Method of treating cancers with alkyne substituted quinazoline derivatives

Abstract

The present disclosure relates to methods of treating or preventing cancer (e.g., NSCLC), comprising administering Compound No. 1 or a pharmaceutically acceptable salt thereof, to a subject.

Description

Attorney Docket No.43701-02302 (ASET-057 / 001WO) METHOD OF TREATING CANCERS WITH ALKYNE SUBSTITUTED QUINAZOLINE DERIVATIVES RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Application No. 63 / 697,930, filed on September 23, 2024, which is incorporated by reference herein in its entirety. INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING

[0002] The Sequence Listing XML associated with this application is provided electronically in XML file format and is hereby incorporated by reference into the specification. The name of the XML file containing the Sequence Listing XML is “ASET- 057_001WO_ST26_SeqList.xml”. The XML file is 3,182 bytes, created on September 22, 2025, and is being submitted electronically. BACKGROUND

[0003] Mutations affecting either the intracellular catalytic domain or extracellular ligand binding domain of an ErbB receptor can generate oncogenic activity (the ErbB protein family consists of 4 members including ErbB-1, also named epidermal growth factor receptor (EGFR) and Erb-2, also named HER2 in humans). ErbB inhibitors are a known treatment for a number of cancers. However, not every patient is responsive satisfactorily to this treatment. Thus, there is a long-felt need in the art for new therapies that are able to address the variable responsiveness of cancer patients to known therapies. The present disclosure provides compositions and methods for preventing or treating cancer in patients with these oncogenic mutations without the variable responsiveness observed when patients having these ErbB mutants are treated using the existing standard of care. SUMMARY

[0004] In some aspects, the present disclosure provides a method of treating or preventing cancer (e.g., NSCLC) in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1: 1 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)(Compound No.1), or a pharmaceutically acceptable salt thereof.

[0005] In some aspects, the present disclosure provides Compound No.1, or a pharmaceutically acceptable salt thereof, for treating or preventing cancer (e.g., NSCLC) in a subject.

[0006] In some aspects, the present disclosure provides use of Compound No.1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating or preventing cancer (e.g., NSCLC) in a subject.

[0007] In some aspects, the present disclosure provides a pharmaceutical composition for treating or preventing cancer (e.g., NSCLC), comprising Compound No.1, or a pharmaceutically acceptable salt thereof.

[0008] In some aspects, the present disclosure provides a pharmaceutical kit for treating or preventing cancer (e.g., NSCLC), comprising Compound No.1, or a pharmaceutically acceptable salt thereof.

[0009] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the specification, the singular forms also include the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference. The references cited herein are not admitted to be prior art to the claimed invention. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods and examples are illustrative only and are not intended to be limiting. In the case of conflict between the chemical structures and names of the compounds disclosed herein, the chemical structures will control.

[0010] Other features and advantages of the disclosure will be apparent from the following detailed description and claims. 2 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) BRIEF DESCRIPTIONS OF FIGURES

[0011] FIG.1 is a graph showing exemplary EGFR mutations found in patients who had received prior EGFR TKI therapy.

[0012] FIG.2 is a graph showing the treatment related adverse events (TRAE) found in greater than 10% patients in a Phase 2 study after once daily administration of 200 mg of Compound No.1A. Rash includes rash, rash maculo-papular, rash pustular, and dermatitis acneiform.

[0013] FIG.3 is a graph showing preliminary response rate and durability in a Phase 2 study of Compound No.1A (200 mg once daily) in patients with EGFR-mutated non-small cell lung cancer. The graph shows for each patient the best response achieved, the sum target lesion (mm), the best percent reduction in the sum of diameters (%SoD), the EGFR mutations, prior first-line treatment and duration, prior second-line treatment and duration if applicable, and any non-EGFR pathway deregulation if applicable. “O” = osimertinib; “A” = afatinib; “C” = carboplatin; “Cis” = cisplatin; “Pem” = pemetrexed; “Pac” = paclitaxel; “B” = bevacizumab; “HER3-Dxd” = patritumab deruxtecan; “C1” = Cohort 1 (NCM); “C2” = Cohort 2 (C797S); “PD” = progressive disease; “SD” = stable disease; “uPR” = unconfirmed partial response; “cPR” = confirmed partial response; “uCR” = unconfirmed complete response; NCM = non-classical mutations.

[0014] FIG.4 is a graph showing preliminary response rate, durability, and duration of treatment in a Phase 2 study of Compound No.1A (200 mg once daily) in patients with EGFR-mutated non-small cell lung cancer. “Osi” = osimertinib; “CPI” = checkpoint inhibitor; “C” = carboplatin; “pem” = pemetrexed; “bev” = bevacizumab; “pac” = paclitaxel; “HER3-DXd” = patritumab deruxtecan; “PD” = progressive disease; “SD” = stable disease; “PR” = partial response; “uPR” = unconfirmed partial response; “cPR” = confirmed partial response; “uCR” = unconfirmed complete response.

[0015] FIGs.5A-5D depict the results of eight months of treatment with 200 mg Compound No.1A administered once daily in an exemplary patient with EGFR-mutated non-small cell lung cancer. The patient has two non-classical mutations (G719S and S768I) and previously received eight months of osimertinib treatment (first line) and two months of carboplatin and pemetrexed combination treatment (second line). FIG.5A is a graph showing the percent change in sum of diameters of an omentum lesion over eight months of treatment. FIG.5B is a chart showing the presence or absence of five other non-target lesions (pleura, LN-pleura, mesentery, and two brain lesions) which were monitored in addition to the omentum lesion 3 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) over the course of treatment, and observed to be absent by Month 8 of treatment. FIG.5C is a picture showing a scan of the omentum prior to Compound No.1A treatment. FIG.5D is a picture showing a scan of the omentum at Day 1 of Cycle 7 of Compound No.1A treatment. Each cycle was 21 days. “PR” = partial response; “CR” = complete response; “LN” = lymph node; “C7D1” = Day 1 of Cycle 7.

[0016] FIGs.6A-6C are graphs showing change in variant allele frequency percent (%VAF) and circulating tumor DNA percent (%ctDNA) clearance in three patients following 200 mg once daily treatment with Compound No.1A. FIG.6A depicts results from a patient with EGFR-Ex19del+C797S mutations with prior osimertinib treatment for 38.3 months. FIG.6B depicts results from a patient with EGFR-L858R+L833V+C797S mutations with prior osimertinib treatment for 50.0 months followed by osimertinib, carboplatin, pemetrexed, and bevacizumab combination treatment for 26.6 months. FIG.6C depicts results from a patient with EGFR-L858R+Y1016C+C797S mutations with prior carboplatin and paclitaxel combination treatment for 1.3 months followed by osimertinib treatment for 16.8 months. “PR” = partial response; “C1D1” = Day 1 of Cycle 1; “C3D1” = Day 1 of Cycle 3. Each cycle was 21 days. DETAILED DESCRIPTION

[0017] It is understood that the term “Compound No.1,” as used herein, refers to (E)-N-(4- ((3-chloro-2-fluorophenyl)amino)-7-((3-methyl-3-azabicyclo[3.1.0]hexan-1- yl)ethynyl)quinazolin-6-yl)-4-morpholinobut-2-enamide, which is a compound having the following structure:(Compound No.1).

[0018] It is understood that the term “Compound No.1A,” as used herein, refers to a compound having the following structure: 4 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)(Compound No.1A).

[0019] It is understood that the term “Compound No.1B,” as used herein, refers to a compound having the following structure:(Compound No.1B).

[0020] Compound Nos.1, 1A and 1B may be synthesized using methods described in PCT Publication No. WO2021 / 030711, the contents of which are incorporated in their entirety for all purposes. Methods and Uses of the Present Disclosure

[0021] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject Compound No.1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0022] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject Compound No.1 (e.g., Compound No.1A or Compound No.1B), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0023] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0024] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject a pharmaceutically effective 5 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) amount of Compound No.1 (e.g., Compound No.1A or Compound No.1B), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0025] In some aspects, the present disclosure provides Compound No.1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, for treating or preventing cancer in a subject.

[0026] In some aspects, the present disclosure provides Compound No.1 (e.g., Compound No.1A or Compound No.1B), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, for treating or preventing cancer in a subject.

[0027] In some aspects, the present disclosure provides use of Compound No.1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for treating or preventing cancer in a subject.

[0028] In some aspects, the present disclosure provides use of Compound No.1 (e.g., Compound No.1A or Compound No.1B), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for treating or preventing cancer in a subject.

[0029] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject Compound No.1, or a pharmaceutically acceptable salt thereof.

[0030] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof.

[0031] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1, or a pharmaceutically acceptable salt thereof.

[0032] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof.

[0033] In some aspects, the present disclosure provides Compound No.1, or a pharmaceutically acceptable salt thereof, for treating or preventing cancer in a subject.

[0034] In some aspects, the present disclosure provides Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof, for treating or preventing cancer in a subject. 6 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0035] In some aspects, the present disclosure provides use of Compound No.1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating or preventing cancer in a subject.

[0036] In some aspects, the present disclosure provides use of Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating or preventing cancer in a subject.

