(Beijing)Laboratory Animal Technology Co,Ltd. The mice were 9 - 11 weeks old at the time of transplantation. The mice were housed in a special pathogen-free (SPF) environmental breeding facility and acclimated to their new environment for at least 3 days before the start of any experiment.

[0255] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with guidelines approved by the Animal Care Committee (IACUC) of Crown Bioscience (Beijing, China). During the study, animal care and use were carried out in accordance with the regulations of the International Association for Accreditation and Evaluation of Laboratory Animal Care (AAALAC). In addition, all parts of this study conducted at Crown Bioscience (Beijing, China) adhered to research protocols that were made and approved by the principal investigator and applicable standard operating procedures (SOPs).

[0256] Preparation of xenograft models Human lung cancer HCC827 cells were purchased from ATCC. EGFR exon 19 deletion mutant and erlotinib-resistant MET-amplified HCC827 / ER1 cell line (Crown Bioscience UK) was induced from Crown Bioscience's HCC827 (ATCC) by culturing cells in the presence of escalating concentrations of erlotinib. HCC827 / ER1 cells were cultured at 37°C in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and 42 pM erlotinib, under air with 5% CO2. The medium was conventionally refreshed, and tumor cells were subcultured every 3-5 days at 80% confluence with trypsin-EDTA. Cells growing in the exponential growth phase were collected and counted for inoculation. Briefly, 5 × 10⁶ cells were mixed with 50% Matrigel. 6 A 100 μL cell suspension containing HCC827 / ER1 tumor cells was subcutaneously transplanted into the right flank of mice using a syringe. The health of the animals and tumor growth were monitored daily after transplantation. Tumor volume was measured twice weekly by caliper if the xenografted tumor was palpable and measurable.

[0257] The average tumor size is approximately 145 mm. 3 (104~189mm 3 When the tumor reached a certain level, the mice with tumors were randomly divided into test groups of 8 mice each. The randomization day is denoted as treatment day 0.

[0258] treatment Treatment was initiated on the same day as randomization. The treatment initiation day was denoted as treatment day 0. Mice were administered orally with a vehicle control solution, compound I alone at 30 mg / kg QD, and osimertinib alone at 15 mg / kg QD. Based on previously performed tolerability studies to identify the optimal dose of compound I in combination with osimertinib, the dose level of compound I was reduced from 30 mg / kg QD to 15 mg / kg QD for combination therapy. One additional group accepted combination treatment with compound I and osimertinib, receiving 15 mg / kg QD of compound I and 15 mg / kg QD of osimertinib. The dose was 5 mL / kg for each compound. In the combination group, osimertinib was administered 1 hour after the administration of compound I QD. This study was terminated on treatment day 28 as defined in the study protocol.

[0259] result Figure 13 shows graphs of tumor volume over treatment periods with regimens of compound I alone, osimertinib alone, and a combination of compound I and osimertinib in the EGFRex19del mutant CDX model HCC827 / ER1. No significant weight changes were observed in the control or treatment groups.

[0260] conclusion As shown in Figure 13, the combination of the compound of formula I with osimertinib demonstrated superior tumor growth inhibition in the EGFRex19del mutant CDX model HCC827 / ER1 compared to treatment with the compound of formula I alone or osimertinib alone.

[0261] Example 12 - Combination therapy of compound I and cetuximab in RAS / RAF wild-type PDX type CRC049 material Acetic acid in 100 mM water was prepared for the vehicle / control, with the pH adjusted to 4.8-5.0, and stored under ambient conditions for 28 days during administration to mice.

[0262] The test samples of the compound of formula I were freshly prepared weekly in a vehicle of 100 mM acetate buffer and stored under ambient conditions. The concomitant agent cetuximab was prepared in PBS and stored at 2–8°C.

[0263] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were housed in a specially sourced pathogen-free (SPF) environment and allowed to acclimate to the new environment for at least three days before the start of any experiment. The mice were 6-8 weeks old at the time of transplantation.

[0264] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with protocols and guidelines approved by GenenDesign's Animal Care Committee (IACUC). The animal facilities and programs operated under the standards of the Guidelines for the Management and Use of Laboratory Animals (NRC, 2011) and were accredited by the International Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Specifically, all parts of this study conducted at GenenDesign adhered to research protocols reviewed and approved by the IACUC and applicable standard operating procedures (SOPs).

