Mutation selective EGFR inhibitors

Abstract

The present disclosure relates to compounds that are epidermal growth factor receptor (EGFR) allosteric inhibitors; pharmaceutical compositions comprising such compounds; and methods of treating or preventing kinase-mediated conditions, including cancer and other proliferative diseases.

Description

[0001] Related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 479,487, filed on January 11, 2023, the entire contents of which are hereby incorporated by reference.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under Grant Nos. R01CA201049 and F32CA247198 awarded by the National Institutes of Health. The government has certain rights in this invention. Background Art

[0003] Epidermal growth factor receptor (EGFR, Erb-B1) belongs to the receptor tyrosine kinase family, which mediates the proliferation, differentiation and survival of normal and malignant cells (Arteaga, CL, J. Clin. Oncol. 19, 2001, 32-40). EGFR dysregulation is associated with many types of human cancers, with overexpression of this receptor occurring in at least 70% of human cancers (Seymour, LK, Curr. Drug Targets 2, 2001, 117-133), including non-small cell lung cancer, breast cancer, glioma, head and neck squamous cell carcinoma and prostate cancer (Raymond, E., et al., Drugs 60 (Suppl 1), 2000, 15-23, discussion 41-2; Salomon, DS, et al., Crit. Rev. Oncol. Hematol . 19, 1995, 183-232; Voldborg B. R., et al., Ann. Oncol. 8, 1997, 1197-1206). Therefore, EGFR has become an attractive target for the design and development of diagnostic and therapeutic drugs that can specifically bind to and inhibit the receptor's tyrosine kinase activity and signal transduction pathways in cancer cells. The two most common activating EGFR mutations in patients with non-small cell lung cancer (NSCLC) are exon 19 deletion (del19) and the L858R point mutation.

[0004] The reversible EGFR inhibitors gefitinib and erlotinib are effective clinical treatments for patients with advanced non-small cell lung cancer (NSCLC) harboring EGFR mutations (Mok, TS, et al., N. Engl. J. Med. 361, 2009, 947-57; Paez, JG, et al., Science 304, 2004, 1497-500; Lynch, TJ, et al., N. Engl. J. Med. 350,2004, 2129-39;Rosell, R.,et al., Lancet Oncol. 13, 2012, 239-46). However, most patients experience disease progression after treatment with these drugs. In 60% of patients, the most common mechanism of acquired resistance is a secondary mutation at position T790 of EGFR (T790M) (Yu, HA, et al., Clin. Cancer Res. 19, 2013, 2240-7). This mutation results in an increased affinity for ATP, making it more difficult for the reversible EGFR tyrosine kinase inhibitors (TKIs) gefitinib and erlotinib to bind to the EGFR TKI domain (Yun CH, et al., Proc. Natl. Acad. Sci. USA 105, 2008,2070-5).

[0005] Covalent EGFR inhibitors were developed to address resistance caused by the EGFR T790M mutation. One such covalent EGFR inhibitor is afatinib, which is active against both mutant and wild-type (WT) EGFR (Li, D.; et al., Oncogene 27, 2008 (4702-2711). However, inhibition of WT EGFR can lead to toxicities including rash and diarrhea, which may be dose-limiting in the clinic. (Yap, TA; et al., J Clin Onc , 28, 2010 (3965-3972).

[0006] The third generation of covalent EGFR inhibitors that are selective for mutant EGFR but not WT EGFR have overcome the limitations of afatinib and have shown better clinical efficacy and tolerability (Cross, DEA; et al., Cancer Discussion . 4, 2014 (1046-1061). Osimertinib was initially approved for the treatment of patients with T790M mutation-resistant reversible inhibitors and is currently approved for first-line treatment. Although the main resistance mechanism of first-generation inhibitors has been addressed, resistance to osimertinib has also emerged (Schmid, S., et al., Lung Cancer 147, 2020, 123-129). Resistance mechanisms include the C797S mutation, which prevents osimertinib from forming a covalent bond with a cysteine ​​residue (Thress, KT, et al., Nature Med This resistance can lead to triple mutations (del19 / T790M / C797S or L858R / T790M / C797S) or double mutations (e.g., L858R / C797S) in first-line therapy, which are unsuccessful with currently approved EGFR inhibitors.