[0037] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject Compound No.1, or a pharmaceutically acceptable salt thereof.

[0038] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof.

[0039] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1, or a pharmaceutically acceptable salt thereof.

[0040] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof.

[0041] In some aspects, the present disclosure provides Compound No.1, or a pharmaceutically acceptable salt thereof, for treating cancer in a subject.

[0042] In some aspects, the present disclosure provides Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof, for treating cancer in a subject.

[0043] In some aspects, the present disclosure provides use of Compound No.1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating cancer in a subject.

[0044] In some aspects, the present disclosure provides use of Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating cancer in a subject.

[0045] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject Compound No.1A, or a pharmaceutically acceptable salt thereof. 7 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0046] In some aspects, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1A, or a pharmaceutically acceptable salt thereof.

[0047] In some aspects, the present disclosure provides Compound No.1A, or a pharmaceutically acceptable salt thereof, for treating or preventing cancer in a subject.

[0048] In some aspects, the present disclosure provides use of Compound No.1A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating or preventing cancer in a subject.

[0049] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject Compound No.1A, or a pharmaceutically acceptable salt thereof.

[0050] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1A, or a pharmaceutically acceptable salt thereof.

[0051] In some aspects, the present disclosure provides Compound No.1A, or a pharmaceutically acceptable salt thereof, for treating cancer in a subject.

[0052] In some aspects, the present disclosure provides use of Compound No.1A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating cancer in a subject.

[0053] In some embodiments, Compound No.1A, Compound No.1B, or a pharmaceutically acceptable salt of any one of the foregoing, is administered.

[0054] In some embodiments, Compound No.1A, or a pharmaceutically acceptable salt thereof, is administered.

[0055] In some embodiments, Compound No.1B, or a pharmaceutically acceptable salt thereof, is administered. Subjects and Diseases

[0056] In some embodiments, the subject is a mammal.

[0057] In some embodiments, the subject is a human.

[0058] In some embodiments, the subject is a human having an age of 18 years or older.

[0059] In some embodiments, the cancer is non-small cell lung cancer (NSCLC) or any subtype thereof.

[0060] In some embodiments, the cancer is NSCLC.

[0061] In some embodiments, the cancer is metastatic NSCLC. 8 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0062] In some embodiments, the cancer is recurrent NSCLC.

[0063] In some embodiments, the cancer is relapsed NSCLC.

[0064] In some embodiments, the cancer is refractory NSCLC.

[0065] In some embodiments, the cancer is advanced NSCLC.

[0066] In some embodiments, the cancer is locally advanced NSCLC.

[0067] In some embodiments, the NSCLC (e.g., the metastatic, recurrent relapsed, advanced, and / or locally advanced NSCLC) is characterized by at least one oncogenic mutation in the EGFR gene.

[0068] It is understood that a cancer that is characterized by at least one oncogenic mutation in a specific gene includes, but is not limited to, cancers whose primary oncogenic activity is thought to be driven by the at least one oncogenic mutation in that gene. In some embodiments, the oncogenic activity is driven by a combination of mutations within a gene.

[0069] In some embodiments, the NSCLC (e.g., the metastatic, recurrent relapsed, advanced, and / or locally advanced NSCLC) is characterized by an oncogenic variant of the EGFR protein.

[0070] It is understood that a cancer that is characterized by least one oncogenic variant of a protein is a cancer which includes, but is not limited to, cancers whose primary oncogenic activity is thought to be driven by the at least one oncogenic variant of the protein.

[0071] In some embodiments, the cancer is metastatic. In some embodiments, the cancer has metastasized to the brain. In some embodiments, the cancer has metastasized to the pleura. In some embodiments, the cancer has metastasized to the pleural lymph node. In some embodiments, the cancer has metastasized to the mesentery.

[0072] In some embodiments, the subject has at least one oncogenic mutation in the EGFR gene. In some embodiments, the subject has at least two oncogenic mutations in the EGFR gene. In some embodiments, the subject has at least three oncogenic mutations in the EGFR gene. In some embodiments, when a subject has at least two oncogenic mutations in the EGFR gene, the at least two oncogenic mutations occur within the same copy of the EGFR gene (i.e., the protein product of the EGFR gene will have the at least two oncogenic mutations). In some embodiments, when a subject has at least two oncogenic mutations in the EGFR gene, the at least two oncogenic mutations occur on different copies of the EGFR gene (i.e., there will be at least one protein product with one of the at least two oncogenic mutations and another protein product with the other oncogenic mutation of the at least two oncogenic mutations). 9 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0073] In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an oncogenic variant of the EGFR protein. In some embodiments, an oncogenic variant of the EGFR protein comprises one or more oncogenic mutations. In some embodiments, an oncogenic variant of the EGFR protein comprises at least one oncogenic mutation. In some embodiments, an oncogenic variant of the EGFR protein comprises at least two oncogenic mutations. In some embodiments, an oncogenic variant of the EGFR protein comprises at least three oncogenic mutations. In some embodiments, the subject has at least one cell that expresses a first oncogenic variant of the EGFR protein and a second oncogenic variant of the EGFR protein, wherein the first oncogenic variant of the EGFR protein and the second oncogenic of the EGFR protein are distinct.

[0074] As would be appreciated by the skilled artisan, in the context of a gene an oncogenic mutation can include, but is not limited to, a mutation that results in the substitution of one amino acid for another at a specific position within the protein product of the gene, a mutation that results in the substitution of one or more amino acids for one or more amino acids between two specific positions within protein product of the gene, a mutation that results in an insertion of one or more amino acids between two positions within protein product of the gene, a mutation that results in the deletion of one more amino acids between two positions within protein product of the gene, and mutation that results in a fusion of protein product of the gene, or portion thereof, with another protein, or portion thereof, or any combination thereof. As would be appreciated by the skilled artisan, in the context of a gene, a mutation can include, but is not limited to, a missense mutation, a nonsynonymous mutation, an insertion of one or more nucleotides, a deletion of one or more nucleotides, an inversion and a deletion-insertion. As would be appreciated by the skilled artisan, in the context of a gene, the gene can have one or more type of mutations, including combinations of different types of oncogenic mutations.

[0075] As would be appreciated by the skilled artisan, in the context of a protein, an oncogenic mutation can include, but is not limited to, the substitution of one amino acid for another at a specific position within the protein, the substitution of one or more amino acids for one or more amino acids between two specific positions within the protein, an insertion of one or more amino acids between two positions within the protein, a deletion of one more amino acids between two positions within the protein, and a fusion of the protein, or portion thereof, with another protein, or portion thereof, or any combination thereof. As would be appreciated by the skilled artisan, in the context of a protein, the protein can have one or 10 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) more type of oncogenic mutations, including combinations of different types of oncogenic mutations.

[0076] In some embodiments, the oncogenic mutation in the EGFR gene is a non-classical EGFR mutation. EGFR mutations can be classified into four subgroups: classical EGFR mutations, T790M-like mutations, Exon 20 loop insertion mutations, or P-loop and α-C-helix compressing (PACC) mutations. As would be appreciated by the skilled artisan, classical EGFR mutations include EGFR-Δ19 (e.g., the deletion of residues E746-A750 of EGFR) and EGFR-L858R. Accordingly, non-classical EGFR mutations are oncogenic mutations that are not classical EGFR mutations. Examples of such non-classical mutations include, but are not limited to, those listed in Table 2. A non-classical EGFR mutation can be any non-classical EGFR mutation that is known in the art. Table 1. Exemplary Non-Classical EGFR Mutations11 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0077] In some embodiments, the subject has at least one non-classical mutation in the EGFR gene. In some embodiments, the subject has at least two non-classical mutations in the EGFR gene. In some embodiments, the subject has at least three non-classical mutations in the EGFR gene.

[0078] In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an oncogenic EGFR variant, wherein the oncogenic EGFR variant comprises at least one non-classical mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an oncogenic EGFR variant, wherein the oncogenic EGFR variant comprises at least two non-classical mutations.

[0079] In some embodiments, a subject that has at least one non-classical EGFR mutation also has at least one classical EGFR mutation, e.g., EGFR-Δ19 or EGFR- L858R.

[0080] In some embodiments, the subject has at least one non-classical mutation and at least one classical mutation in the EGFR gene.

[0081] In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an oncogenic EGFR variant that comprises at least one non-classical EGFR mutation and at least one classical EGFR mutation.

[0082] In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses a first oncogenic EGFR variant comprising at least one non-classical EGFR mutation and a second oncogenic variant of the EGFR protein comprising at least one classical EGFR mutation.

[0083] In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses a first oncogenic EGFR variant comprising at least one non-classical EGFR mutation and at least one tumor and / or cancerous cell that expresses a second oncogenic EGFR variant comprising a classical EGFR mutation.

[0084] In some embodiments, in addition to the non-classical EGFR mutations and / or classical EGFR mutations, the subject has an EGFR-C797S mutation.