[0265] Preparation of PDX models A CRC049 PDX model was established for efficacy studies using GenenDesign. This PDX model was derived from a 79-year-old male Chinese CRC patient. KRAS, NRAS, and HRAS, ARAF, BRAF, and RAFI genes in the PDX model were analyzed by whole exome sequencing and PCR sequencing. Tumor fragments taken from the PDX mouse model were subcutaneously transplanted into the right flank of female Balb / c nude mice. Mice were anesthetized with isoflurane and anesthesia was maintained throughout the transplantation surgery. The skin of the mice was cleaned with appropriate surgical scrub and alcohol on the right flank. Aseptic surgical procedures were used for transplantation. A small skin incision was made using the sharp end of a trocar, and a 1.5 cm subcutaneous pocket along the right chest wall was formed by blunt incision with a 10-12 g trocar needle stylet. Tumor fragment (15-30 mm) 3 The tumor was inserted into a trocar needle and advanced into a subcutaneous pocket in the right flank. The trocar incision was closed with sutures or wound clips and removed one week after closure. The tumor size was 143-249 mm in volume. 3 When the target was reached, mice with tumors were randomly divided into test groups, with 8 mice in each group. The randomization day is denoted as treatment day 0.

[0266] treatment Treatment was initiated on the same day as randomization. The treatment initiation day was denoted as treatment day 0. Mice were administered orally by vehicle control solution, cetuximab alone at 20 mg / kg BIW, and the compound of formula I alone at 30 mg / kg QD. One additional group accepted combination treatment with the compound of formula I and cetuximab, receiving the compound of formula I at 30 mg / kg QD and cetuximab at 20 mg / kg BIW. The dose was 5 mL / kg for each compound. In the combination group, cetuximab was administered 1 hour after the administration of the compound of formula I at 30 mg / kg QD. This study was concluded on day 28 as defined in the study protocol.

[0267] result Figure 14 shows graphs of tumor volume over treatment periods for regimens of compound I alone, cetuximab alone, and a combination of compound I and cetuximab in the RAS / RAF wild-type PDX model CRC049. No significant weight changes were observed in the control or treatment groups.

[0268] conclusion As shown in Figure 14, the combination of compound I and cetuximab demonstrated superior tumor growth inhibition in the RAS / RAF wild-type PDX model CRC049 compared to treatment with compound I alone or cetuximab alone.

[0269] Example 13 - Combination therapy of compound I and cetuximab in a RAS / RAF wild-type HPV-negative CDX model FaDu material Acetic acid in 100 mM water was prepared for the vehicle / control, with the pH adjusted to 4.8-5.0, and stored under ambient conditions for 28 days during administration to mice.

[0270] The test substance of formula I was freshly prepared weekly in a vehicle of 100 mM acetate buffer and stored under ambient conditions. (5 mg / mL, stored at 2-8°C) The concomitant agent cetuximab was diluted to 3 mg / mL with physiological saline before each administration.

[0271] Female Balb / c nude mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were 6-8 weeks old at the time of transplantation. The mice were housed in a specially sourced pathogen-free (SPF) environment and allowed to acclimate to the new environment for at least 3 days before the start of any experiment.

[0272] All procedures related to the handling, care, and treatment of animals in this study were carried out in accordance with the approval and guidelines of the WuXiAppTec Animal Care Committee (IACUC). During the study, animal care and use were carried out in accordance with the regulations of the International Association for Accreditation and Evaluation of Laboratory Animal Care (AAALAC). In addition, all parts of this study conducted at WuXiAppTec adhered to the research protocol approved by the principal investigator and applicable standard operating procedures (SOPs).

[0273] Preparation of xenograft models FaDu was a RAS / RAF wild-type human head and neck squamous cell carcinoma (pharyngeal carcinoma) cell line. The FaDu cell line was purchased from the American Cell Culture and Cell Line Preservation Center (ATCC® HTB-43®). FaDu cells were cultured at 37°C in an atmosphere of 5% CO2 in the air in EMEM containing 10% fetal bovine serum (FBS) and 1% antibiotic-antifungal agent (AA). The medium was changed every 2-3 days, and tumor cells were routinely subcultured at 80-90% confluence using trypsin-EDTA. Cells growing in the exponential growth phase were collected and counted for inoculation.