[0007] Therefore, new targeted therapies are still needed for NSCLC patients who have developed resistance to existing inhibitors. Summary of the Invention

[0008] In one aspect, provided herein are compounds of Formula I: or a pharmaceutically acceptable salt thereof.

[0009] In another aspect, provided herein is a compound of Formula II: or a pharmaceutically acceptable salt thereof.

[0010] In another aspect, provided herein is a pharmaceutical composition comprising a compound disclosed herein or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

[0011] In yet another aspect, provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein.

[0012] In yet another aspect, provided herein is a method of inhibiting a kinase in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein.

[0013] In one aspect, provided herein is a method of treating a kinase-mediated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein. DETAILED DESCRIPTION

[0014] definition Listed below are definitions of various terms used to describe the compounds and compositions disclosed herein. These definitions apply to the terms as used in this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0015] Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well known and commonly used in the art.

[0016] As used herein, the articles "a" and "an" refer to one or more than one (i.e., at least one) of the grammatical object of the article. For example, "an element" means one element or more than one element. In addition, the use of the term "including" and other forms, such as "includes" and "included" is not limited.

[0017] As used herein, the term "about" will be understood by one of ordinary skill in the art and will vary depending on the context in which it is used. As used herein, when referring to a measurable value such as an amount, a duration of time, and the like, the term "about" is intended to encompass variations of ±20% or ±10%, including ±5%, ±1%, and ±0.1%, from the specified value, as such variations are suitable for performing the disclosed methods.

[0018] As used herein, the term "administering" or similar terms refers to providing a therapeutic agent to a subject. There are a variety of techniques for administering therapeutic agents in the art, including but not limited to intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0019] The terms "treat / treated / treating / treatment" include reducing or alleviating at least one symptom associated with or caused by the state, disorder or disease being treated. In certain embodiments, treatment comprises contacting an effective amount of a compound disclosed herein with wild-type or mutant EGFR for treating a condition associated with cancer.

[0020] As used herein, the terms "prevent" and "prevention" mean that a condition or disease will not develop if it has not already occurred, or will not develop further if it has already developed. The ability to prevent some or all of the symptoms associated with a condition or disease is also contemplated.

[0021] As used herein, the terms "patient," "individual," or "subject" refer to a human or non-human mammal. Non-human mammals include, for example, livestock and pets such as sheep, bovines, swine, canines, felines, and marine mammals. Preferably, the patient, subject, or individual is a human.

[0022] As used herein, the terms "effective amount," "pharmaceutically effective amount," and "therapeutically effective amount" refer to a nontoxic but sufficient amount of an agent to provide a desired biological result. The result can be a reduction or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. The appropriate therapeutic amount in any individual case can be determined by one of ordinary skill in the art using routine experimentation.

[0023] As used herein, the term "pharmaceutically acceptable" refers to a material, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compound and is relatively nontoxic, i.e., the material can be administered to an individual without causing adverse biological effects or interacting in a deleterious manner with any component of the composition in which it is contained.

[0024] As used herein, the term "pharmaceutically acceptable salt" refers to a derivative of a disclosed compound in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues (such as amines); alkali metal or organic salts of acidic residues (such as carboxylic acids); and the like. Pharmaceutically acceptable salts of the present disclosure include, for example, conventional non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of the present disclosure can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base form of these compounds with an appropriate base or acid in water or in an organic solvent or in a mixture of the two; typically, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. The phrase "pharmaceutically acceptable salt" is not limited to monosalts or 1:1 salts. For example, "pharmaceutically acceptable salt" also includes disalts, such as dihydrochloride. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is herein incorporated by reference in its entirety.

[0025] As used herein, the term "prodrug" refers to a precursor compound that will undergo metabolic activation in vivo to produce an active drug. Thus, for example, a prodrug of a compound provided herein will undergo metabolic activation to produce the compound when administered to a subject.

[0026] As used herein, the term "composition" or "pharmaceutical composition" refers to a mixture of at least one compound useful in the present disclosure and a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. A variety of techniques exist in the art for administering compounds, including but not limited to intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0027] As used herein, the term "pharmaceutical combination" means a product resulting from the mixing or combining of more than one active ingredient, and includes fixed and non-fixed combinations of the active ingredients. The term "fixed combination" means that both the active ingredients, such as a compound of the present disclosure and a co-agent, are administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that both the active ingredients, such as a compound of the present disclosure and a co-agent, are administered to a patient as separate entities simultaneously, concurrently or sequentially, but without specific time limits, wherein such administration provides therapeutically effective levels in the patient. The latter also applies to cocktail therapy, for example, the administration of three or more active ingredients.