[0085] In some embodiments, the subject has at least one classical EGFR mutation, e.g., EGFR-Δ19, EGFR-L858R, or EGFR-C797S. Accordingly, in some embodiments, a subject has at least one classical mutation and a C797S mutation in the EGFR gene. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an 12 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) oncogenic EGFR variant that comprises at least one classical EGFR mutation and the C797S mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses a first oncogenic EGFR variant comprising at least one classical EGFR mutation and a second oncogenic variant of the EGFR protein comprising the C797S mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses a first oncogenic EGFR variant comprising at least one classical EGFR mutation and at least one tumor and / or cancerous cell that expresses a second oncogenic EGFR variant comprising the C797S mutation.

[0086] In some embodiments, a subject has at least one non-classical EGFR mutation and the EGFR-C797S mutation. Accordingly, in some embodiments, a subject has at least one non-classical mutation and a C797S mutation in the EGFR gene. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an oncogenic EGFR variant that comprises at least one non-classical EGFR mutation and the C797S mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses a first oncogenic EGFR variant comprising at least one non-classical EGFR mutation and a second oncogenic variant of the EGFR protein comprising the C797S mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses a first oncogenic EGFR variant comprising at least one non-classical EGFR mutation and at least one tumor and / or cancerous cell that expresses a second oncogenic EGFR variant comprising the C797S mutation.

[0087] In some embodiments, the non-classical EGFR mutation is a PACC mutation (see Robichaux, J.P., Le, X., Vijayan, R.S.K. et al. Structure-based classification predicts drug response in EGFR-mutant NSCLC. Nature 597, 732–737 (2021)., which is incorporated herein in its entirety for all purposes). PACC mutations are a specific subset of non-classical EGFR mutations. Without wishing to be bound by theory, PACC mutations alter the orientation of the P-loop and / or α-C-helix such that the binding pocket is compressed. PACC mutations occur within EGFR exons 18-21. Nonlimiting examples of PACC mutations include, but are not limited to EGFR-G719X, EGFR-L747X, EGFR-S768I, EGFR-L792X, and EGFR-T854I. PACC mutations can be identified through a variety of different means known to one skilled in the art, including the use of in silico mutational modeling methods in which one or more residues in EGFR are mutated from the wild-type residue to a putative PACC mutation. This in silico model can then be subjected to further analyses such as molecular dynamics simulations to determine the effect of the putative PACC mutation on the structure of the P-loop and / or α-C-helix. 13 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0088] In some embodiments, the PACC mutation is selected from any of the PACC mutations (including combinations of mutations) listed in Table 2. A PACC mutation can be any PACC mutation that is known in the art, including, but not limited to those listed in PCT Publication No. WO2022165214A1, PCT Publication No. WO2022165219A1, PCT Publication No. WO2024061203A1, US Patent Publication No. US20240108623A1, and Robichaux et al. Nature (see above). All of the aforementioned publications are incorporated herein by reference in their entireties for all purposes. Table 2. PACC Mutations

[0089] An oncogenic variant of EGFR that comprises at least one PACC mutation is referred to herein as an EGFR-PACC variant. In some embodiments, the EGFR-PACC variant comprises any of the mutations listed in Table 2. EGFR-PACC variants can comprise a combination of two or more of the PACC mutations listed in Table 2. In some embodiments, an EGFR-PACC variant comprises at least two PACC mutations. In some embodiments, an EGFR-PACC variant comprises at least three PACC mutations.

[0090] In some embodiments, the subject has at least one PACC mutation in the EGFR gene. In some embodiments, the subject has at least two PACC mutations in the EGFR gene. In some embodiments, the subject has at least three PACC mutations in the EGFR gene.

[0091] In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an EGFR-PACC variant.

[0092] In some embodiments, the subject has at least one PACC mutation and at least one other non-PACC mutation in the EGFR gene. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an EGFR-PACC variant that comprises at least one PACC mutation and at least one other non-PACC mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an EGFR-PACC variant and an oncogenic variant of the EGFR protein, wherein the oncogenic variant of the EGFR protein comprises at least one non-PACC mutation. In some embodiments, the subject has at least one tumor and / or cancerous cell that expresses an EGFR-PACC variant that 14 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) comprises at least one PACC mutation and at least one tumor and / or cancerous cell that expresses an oncogenic variant of the EGFR protein, wherein the oncogenic variant of the EGFR protein comprises at least one non-PACC mutation.

[0093] In some embodiments, the non-PACC mutation is a classical EGFR mutation, e.g., EGFR-Δ19 or EGFR- L858R.

[0094] In some embodiments, the non-PACC mutation is a non-classical mutation that is not a PACC mutation, including any such non-classical mutation listed in Table 1.

[0095] In some embodiments, the non-PACC mutation is EGFR-C797S.

[0096] Based on the above, Table 3 puts forth exemplary combinations of EGFR mutations that could occur in the subjects to be treated in the methods of the present disclosure. Table 3. Exemplary EGFR mutation combinations

[0097] In some embodiments, the subject has received at least one previous anti-NSCLC therapy. In some embodiments, the subject has become resistant to treatment with the at least one previous anti-NSCLC therapy.

[0098] In some embodiments the subject has received at least two previous anti-NSCLC therapies. In some embodiments, the subject has become resistant to treatment with the at least two previous anti-NSCLC therapies. 15 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0099] In some embodiments, the previous anti-NSCLC therapy is a systemic therapy (“anti- NSCLC systemic therapy”).

[0100] In some embodiments, the previous anti-NSCLC therapy comprises an EGFR- targeting therapy. In some embodiments, the EGFR-targeting therapy is a tyrosine kinase inhibitor (TKI). In some embodiments, the TKI is gefitinib, erlotinib, afatinib, osimertinib, crizotinib, alectinib, ceritinib, dabrafenib, sapitinib, dacomitinib, canertinib, pelitinib, icotinib, olmutinib, AC0010, EAI045, WZ4002, WZ8040, WZ3146, CO-1686, or AZD9291.

[0101] In some embodiments, the previous anti-NSCLC therapy is gefitinib, erlotinib, afatinib, osimertinib, necitumumab, crizotinib, alectinib, ceritinib, dabrafenib, trametinib, sapitinib, dacomitinib, canertinib, pelitinib, icotinib, olmutinib, AC0010, EAI045, WZ4002, WZ8040, WZ3146, CO-1686, AZD9291, carboplatin, cisplatin, pemetrexed, paclitaxel, bevacizumab, or patritumab deruxtecan, or a combination of any one of the foregoing.

[0102] In some embodiments, the previous anti-NSCLC therapy is osimertinib.

[0103] In some embodiments, the previous anti-NSCLC therapy is afatinib.

[0104] In some embodiments, the previous anti-NSCLC therapy is patritumab deruxtecan.

[0105] In some embodiments, the previous anti-NSCLC therapy comprises a combination of osimertinib, cisplatin, and pemetrexed.

[0106] In some embodiments, the previous anti-NSCLC therapy comprises a combination of osimertinib and bevacizumab.

[0107] In some embodiments, the previous anti-NSCLC therapy comprises a combination of carboplatin and paclitaxel.

[0108] In some embodiments, the previous anti-NSCLC therapy comprises a combination of osimertinib, carboplatin, and pemetrexed.

[0109] In some embodiments, the previous anti-NSCLC therapy comprises a combination of osimertinib, carboplatin, and paclitaxel.

[0110] In some embodiments, the previous anti-NSCLC therapy comprises a combination of osimertinib, carboplatin, pemetrexed, and bevacizumab.

[0111] In some embodiments, the subject has received at least one (e.g., one or two) previous anti-NSCLC therapy, wherein the only EGFR TKI that has been administered to the subject has been osimertinib.

[0112] Any of the subject characteristics described above can be combined with any other subject characteristic described above (e.g., in some embodiments, the subject has at least one non-classical EGFR mutation and has received at least one previous anti-NSCLC therapy). 16 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0113] In some embodiments, the subject has received at least one previous anti-NSCLC therapy and has at least one non-classical EGFR mutation. In some embodiments, the at least one non-classical EGFR mutation comprises a PACC mutation. In some embodiments, the subject has received one previous anti-NSCLC therapy. In some embodiments, the subject has received two previous anti-NSCLC therapies.

[0114] In some embodiments, the subject has received at least one previous anti-NSCLC therapy and has an EGFR-C797S mutation. In some embodiments, the subject has received one previous anti-NSCLC therapy. In some embodiments, the subject has received two previous anti-NSCLC therapies. In some embodiments, the only EGFR TKI that has been administered to the subject has been osimertinib.

[0115] In some embodiments, the subject has locally advanced or metastatic NSCLC, wherein the subject has at least one PACC mutation in EGFR and / or the EGFR-C797S mutation, wherein the subject has received at least one anti-NSCLC systemic therapy, wherein optionally the at least one anti-NSCLC systemic therapy comprised at least one EGFR-targeting therapy.

[0116] In some embodiments, the subject has locally advanced or metastatic NSCLC, wherein the subject has two or more EGFR mutations, wherein the subject has received at least one anti-NSCLC systemic therapy, wherein optionally the at least one anti-NSCLC systemic therapy comprises at least one EGFR-targeting therapy.

[0117] In some embodiments, the therapeutically effective amount reduces a severity of a sign or symptom of the cancer.

[0118] In some embodiments, the sign of the cancer comprises a tumor grade and a reduction of the severity of the sign comprises a decrease of the tumor grade.