[0274] FaDu tumor cells were subcutaneously transplanted into mice. 5 × 10 6 A 200 μL cell suspension containing tumor cells was subcutaneously transplanted into the right flank of mice using a syringe. The animals' health and tumor growth were monitored daily. Tumor volume was measured twice weekly using a caliper if the tumor was palpable and measurable. The average tumor volume was 153 mm². 3 (75~217mm 3 When the tumor reached a certain level, mice with tumors were randomized into different groups of 8 mice each. The randomization day is denoted as treatment day 0.

[0275] treatment Treatment began on the day after randomization. The treatment start day is denoted as Treatment Day 1. Mice were administered orally by vehicle control solution, cetuximab alone at 30 mg / kg Q3D, and compound I alone at 10 mg / kg BID. One additional group accepted combination treatment with compound I and cetuximab, receiving 10 mg / kg BID of compound I and 30 mg / kg Q3D of cetuximab. The doses were 5 mL / kg for compound I and 10 mL / kg for cetuximab. The interval between BID regimens for compound I was 8 hours. In the combination group, cetuximab was administered 1 hour after the first dose of the compound I BID. This study was completed on day 28 as defined in the study protocol.

[0276] result Figure 15 shows graphs of tumor volume over treatment periods with regimens of compound I alone, cetuximab alone, and a combination of compound I and cetuximab in the RAS / RAF wild-type HPV-negative HNSCC CDX model FaDu. No significant weight changes were observed in the control or treatment groups.

[0277] conclusion As shown in Figure 15, the combination of compound I and cetuximab demonstrated superior tumor growth inhibition in the RAS / RAF wild-type HPV-negative HNSCC CDX model FaDu compared to treatment with compound I alone or cetuximab alone.

[0278] While the embodiments described above are explained in some detail by illustration and examples for the purpose of clarifying understanding, those skilled in the art will understand that certain changes and modifications may be practiced within the scope of the appended claims. Furthermore, each reference provided herein is incorporated by reference to the same extent that each individual reference is incorporated by reference. In the event of any conflict between this application and the references provided herein, this application shall prevail.

Claims

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

2. The composition described in claim 1, wherein EGFR in the subject is constitutively expressed.

3. 2. The composition of claim 1, wherein the cancer comprises an EGFR mutation selected from EGFR gene copy number gain, EGFR gene amplification, chromosome 7 polychromosis, EGFR L858R, EGFR exon 19 deletion, EGFR exon 19 insertion, EGFR L861Q, EGFR G719C, EGFR G719S, EGFR G719A, EGFR V765A, EGFR T783A, EGFR exon 20 insertion, EGFR splice variant, EGFR A289D, EGFR A289T, EGFR A289V, EGF R G598A, EGFR G598V, EGFR T790M, and EGFR C797S.

4. The composition of claim 1, wherein the EGFR inhibitor is selected from osimertinib, dacomitinib, lazertinib, nazartinib, neratinib, mobocertinib, afatinib, erlotinib, gefitinib, lapatinib, lifirafenib, amivantamab, cetuximab, panitumumab, necitumumab, mizotamab-cleztoclax, nimotuzumab and vandetanib.

5. The composition of claim 1, wherein the method comprises administering a third MAPK pathway inhibitor.

6. The composition of claim 1, wherein the dosage of the compound of formula I ranges from 20 mg to 400 mg per day.

7. The composition of claim 1, wherein the compound of formula I, or a pharma- ceutically acceptable salt thereof, is administered at a dosage of between about 20 mg and about 260 mg per day.

8. The composition of claim 1, wherein the compound of formula I, or a pharma- ceutically acceptable salt thereof, is administered at a dosage of between about 20 mg and about 60 mg per day.

9. The composition of claim 1, wherein the compound of formula I, or a pharma- ceutically acceptable salt thereof, is administered in a dosage of about 40 mg per day, or about 60 mg per day.

10. The composition of claim 1, wherein the compound of formula I, or a pharma- ceutically acceptable salt thereof, is administered twice daily at a dosage of about 10 mg to about 100 mg.

11. The composition of claim 1, wherein the compound of formula I, or a pharma- ceutically acceptable salt thereof, is administered twice daily at a dosage of about 20 mg to about 80 mg.

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

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

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

15. A kit comprising: (i) Formula I: [Case 2] or a pharma- ceutically acceptable salt thereof, (ii) EGFR inhibitor Including the kit.

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