[0028] As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, that is involved in carrying or transporting a compound useful in the present disclosure in or to a patient, enabling it to perform its intended function. Typically, such constructs are carried or transported from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation (including the compounds useful in the present disclosure) and not causing harm to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffered saline; and other nontoxic, compatible substances used in pharmaceutical formulations.

[0029] As used herein, "pharmaceutically acceptable carrier" also includes any and all coating agents, antibacterial and antifungal agents, and absorption delaying agents, etc., which are compatible with the activity of the compounds useful in the present disclosure and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the composition. "Pharmaceutically acceptable carrier" may also include pharmaceutically acceptable salts of the compounds disclosed herein. Other additional ingredients that may be included in pharmaceutical compositions are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

[0030] As used herein, the term "EGFR" refers to epidermal growth factor receptor (also known as ErbB-1 or HER1) and can refer to a wild-type receptor or a receptor containing one or more mutations.

[0031] As used herein, the term "HER" or "Her" refers to members of the ErbB receptor tyrosine kinase family, including EGFR, ERBB2, HER3, and HER4.

[0032] As used herein, the term "allosteric site" refers to a site on EGFR other than the ATP binding site, such as the site characterized in the EGFR crystal structure. An "allosteric site" can be a site near the ATP binding site, such as the site characterized in the EGFR crystal structure. For example, an allosteric site includes one or more of the following amino acid residues of epidermal growth factor receptor (EGFR): Lys745, Leu788, Ala743, Cys755, Leu777, Phe856, Asp855, Met766, Ile759, Glu762 and / or Ala763.

[0033] As used herein, the term "agent that prevents EGFR dimer formation" or its iterations refers to an agent that prevents dimer formation in which the C lobe of the "activator" subunit collides with the N lobe of the "receptor" subunit. Examples of agents that prevent EGFR dimer formation include, but are not limited to, cetuximab, trastuzumab, panitumumab, and Mig6.

[0034] As used herein, the term "alkyl", by itself or as part of another substituent, unless otherwise indicated, means a straight or branched saturated hydrocarbon radical having the specified number of carbon atoms (i.e., C1-C6 alkyl means an alkyl radical having from one to six carbon atoms), and includes straight and branched chains. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, and hexyl. Other examples of C1-C6 alkyl include ethyl, methyl, isopropyl, isobutyl, n-pentyl, and n-hexyl.

[0035] As used herein, the term "haloalkyl" refers to an alkyl group as defined above, substituted with one or more halo substituents, wherein alkyl and halo are as defined herein. Haloalkyl includes, for example, chloromethyl, trifluoromethyl, bromoethyl, chlorofluoroethyl, and the like.

[0036] As used herein, the term "alkoxy" refers to the group -O-alkyl, wherein alkyl is as defined above. Alkoxy includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, and the like.

[0037] As used herein, the term "alkylamine" refers to the group -NH-alkyl, wherein alkyl is as defined above. Alkylamines include, for example, methylamine, ethylamine, isopropylamine, n-propylamine, n-butylamine, sec-butylamine, tert-butylamine, and the like.

[0038] As used herein, the term "haloalkoxy" refers to the group -O-haloalkyl, wherein haloalkyl is as defined above. Haloalkoxy includes, for example, chloromethoxy, trifluoromethoxy, bromoethoxy, chlorofluoroethoxy, and the like.

[0039] As used herein, the term "alkenyl" refers to a monovalent group derived from a hydrocarbon moiety, which, in certain embodiments, contains 2 to 6, or 2 to 8 carbon atoms and has at least one carbon-carbon double bond. An alkenyl group may or may not be the point of attachment to another group. The term "alkenyl" includes, but is not limited to, vinyl, 1-propenyl, 1-butenyl, heptenyl, octenyl, etc.