[0119] In some embodiments, the sign of the cancer comprises a tumor metastasis and a reduction of the severity of the sign comprises an elimination of the metastasis or a reduction in the rate or extent the metastasis.

[0120] In some embodiments, the sign of the cancer comprises a tumor volume and a reduction of the severity of the sign comprises an elimination of the tumor or a reduction in the volume.

[0121] In some embodiments, the symptom of the cancer comprises pain and a reduction of the severity of the sign comprises an elimination or a reduction in the pain.

[0122] In some embodiments, the therapeutically effective amount induces a period of remission. 17 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0123] In some embodiments, the administration of Compound No.1 induces a period of remission in the subject. Administrations of Compound No.1

[0124] In some embodiments, Compound No.1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is administered.

[0125] In some embodiments, Compound No.1, or a pharmaceutically acceptable salt thereof, is administered.

[0126] In some embodiments, Compound No.1 is administered.

[0127] In some embodiments, a pharmaceutically acceptable salt of Compound No.1 is administered.

[0128] In some embodiments, a stereoisomer of Compound No.1 is administered.

[0129] In some embodiments, Compound No.1A, or a pharmaceutically acceptable salt thereof, is administered.

[0130] In some embodiments, Compound No.1A is administered.

[0131] In some embodiments, a pharmaceutically acceptable salt of Compound No.1A is administered.

[0132] In some embodiments, Compound No.1B, or a pharmaceutically acceptable salt thereof, is administered.

[0133] In some embodiments, Compound No.1B is administered.

[0134] In some embodiments, a pharmaceutically acceptable salt of Compound No.1B is administered.

[0135] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered daily.

[0136] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered once daily, twice daily, or three or more times daily.

[0137] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered once daily.

[0138] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered twice daily.

[0139] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three or more times daily. 18 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0140] In some embodiments the dosage administered reflects the weight of the free base, that is, the amount administered does not factor in the weight of any counterion present in a salt.

[0141] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of from about 1 mg to about 1250 mg, from about 5 mg to about 1150 mg, from about 10 mg to about 1100 mg, from about 15 mg to about 1000 mg, from about 25 mg to about 900 mg, from about 50 mg to about 800 mg, from about 75 mg to about 700 mg, from about 100 mg to about 600 mg, from about 150 mg to about 500 mg, or from about 200 mg to about 400 mg.

[0142] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 15 mg, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 800 mg, or about 1000 mg.

[0143] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 15 mg.

[0144] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 25 mg.

[0145] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 50 mg.

[0146] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 75 mg.

[0147] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 100 mg.

[0148] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 150 mg. 19 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0149] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 200 mg.

[0150] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 300 mg.

[0151] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 400 mg.

[0152] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 500 mg.

[0153] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 600 mg.

[0154] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 800 mg.

[0155] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 1000 mg.

[0156] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 60 mg / kg, about 180 mg / kg, about 600 mg / kg, or about 1800 mg / kg.

[0157] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 60 mg / kg.

[0158] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 180 mg / kg.

[0159] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 600 mg / kg. 20 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0160] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 1800 mg / kg.

[0161] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 5 mg / kg, about 15 mg / kg, about 50 mg / kg, or about 150 mg / kg.

[0162] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 5 mg / kg.

[0163] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 15 mg / kg.

[0164] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 50 mg / kg.

[0165] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered at a dosage (e.g., a daily dosage) of about 150 mg / kg.

[0166] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered weekly.

[0167] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three times per week.

[0168] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three times per week (e.g., at Day 1, Day 3, and Day 5 of each week, wherein Day 1 is the first day of administration of Compound No.1).

[0169] It is understood that Day 1 can be any day of a calendar week.

[0170] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered orally.

[0171] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is orally administered three times per week. 21 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0172] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three times per week (e.g., at day 1, day 3, and day 5) at a dosage of about 300 mg, about 400 mg, or about 500 mg.

[0173] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three times per week (e.g., at day 1, day 3, and day 5) at a dosage of about 300 mg.

[0174] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three times per week (e.g., at day 1, day 3, and day 5) at a dosage of about 400 mg.

[0175] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered three times per week (e.g., at day 1, day 3, and day 5) at a dosage of about 500 mg.

[0176] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered for about 21 days, about 28 days, about 35 days, about 42 days, about 63 days, about 84 days, about 105 days, about 126 days, about 147 days, about 168 days, about 189 days, or about 210 days.

[0177] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered for longer than 210 days.

[0178] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered until a progression of cancer or an adverse event (e.g., an intolerable toxicity) is observed.

[0179] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered for about 3 weeks.

[0180] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered for about 21 days.

[0181] In some embodiments, Compound No.1 (e.g., Compound No.1A or Compound No. 1B), or a pharmaceutically acceptable salt thereof, is administered for about 21 days, followed by a 30-day drug holiday.

[0182] In some embodiments, the treating or preventing lasts about 1 month, about 2 months, about 3 months, about 6 months, about 9 months, about 12 months, about 15 months, about 18 months, about 21 months, or about 24 months.

[0183] In some embodiments, the treating or preventing comprises one or more treatment cycles, wherein each treatment cycle comprises administering Compound No.1 (e.g., 22 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof, for about 21 days, followed by a 30-day drug holiday.

[0184] In some embodiments, the treating lasts about 1 month, about 2 months, about 3 months, about 6 months, about 9 months, about 12 months, about 15 months, about 18 months, about 21 months, or about 24 months.

[0185] In some embodiments, the treating comprises one or more treatment cycles, wherein each treatment cycle comprises administering Compound No.1 (e.g., Compound No.1A or Compound No.1B), or a pharmaceutically acceptable salt thereof, for about 21 days, followed by a 30-day drug holiday Definitions

[0186] It will be understood that while compounds disclosed herein may be presented in one particular configuration. Such particular configuration is not to be construed as limiting the disclosure to one or another isomer, tautomer, regioisomer or stereoisomer, nor does it exclude mixtures of isomers, tautomers, regioisomers or stereoisomers. In some embodiments, the presentation of a compound herein in a particular configuration intends to encompass, and to refer to, each of the available isomers, tautomers, regioisomers, and stereoisomers of the compound, or any mixture thereof; while the presentation further intends to refer to the specific configuration of the compound.

[0187] Further, it will be understood that while compounds disclosed herein may be presented without specified configuration (e.g., without specified stereochemistry). Such presentation intends to encompass all available isomers, tautomers, regioisomers, and stereoisomers of the compound. In some embodiments, the presentation of a compound herein without specified configuration intends to refer to each of the available isomers, tautomers, regioisomers, and stereoisomers of the compound, or any mixture thereof.

[0188] As used herein, the term “isomerism” means compounds that have identical molecular formulae but differ in the sequence of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers.” Stereoisomers that are not mirror images of one another are termed “diastereoisomers,” and stereoisomers that are non-superimposable mirror images of each other are termed “enantiomers” or sometimes optical isomers. A mixture containing equal amounts of individual enantiomeric forms of opposite chirality is termed a “racemic mixture.” 23 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0189] As used herein, the term “chiral center” refers to a carbon atom bonded to four nonidentical substituents.

[0190] As used herein, the term “chiral isomer” means a compound with at least one chiral center. Compounds with more than one chiral center may exist either as an individual diastereomer or as a mixture of diastereomers, termed “diastereomeric mixture.” When one chiral center is present, a stereoisomer may be characterized by the absolute configuration (R or S) of that chiral center. Absolute configuration refers to the arrangement in space of the substituents attached to the chiral center. The substituents attached to the chiral center under consideration are ranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog. (Cahn et al., Angew. Chem. Inter. Edit.1966, 5, 385; errata 511; Cahn et al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc.1951 (London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J. Chem. Educ.1964, 41, 116).

[0191] As used herein, the term “geometric isomer” means the diastereomers that owe their existence to hindered rotation about double bonds or a cycloalkyl linker (e.g., 1,3- cyclobutyl). These configurations are differentiated in their names by the prefixes cis and trans, or Z and E, which indicate that the groups are on the same or opposite side of the double bond in the molecule according to the Cahn-Ingold-Prelog rules.

[0192] The terms “nucleic acid” and “polynucleotide” are used interchangeably herein to refer to single- or double-stranded RNA, DNA, or mixed polymers. Polynucleotides may include genomic sequences, extra-genomic and plasmid sequences, and smaller engineered gene segments that express, or may be adapted to express polypeptides.

[0193] An “isolated nucleic acid” is a nucleic acid that is substantially separated from other genome DNA sequences as well as proteins or complexes such as ribosomes and polymerases, which naturally accompany a native sequence. The term embraces a nucleic acid sequence that has been removed from its naturally occurring environment and includes recombinant or cloned DNA isolates and chemically synthesized analogues or analogues biologically synthesized by heterologous systems. A substantially pure nucleic acid includes isolated forms of the nucleic acid. Of course, this refers to the nucleic acid as originally isolated and does not exclude genes or sequences later added to the isolated nucleic acid by the hand of man.

[0194] The term “polypeptide” is used in its conventional meaning, i.e., as a sequence of amino acids. The polypeptides are not limited to a specific length of the product. Peptides, oligopeptides, and proteins are included within the definition of polypeptide, and such terms may be used interchangeably herein unless specifically indicated otherwise. This term also 24 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) does not refer to or exclude post-expression modifications of the polypeptide, for example, glycosylations, acetylations, phosphorylations and the like, as well as other modifications known in the art, both naturally occurring and non-naturally occurring. A polypeptide may be an entire protein, or a subsequence thereof.

[0195] An “isolated polypeptide” is one that has been identified and separated and / or recovered from a component of its natural environment. In preferred embodiments, the isolated polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or, preferably, silver stain. Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.

[0196] A “native sequence” polynucleotide is one that has the same nucleotide sequence as a polynucleotide derived from nature. A “native sequence” polypeptide is one that has the same amino acid sequence as a polypeptide (e.g., EGFR) derived from nature (e.g., from any species). Such native sequence polynucleotides and polypeptides can be isolated from nature or can be produced by recombinant or synthetic means.

[0197] A polynucleotide “variant,” as the term is used herein, is a polynucleotide that typically differs from a polynucleotide specifically disclosed herein in one or more substitutions, deletions, additions and / or insertions.

[0198] A polypeptide “variant,” as the term is used herein, is a polypeptide that typically differs from a polypeptide specifically disclosed herein in one or more substitutions, deletions, additions and / or insertions, or inversions. Such variants may be naturally occurring, non-naturally occurring, or may be synthetically generated.

[0199] EGFR mutations (or variants) of the disclosure may comprise one or more substitutions, deletions, additions and / or insertions, or inversions of the amino acid sequence that are alter the function of the resultant protein. Mutations may be detected, for example, by comparison or alignment of a nucleic or amino acid sequence with a wild-type sequence.

[0200] When comparing polynucleotide and polypeptide sequences, two sequences are said to be “identical” if the sequence of nucleotides or amino acids in the two sequences is the same when aligned for maximum correspondence, as described below. Comparisons between two sequences are typically performed by comparing the sequences over a comparison 25 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) window to identify and compare local regions of sequence similarity. A “comparison window” as used herein, refers to a segment of at least about 20 contiguous positions, usually 30 to about 75, 40 to about 50, in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned.

[0201] Optimal alignment of sequences for comparison may be conducted using the Megalign program in the Lasergene suite of bioinformatics software (DNASTAR, Inc., Madison, WI), using default parameters. This program embodies several alignment schemes described in the following references: Dayhoff, M.O. (1978) A model of evolutionary change in proteins – Matrices for detecting distant relationships. In Dayhoff, M.O. (ed.) Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, Washington DC Vol.5, Suppl.3, pp.345-358; Hein J. (1990) Unified Approach to Alignment and Phylogenes pp.626-645 Methods in Enzymology vol.183, Academic Press, Inc., San Diego, CA; Higgins, D.G. and Sharp, P.M. (1989) CABIOS 5:151-153; Myers, E.W. and Muller W. (1988) CABIOS 4:11-17; Robinson, E.D. (1971) Comb. Theor 11:105; Santou, N. Nes, M. (1987) Mol. Biol. Evol.4:406-425; Sneath, P.H.A. and Sokal, R.R. (1973) Numerical Taxonomy – the Principles and Practice of Numerical Taxonomy, Freeman Press, San Francisco, CA; Wilbur, W.J. and Lipman, D.J. (1983) Proc. Natl. Acad., Sci. USA 80:726- 730.

[0202] Alternatively, optimal alignment of sequences for comparison may be conducted by the local identity algorithm of Smith and Waterman (1981) Add. APL. Math 2:482, by the identity alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol.48:443, by the search for similarity methods of Pearson and Lipman (1988) Proc. Natl. Acad. Sci. USA 85: 2444, by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, WI), or by inspection.

[0203] One preferred example of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977) Nucl. Acids Res.25:3389-3402 and Altschul et al. (1990) J. Mol. Biol.215:403-410, respectively. BLAST and BLAST 2.0 can be used, for example with the parameters described herein, to determine percent sequence identity for the polynucleotides and polypeptides of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information.

[0204] In one illustrative example, cumulative scores can be calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N 26 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) (penalty score for mismatching residues; always <0). Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915) alignments, (B) of 50, expectation (E) of 10, M=5, N=-4 and a comparison of both strands.

[0205] For amino acid sequences, a scoring matrix can be used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment.

[0206] In one approach, the “percentage of sequence identity” is determined by comparing two optimally aligned sequences over a window of comparison of at least 20 positions, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) of 20 percent or less, usually 5 to 15 percent, or 10 to 12 percent, as compared to the reference sequences (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid bases or amino acid residues occur in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the reference sequence (i.e., the window size) and multiplying the results by 100 to yield the percentage of sequence identity.

[0207] A wild-type EGFR sequence of the disclosure may comprise or consist of the amino acid sequence of: 1 mrpsgtagaa llallaalcp asraleekkv cqgtsnkltq lgtfedhfls lqrmfnncev 61 vlgnleityv qrnydlsflk tiqevagyvl ialntverip lenlqiirgn myyensyala 121 vlsnydankt glkelpmrnl qeilhgavrf snnpalcnve siqwrdivss dflsnmsmdf 27 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) 181 qnhlgscqkc dpscpngscw gageencqkl tkiicaqqcs grcrgkspsd cchnqcaagc 241 tgpresdclv crkfrdeatc kdtcpplmly npttyqmdvn pegkysfgat cvkkcprnyv 301 vtdhgscvra cgadsyemee dgvrkckkce gpcrkvcngi gigefkdsls inatnikhfk 361 nctsisgdlh ilpvafrgds fthtppldpq eldilktvke itgflliqaw penrtdlhaf 421 enleiirgrt kqhgqfslav vslnitslgl rslkeisdgd viisgnknlc yantinwkkl 481 fgtsgqktki isnrgensck atgqvchalc spegcwgpep rdcvscrnvs rgrecvdkck 541 llegeprefv enseciqchp eclpqamnit ctgrgpdnci qcahyidgph cvktcpagvm 601 genntlvwky adaghvchlc hpnctygctg pglegcptng pkipsiatgm vgalllllvv 661 algiglfmrr rhivrkrtlr rllqerelve pltpsgeapn qallrilket efkkikvlgs 721 gafgtvykgl wipegekvki pvaikelrea tspkankeil deayvmasvd nphvcrllgi 781 cltstvqlit qlmpfgclld yvrehkdnig sqyllnwcvq iakgmnyled rrlvhrdlaa 841 rnvlvktpqh vkitdfglak llgaeekeyh aeggkvpikw malesilhri ythqsdvwsy 901 gvtvwelmtf gskpydgipa seissilekg erlpqppict idvymimvkc wmidadsrpk 961 freliiefsk mardpqrylv iqgdermhlp sptdsnfyra lmdeedmddv vdadeylipq 1021 qgffsspsts rtpllsslsa tsnnstvaci drnglqscpi kedsflqrys sdptgalted 1081 siddtflpvp eyinqsvpkr pagsvqnpvy hnqplnpaps rdphyqdphs tavgnpeyln 1141 tvqptcvnst fdspahwaqk gshqisldnp dyqqdffpke akpngifkgs taenaeylrv 1201 apqssefiga (SEQ ID NO: 1, corresponding to epidermal growth factor receptor [Homo sapiens] and Genbank Accession No. CAA25240). 28 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0208] Based on the definitions given throughout the application the skilled person knows which combinations are synthetically feasible and realistic, e.g., typically combinations of groups leading to heteroatoms directly linked to each other are not contemplated.

[0209] As used herein, the term “pharmaceutically acceptable salt” refers to a derivative of the compound of the present disclosure wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include, but are not limited to, those derived from inorganic and organic acids selected from 2-acetoxybenzoic, 2-hydroxyethane sulfonic, acetic, ascorbic, benzene sulfonic, benzoic, bicarbonic, carbonic, citric, edetic, ethane disulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic, gluconic, glutamic, glycolic, glycollyarsanilic, hexylresorcinic, hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxymaleic, hydroxynaphthoic, isethionic, lactic, lactobionic, lauryl sulfonic, maleic, malic, mandelic, methane sulfonic, napsylic, nitric, oxalic, pamoic, pantothenic, phenylacetic, phosphoric, polygalacturonic, propionic, salicylic, stearic, subacetic, succinic, sulfamic, sulfanilic, sulfuric, tannic, tartaric, toluene sulfonic, and the commonly occurring amine acids, e.g., glycine, alanine, phenylalanine, arginine, etc. Other examples of pharmaceutically acceptable salts include hexanoic acid, cyclopentane propionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonic acid, 2- naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo- [2.2.2]-oct-2-ene-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, muconic acid, and the like. The present disclosure also encompasses salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N- methylglucamine, and the like. In the salt form, it is understood that the ratio of the compound to the cation or anion of the salt can be 1:1, or any ratio other than 1:1, e.g., 3:1, 2:1, 1:2, or 1:3. It is to be understood that all references to pharmaceutically acceptable salts include solvent addition forms (solvates) or crystal forms (polymorphs) as defined herein, of the same salt. 29 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0210] It is understood that the compounds described herein include the compounds themselves, as well as their pharmaceutically acceptable salts, and their solvates, if applicable. A pharmaceutically acceptable salt, for example, can be formed between a pharmaceutically acceptable anion and a positively charged group (e.g., amino) on a compound. Suitable anions include chloride, bromide, iodide, sulfate, bisulfate, sulfamate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, glutamate, glucuronate, glutarate, malate, maleate, succinate, fumarate, tartrate, tosylate, salicylate, lactate, naphthalenesulfonate, and acetate (e.g., trifluoroacetate).

[0211] As used herein, the expressions “one or more of A, B, or C,” “one or more A, B, or C,” “one or more of A, B, and C,” “one or more A, B, and C,” “selected from the group consisting of A, B, and C”, “selected from A, B, and C”, and the like are used interchangeably and all refer to a selection from a group consisting of A, B, and / or C, i.e., one or more As, one or more Bs, one or more Cs, or any combination thereof, unless indicated otherwise.

[0212] In some embodiments, the term “about” refers to a range of values that fall within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). In some embodiments, the term “about” refers to a range of values that fall within 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0213] As used herein, the term “daily dosage” refers to the total dose of a compound of the present disclosure administered in a day.

[0214] As used herein, the term “daily dosage” refers to the total dose of a compound of the present disclosure administered in a 24 hour period.

[0215] It is understood that, throughout the description, where compositions are described as having, including, or comprising specific components, it is contemplated that compositions also consist essentially of, or consist of, the recited components. Similarly, where methods or processes are described as having, including, or comprising specific process steps, the processes also consist essentially of, or consist of, the recited processing steps. Further, it should be understood that the order of steps or order for performing certain actions is immaterial so long as the invention remains operable. Moreover, two or more steps or actions can be conducted simultaneously. 30 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0216] It is understood that compounds of the present disclosure can be prepared in a variety of ways using commercially available starting materials, compounds known in the literature, or from readily prepared intermediates, by employing standard synthetic methods and procedures either known to those skilled in the art, or which will be apparent to the skilled artisan in light of the teachings herein. Standard synthetic methods and procedures for the preparation of organic molecules and functional group transformations and manipulations can be obtained from the relevant scientific literature or from standard textbooks in the field. Although not limited to any one or several sources, classic texts such as Smith, M. B., March, J., March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5thedition, John Wiley & Sons: New York, 2001; Greene, T.W., Wuts, P.G. M., Protective Groups in Organic Synthesis, 3rdedition, John Wiley & Sons: New York, 1999; R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); L. Fieser and M. Fieser, Fieser and Fieser’s Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), incorporated by reference herein, are useful and recognized reference textbooks of organic synthesis known to those in the art.

[0217] As used herein, the term “subject” is interchangeable with the term “subject in need thereof,” both of which refer to a subject having a disease or having an increased risk of developing the disease. A “subject” includes a mammal. The mammal can be e.g., a human or appropriate non-human mammal, such as primate, mouse, rat, dog, cat, cow, horse, goat, camel, sheep or a pig. The subject can also be a bird or fowl. In some embodiments, the mammal is a human.

[0218] As used herein, the term “treating” or “treat” describes the management and care of a patient for the purpose of combating a disease, condition, or disorder and includes the administration of a compound of the present disclosure, or a pharmaceutically acceptable salt, polymorph or solvate thereof, to alleviate the symptoms or complications of a disease, condition or disorder, or to eliminate the disease, condition or disorder. The term “treat” can also include treatment of a cell in vitro or an animal model.

[0219] It is understood that a compound of the present disclosure, or a pharmaceutically acceptable salt, polymorph or solvate thereof, can or may also be used to prevent a relevant disease, condition or disorder, or used to identify suitable candidates for such purposes.

[0220] As used herein, the term “preventing,” “prevent,” or “protecting against” describes reducing or eliminating the onset of the symptoms or complications of such disease, condition or disorder. 31 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0221] As used herein, the term “pharmaceutically acceptable” refers to those compounds, anions, cations, materials, compositions, carriers, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0222] As used herein, the term “pharmaceutically effective amount,” refers to an amount of a pharmaceutical agent to treat, ameliorate, or prevent an identified disease or condition, or to exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend upon the subject’s body weight, size, and health; the nature and extent of the condition; and the therapeutic or combination of therapeutics selected for administration. Pharmaceutically effective amounts for a given situation can be determined by routine experimentation that is within the skill and judgment of the clinician.

[0223] As used herein, the term “pharmaceutically acceptable salts” refer to derivatives of the compounds of the present disclosure wherein the parent compound is modified by making acid or base salts thereof. In some embodiments, a pharmaceutically acceptable salt of a compound is also a prodrug of the compound. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include, but are not limited to, those derived from inorganic and organic acids selected from 2-acetoxybenzoic, 2- hydroxyethane sulfonic, acetic, ascorbic, benzene sulfonic, benzoic, bicarbonic, carbonic, citric, edetic, ethane disulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic, gluconic, glutamic, glycolic, glycollyarsanilic, hexylresorcinic, hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxymaleic, hydroxynaphthoic, isethionic, lactic, lactobionic, lauryl sulfonic, maleic, malic, mandelic, methane sulfonic, napsylic, nitric, oxalic, pamoic, pantothenic, phenylacetic, phosphoric, polygalacturonic, propionic, salicylic, stearic, subacetic, succinic, sulfamic, sulfanilic, sulfuric, tannic, tartaric, toluene sulfonic, and the commonly occurring amine acids, e.g., glycine, alanine, phenylalanine, arginine, etc.

[0224] Other examples of pharmaceutically acceptable salts include hexanoic acid, cyclopentane propionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic 32 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) acid, camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylic acid, 3- phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, muconic acid, and the like. The present disclosure also encompasses salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. In the salt form, it is understood that the ratio of the compound to the cation or anion of the salt can be 1:1, or any ration other than 1:1, e.g., 3:1, 2:1, 1:2, or 1:3.

[0225] As used herein, the term “prodrug” refers to any agent which, when administered to a mammal, is converted in whole or in part to a targeted compound. In some embodiments, the prodrug of a compound is also a pharmaceutically acceptable salt of the compound.

[0226] The compounds, or pharmaceutically acceptable salts thereof, are administered orally, nasally, transdermally, pulmonary, inhalationally, buccally, sublingually, intraperitoneally, subcutaneously, intramuscularly, intravenously, rectally, intrapleurally, intrathecally and parenterally. In one embodiment, the compound is administered orally. One skilled in the art will recognize the advantages of certain routes of administration.

[0227] The dosage regimen utilizing the compounds is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed.

[0228] Techniques for formulation and administration of the disclosed compounds of the disclosure can be found in Remington: the Science and Practice of Pharmacy, 19thedition, Mack Publishing Co., Easton, PA (1995). In an embodiment, the compounds described herein, and the pharmaceutically acceptable salts thereof, are used in pharmaceutical preparations in combination with one or more of a pharmaceutically acceptable adjuvant, carrier, excipient, or diluent. Suitable pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous or organic solutions. The compounds will be present in such pharmaceutical compositions in amounts sufficient to provide the desired dosage amount in the range described herein.

[0229] All percentages and ratios used herein, unless otherwise indicated, are by weight. Other features and advantages of the present disclosure are apparent from the different examples. The provided examples illustrate different components and methodology useful in practicing the present disclosure. The examples do not limit the claimed disclosure. Based on 33 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) the present disclosure the skilled artisan can identify and employ other components and methodology useful for practicing the present disclosure.

[0230] All publications and patent documents cited herein are incorporated herein by reference as if each such publication or document was specifically and individually indicated to be incorporated herein by reference. Citation of publications and patent documents is not intended as an admission that any is pertinent prior art, nor does it constitute any admission as to the contents or date of the same. The invention having now been described by way of written description, those of skill in the art will recognize that the invention can be practiced in a variety of embodiments and that the foregoing description and examples below are for purposes of illustration and not limitation of the claims that follow. Exemplary Embodiments

[0231] Exemplary Embodiment 1. A method of treating NSCLC in a subject, comprising administering to the subject a pharmaceutically effective amount of Compound No.1:or a pharmaceutically acceptable salt thereof.

[0232] Exemplary Embodiment 2. Compound No.1, or a pharmaceutically acceptable salt thereof, for use in treating NSCLC in a subject.

[0233] Exemplary Embodiment 3. Use of Compound No.1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating NSCLC in a subject.

[0234] Exemplary Embodiment 4. The method, compound for use, or use of any one of Exemplary Embodiments 1-3, wherein the NSCLC is metastatic NSCLC.

[0235] Exemplary Embodiment 5. The method, compound for use, or use of any one of Exemplary Embodiments 1-3, wherein the NSCLC is locally advanced NSCLC.

[0236] Exemplary Embodiment 6. The method, compound for use, or use of any one of Exemplary Embodiments 1-5, wherein the subject has not previously received an anti- NSCLC therapy and has at least one non-classical EGFR mutation. 34 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0237] Exemplary Embodiment 7. The method, compound for use, or use of any one of Exemplary Embodiments 1-5, wherein the subject has previously received at least one previous line of anti-NSCLC therapy, optionally wherein the at least one previous line of anti-NCLSC therapy was a systemic anti-NSCLC therapy, and wherein the subject has: i) at least one classical EGFR mutation and the EGFR-C797S mutation; ii) at least one classical EGFR mutation and at least one non-classical EGFR mutation; iii) at least one classical EGFR mutation, at least one non-classical EGFR mutation, and the EGFR-C797S mutation; or iv) the EGFR-C797S mutation.

[0238] Exemplary Embodiment 8. The method, compound for use, or use of Exemplary Embodiment 7, wherein the subject has received one previous line of anti-NSCLC therapy.

[0239] Exemplary Embodiment 9. The method, compound for use, or use of Exemplary Embodiment 7, wherein the subject has received two previous lines of anti-NCLSC therapy.

[0240] Exemplary Embodiment 10. The method, compound for use, or use of any one of Exemplary Embodiments 7-9, wherein the subject has received at least one previous line of anti-NCLSC therapy comprising at least one EGFR-targeting therapeutic, optionally wherein the EGFR-targeting therapeutic is a tyrosine kinase inhibitor (TKI), optionally wherein the TKI is osimertinib.

[0241] Exemplary Embodiment 11. The method, compound for use, or use of any one of the preceding Exemplary Embodiments, wherein the at least one non-classical EGFR mutation is selected from those put forth in Table 1.

[0242] Exemplary Embodiment 12. The method, compound for use, or use of any one of the preceding Exemplary Embodiments, wherein the at least one non-classical EGFR mutation comprises at least one P-loop and α-C-helix compressing (PACC) mutation, optionally wherein the PACC mutation is selected from those put forth in Table 2.

[0243] Exemplary Embodiment 13. The method, compound for use, or use of any one of the preceding Exemplary Embodiments, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered daily.

[0244] Exemplary Embodiment 14. The method, compound for use, or use of any one of the preceding Exemplary Embodiments, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered at a daily dosage of about 200 mg. 35 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0245] Exemplary Embodiment 15. The method, compound for use, or use of any one of the preceding Exemplary Embodiments, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered at a daily dosage of about 200 mg.

[0246] Exemplary Embodiment 16. The method, compound for use, or use of any one of Exemplary Embodiments 1-15, wherein Compound No.1 is administered.

[0247] Exemplary Embodiment 17. The method, compound for use, or use of any one of Exemplary Embodiments 1-15, wherein Compound No.1A:, or a pharmaceutically acceptable salt thereof, is administered.

[0248] Exemplary Embodiment 18. The method, compound for use, or use of any one of Exemplary Embodiments 1-15, wherein Compound No.1A is administered.

[0249] Exemplary Embodiment 19. The method, compound for use, or use of any one of Exemplary Embodiments 1-15, wherein Compound No.1B:, or a pharmaceutically acceptable salt thereof, is administered.

[0250] Exemplary Embodiment 20. The method, compound for use, or use of any one of Exemplary Embodiments 1-15, wherein Compound No.1B is administered.

[0251] Exemplary Embodiment A-1. A method of treating non-small cell lung cancer in a subject, comprising administering to the subject Compound No.1: 36 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)(Compound No.1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0252] Exemplary Embodiment A-2. Compound No.1:(Compound No.1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, for use in the treatment of non-small cell lung cancer in a subject.

[0253] Exemplary Embodiment A-3. Use of Compound No.1:(Compound No.1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the treatment of non- small cell lung cancer in a subject.

[0254] Exemplary Embodiment A-4. Use of Compound No.1: 37 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)(Compound No.1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for the treatment of non-small cell lung cancer in a subject.

[0255] Exemplary Embodiment A-5. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-4, wherein the non-small cell lung cancer is metastatic non-small cell lung cancer .

[0256] Exemplary Embodiment A-6. The method, compound for use, or use of Exemplary Embodiment 5, wherein the non-small cell lung cancer has metastasized to the brain.

[0257] Exemplary Embodiment A-7. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-4, wherein the non-small cell lung cancer is locally advanced non-small cell lung cancer.

[0258] Exemplary Embodiment A-8. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-7, wherein the subject has previously received at least one anti-non-small cell lung cancer therapy, wherein the subject has: i) at least one classical EGFR mutation and a EGFR-C797S mutation; ii) at least one classical EGFR mutation and at least one non-classical EGFR mutation; iii) at least one classical EGFR mutation, at least one non-classical EGFR mutation, and a EGFR-C797S mutation; or iv) a EGFR-C797S mutation.

[0259] Exemplary Embodiment A-9. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-8, wherein the subject has received one previous anti-non-small cell lung cancer therapy.

[0260] Exemplary Embodiment A-10. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-8, wherein the subject has received two previous anti-non-small cell lung cancer therapies. 38 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0261] Exemplary Embodiment A-11. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-10, wherein the subject has received at least one previous anti-non-small cell lung cancer therapy comprising at least one EGFR-targeting therapy.

[0262] Exemplary Embodiment A-12. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-11, wherein the subject has received at least one previous anti-non-small cell lung cancer therapy comprising at least one tyrosine kinase inhibitor (TKI).

[0263] Exemplary Embodiment A-13. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-12, wherein the subject has previously received osimertinib treatment.

[0264] Exemplary Embodiment A-14. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-13, wherein the subject has at least one non- classical EGFR mutation selected from EGFR-L833V, EGFR-R108K, EGFR-Y1016C, EGFR-L747_A750delinsP, EGFR-L747_P753delinsS, EGFR-L747_A755delinsSKD, EGFR-L747_T751del+V834L, EGFR-V834L, EGFR-K745N, EGFR- L747_P753delinsS+G930R, EGFR-G930R, EGFR-L747P_P753delinS, EGFR-L861Q, EGFR-L861R+L62R, EGFR-L62R, EGFR-L747_P753delinsS+L718V, EGFR- L747_P753delinsS+G930R, EGFR-L718V+G930R, EGFR-L747_T751del, EGFR-L861Q, EGFR-L861R, EGFR-E709_T710delInsD, EGFR-E709A, EGFR-L718V, EGFR-G719S, EGFR-G719A, EGFR-K745_E746insIPVAIK, EGFR-S768I, EGFR-V774M, EGFR- K745_E746insIPVAIK+K745N, EGFR-E19delinsD, and EGFR-Ex19insIPVAIK.

[0265] Exemplary Embodiment A-15. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-14, wherein the subject has at least one P-loop and α-C-helix compressing (PACC) mutation.

[0266] Exemplary Embodiment A-16. The method, compound for use, or use of Exemplary Embodiment 15, wherein the PACC mutation is selected from EGFR-E709_T710delInsD, EGFR-E709A, EGFR-L718V, EGFR-G719S, EGFR-G719A, EGFR- K745_E746insIPVAIK , EGFR-S768I, and EGFR-V774M.

[0267] Exemplary Embodiment A-17. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-16, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered daily. 39 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0268] Exemplary Embodiment A-18. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-17, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered once daily.

[0269] Exemplary Embodiment A-19. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-18, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered at a dosage of about 200 mg.

[0270] Exemplary Embodiment A-20. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-19, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is a mixture of Compound No.1A:(Compound No.1A), or a pharmaceutically acceptable salt thereof, and Compound No.1B:or a pharmaceutically acceptable salt thereof.

[0271] Exemplary Embodiment A-21. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-19, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1A: 40 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)(Compound No.1A), or a pharmaceutically acceptable salt thereof.

[0272] Exemplary Embodiment A-22. The method, compound for use, or use of Exemplary Embodiment 21, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1A.

[0273] Exemplary Embodiment A-23. The method, compound for use, or use of any one of Exemplary Embodiments A-1 through A-19, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1B:(Compound No.1B), or a pharmaceutically acceptable salt thereof.

[0274] Exemplary Embodiment A-24. The method, compound for use, or use of Exemplary Embodiment 23, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1B. EXAMPLES

[0275] It is understood that, unless indicated otherwise, the values presented in the examples are approximate values, and are subject to experimental and instrumental variations. Example 1. Exemplary Clinical Study of Compound No.1A.

[0276] A Phase 2 clinical study of Compound No.1A in patients with relapsed / refractory epidermal growth factor receptor (EGFR)-mutant (EGFRm) non-small cell lung cancer (NSCLC) was carried out, preliminary results of which are shown in FIGs.1-5. 41 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)

[0277] Patients across Cohorts 1 and 2 were administered with Compound No.1A. • Cohort 1: Patients with non-classical driver mutations after ≤2 prior lines of therapy with osimertinib as the preferred prior EGFR TKI; • Cohort 2: Patients with acquired resistance C797S after ≤2 prior lines of therapy with osimertinib as the only prior EGFR TKI.

[0278] To assess safety and tolerability, 40 patients were randomized to 100 mg (20 patients) or 200 mg (20 patients) daily dosage of Compound No.1A, across Cohorts 1 and 2. Compound No.1A showed a favorable tolerability profile (see FIG.2). The majority of adverse events were mild or moderate. For example, no grade 3 / 4 diarrhea, grade 4 rash, liver enzyme elevation, or QTc prolongation was observed.

[0279] Preliminary response rate and durability were assessed in 27 patients with 200 mg daily dosage of Compound No.1A for 21 days per cycle. Patient demographics and baseline characteristics are as shown in Table 4. Table 4. Patient Demographics and Baseline Characteristics (200 mg) *Onepatient had 3 prior lines of treatment (incl. amivantamab)

[0280] A broad spectrum of EGFR mutations were identified in patients who had received prior EGFR TKI therapy (see FIG.1).

[0281] Among the 27 patients, 22 response-evaluable patients met protocol eligibility criteria. For the 22 response-evaluable patients, the preliminary objective response rate (ORR) was 36%. Nineteen of these 22 patients expressed known osimertinib resistance mutations: either C797S or P-loop α-C helix compressing (PACC, a major subset of NCMs). Of these 19 patients, 8 achieved a response (42%): 5 with a confirmed partial response (cPR), including 1 patient who converted from a PR to an unconfirmed complete response (uCR) at 42 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) 8 months (and awaits confirmatory scan); and 3 with an unconfirmed partial response (uPR) at first scan and awaits a confirmatory scan. An additional 9 patients experienced stable disease (SD). See FIGs.3-4.

[0282] The percent change in sum of diameters of an omentum lesion in an exemplary patient is shown in FIG.5A. The patient has two non-classical mutations (G719S and S768I) and previously received 8 months of osimertinib treatment (first line) and 2 months of carboplatin and pemetrexed combination treatment (second line). The patient received over 8 months of treatment with 200 mg Compound No.1A administered once daily, and converted from a partial response to an unconfirmed complete response at 8 months and awaits confirmatory scan. FIGs.5C and 5D show scans of the patient’s omentum prior to Compound No.1A treatment (FIG.5C), and at Day 1 of Cycle 7 (FIG.5D). The lesion (or lack thereof) is indicated with an arrow, showing that the lesion is absent at Day 1 of Cycle 7 (FIG.5D). The presence of five other non-target lesions (pleura, LN-pleura, mesentery, and two brain lesions) were also monitored and observed to be absent by Month 8 of treatment (FIG.5B).

[0283] Further, Compound No.1A was seen to decrease EGFRm alleles and drive ctDNA clearance (see FIGs.6A-C). Eradication of targeted variant alleles and reduction of ctDNA are early predictors of progression-free survival (Thompson, JC., et al., British Journal of Cancer, 2023). FIGs.6A-6C shows the change in variant allele frequency percent (%VAF) and circulating tumor DNA percent (%ctDNA) clearance in three patients, comparing Day 1 of Cycle 1 (C1D1) and Day 1 of Cycle 3 (C3D1). All three patients had a confirmed partial response.

[0284] FIG.6A depicts a decrease in %VAF (C797S and E746_A750del) and %ctDNA clearance in a patient who previously received osimertinib treatment for 38.3 months.

[0285] FIG.6B depicts a decrease in %VAF (C797S, L833V, and L858R) and %ctDNA clearance in a patient who previously received osimertinib treatment for 50.0 months, followed by osimertinib, carboplatin, pemetrexed, and bevacizumab combination treatment for 26.6 months.

[0286] FIG.6C depicts a decrease in %VAF (C797, L858R, and Y1016C) and %ctDNA clearance in a patient who previously received carboplatin and paclitaxel combination treatment for 1.3 months, followed by osimertinib treatment for 16.8 months.

[0287] Durability was observed, with duration of response (DOR) of approximately 8 months or more for the first 3 patients with PR (see FIG.4).

[0288] Chemo-based combination regimens are administered via infusion, have high rates of grade 3 adverse effects (AEs), cover classical mutations, and are burdensome for the patients’ 43 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) quality of life. In contrast, Compound No.1A may be administered orally, be administered once daily, be well-tolerated, treat NSCLC with both classical and non-classical mutations, and may allow continued treatment of NSCLC via oral therapy post-osimertinib treatment. Thus, Compound No.1A may be a superior alternative to chemo-based combination regimens, such as those administered via infusion. EQUIVALENTS

[0289] The details of one or more embodiments of the disclosure are set forth in the accompanying description above. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Other features, objects, and advantages of the disclosure will be apparent from the description and from the claims. In the specification and the appended claims, the singular forms include plural referents unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications cited in this specification are incorporated by reference. The foregoing description has been presented only for the purposes of illustration and is not intended to limit the disclosure to the precise form disclosed, but by the claims appended hereto. 44 308418964

Claims

Attorney Docket No.43701-02302 (ASET-057 / 001WO) CLAIMS 1. A method of treating non-small cell lung cancer in a subject, comprising administering to the subject Compound No.1:(Compound No.1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

2. Compound No.1:(Compound No.1), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, for use in the treatment of non-small cell lung cancer in a subject.

3. Use of Compound No.1:a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the treatment of non- small cell lung cancer in a subject. 45 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) 4. Use of Compound No.1:a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for the treatment of non-small cell lung cancer in a subject.

5. The method, compound for use, or use of any one of claims 1-4, wherein the non-small cell lung cancer is metastatic non-small cell lung cancer .

6. The method, compound for use, or use of claim 5, wherein the non-small cell lung cancer has metastasized to the brain.

7. The method, compound for use, or use of any one of claims 1-4, wherein the non-small cell lung cancer is locally advanced non-small cell lung cancer.

8. The method, compound for use, or use of any one of claims 1-7, wherein the subject has previously received at least one anti-non-small cell lung cancer therapy, wherein the subject has: i) at least one classical EGFR mutation and a EGFR-C797S mutation; ii) at least one classical EGFR mutation and at least one non-classical EGFR mutation; iii) at least one classical EGFR mutation, at least one non-classical EGFR mutation, and a EGFR-C797S mutation; or iv) a EGFR-C797S mutation.

9. The method, compound for use, or use of any one of claims 1-8, wherein the subject has received one previous anti-non-small cell lung cancer therapy. 46 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) 10. The method, compound for use, or use of any one of claims 1-8, wherein the subject has received two previous anti-non-small cell lung cancer therapies.

11. The method, compound for use, or use of any one of claims 1-10, wherein the subject has received at least one previous anti-non-small cell lung cancer therapy comprising at least one EGFR-targeting therapy.

12. The method, compound for use, or use of any one of claims 1-11, wherein the subject has received at least one previous anti-non-small cell lung cancer therapy comprising at least one tyrosine kinase inhibitor (TKI).

13. The method, compound for use, or use of any one of claims 1-12, wherein the subject has previously received osimertinib treatment.

14. The method, compound for use, or use of any one of the preceding claims, wherein the subject has at least one non-classical EGFR mutation selected from EGFR-L833V, EGFR- R108K, EGFR-Y1016C, EGFR-L747_A750delinsP, EGFR-L747_P753delinsS, EGFR- L747_A755delinsSKD, EGFR-L747_T751del+V834L, EGFR-V834L, EGFR-K745N, EGFR-L747_P753delinsS+G930R, EGFR-G930R, EGFR-L747P_P753delinS, EGFR-L861Q, EGFR-L861R+L62R, EGFR-L62R, EGFR-L747_P753delinsS+L718V, EGFR- L747_P753delinsS+G930R, EGFR-L718V+G930R, EGFR-L747_T751del, EGFR-L861Q, EGFR-L861R, EGFR-E709_T710delInsD, EGFR-E709A, EGFR-L718V, EGFR-G719S, EGFR-G719A, EGFR-K745_E746insIPVAIK, EGFR-S768I, EGFR-V774M, EGFR- K745_E746insIPVAIK+K745N, EGFR-E19delinsD, and EGFR-Ex19insIPVAIK.

15. The method, compound for use, or use of any one of the preceding claims, wherein the subject has at least one P-loop and α-C-helix compressing (PACC) mutation.

16. The method, compound for use, or use of claim 15, wherein the PACC mutation is selected from EGFR-E709_T710delInsD, EGFR-E709A, EGFR-L718V, EGFR-G719S, EGFR-G719A, EGFR-K745_E746insIPVAIK , EGFR-S768I, and EGFR-V774M.

17. The method, compound for use, or use of any one of the preceding claims, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered daily. 47 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO) 18. The method, compound for use, or use of any one of the preceding claims, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered once daily.

19. The method, compound for use, or use of any one of the preceding claims, wherein Compound No.1, or the pharmaceutically acceptable salt thereof, is administered at a dosage of about 200 mg.

20. The method, compound for use, or use of any one of the preceding claims, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is a mixture of Compound No.1A:(Compound No.1A), or a pharmaceutically acceptable salt thereof, and Compound No.1B:(Compound No.1B), or a pharmaceutically acceptable salt thereof.

21. The method, compound for use, or use of any one of claims 1-19, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1A: 48 308418964Attorney Docket No.43701-02302 (ASET-057 / 001WO)(Compound No.1A), or a pharmaceutically acceptable salt thereof.

22. The method, compound for use, or use of claim 21, wherein Compound No. 1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1A.

23. The method, compound for use, or use of any one of claims 1-19, wherein Compound No.1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1B:(Compound No.1B), or a pharmaceutically acceptable salt thereof.

24. The method, compound for use, or use of claim 23, wherein Compound No. 1, the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, is Compound No.1B. 49 308418964

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