[0040] As used herein, the term "alkynyl" refers to a monovalent group derived from a hydrocarbon moiety, which, in certain embodiments, contains 2 to 6, or 2 to 8, carbon atoms and has at least one carbon-carbon triple bond. An alkynyl group may or may not be the point of attachment to another group. The term "alkynyl" includes, but is not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

[0041] As used herein, the term "halo" or "halogen," by itself or as part of another substituent, means, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom, preferably fluorine, chlorine, or bromine, more preferably fluorine or chlorine.

[0042] As used herein, the term "cycloalkyl" means a fully saturated non-aromatic carbocyclic ring system with 1, 2 or 3 rings, wherein such rings can be fused. The term "fused" means a second ring that exists (i.e., is connected or formed) by having two adjacent atoms that are shared (i.e., shared) with the first ring. Cycloalkyl also includes a bicyclic structure, the properties of which can be bridged or spirocyclic, wherein each individual ring in the bicyclic has 3 to 8 atoms. The term "cycloalkyl" includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [3.1.0] hexyl, spiro [3.3] heptyl and bicyclo [1.1.1] pentyl. In one embodiment, cycloalkyl is a 3-10 yuan cycloalkyl. In another embodiment, cycloalkyl is a 3-6 yuan cycloalkyl.

[0043] As used herein, the term "cycloalkenyl" refers to a partially saturated non-aromatic carbocyclic ring system having 1, 2 or 3 rings, wherein such rings may be fused, and wherein at least one ring contains sp 2 Carbon-carbon bond. The term "cycloalkenyl" includes, but is not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, bicyclo[3.1.0]hexenyl, spiro[3.3]heptenyl, and bicyclo[1.1.1]pentenyl. In one embodiment, a cycloalkenyl is a 3-10 membered cycloalkyl. In another embodiment, a cycloalkenyl is a 4-7 membered cycloalkyl.

[0044] As used herein, the term "heterocyclyl" or "heterocycloalkyl" means a non-aromatic carbocyclic ring system containing 1, 2, 3 or 4 heteroatoms independently selected from N, O and S and having 1, 2 or 3 rings, wherein the rings may be fused, wherein fusion is as defined above. Heterocyclyl also includes bicyclic structures, which may be bridged or spirocyclic in nature, wherein each individual ring within the bicyclic ring has 3 to 8 atoms and contains 0, 1 or 2 N, O or S atoms. The term "heterocyclyl" includes cyclic esters (i.e., lactones) and cyclic amides (i.e., lactams), and also specifically includes, but is not limited to, epoxide, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl (i.e., oxanyl), pyranyl, dioxanyl, aziridinyl, pyrrolidinyl, 2,5-dihydro-1H-pyrrolyl, oxazolidinyl, thiazolidinyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, 1,3-oxazinyl, 1,3-thiazinyl, 2-azabicyclo[2.1.1]hexyl, 5-azabicyclo[2.1.1]hexyl, 6-azabicyclo[3.1.1]heptyl, 2-azabicyclo[2.2.1]heptyl, 3-azabicyclo[3.1.1]heptyl, 2-azabicyclo[3.1 .1]heptyl, 3-azabicyclo[3.1.0]hexyl, 2-azabicyclo-[3.1.0]hexyl, 3-azabicyclo[3.2.1]octyl, 8-azabicyclo[3.2.1]octyl, 3-oxa-7-azabicyclo[3.3.1]-nonyl, 3-oxa-9-azabicyclo[3.3.1]nonyl, 2-oxa-5-azabicyclo[2.2.1]heptyl, 6-oxa-3-aza-bicyclo[3.1.1]heptyl, 2-azaspiro[3.3]heptyl, 2-oxa-6-azaspiro[3.3]heptyl, 2-oxaspiro-[3.3]heptyl, 2-oxaspiro[3.5]nonyl, 3-oxaspiro[5.3]nonyl and 8-oxabicyclo[3.2.1]octyl. In one embodiment, heterocycloalkyl is a 3-10 membered heterocycloalkyl.In another embodiment, heterocycloalkyl is a 4-7 membered heterocyclyl.

[0045] As used herein, the term "aromatic" refers to a carbocyclic or heterocyclic ring having one or more polyunsaturated rings and having aromatic character (ie, having (4n+2) delocalized π (pi) electrons, where n is an integer).

[0046] As used herein, the term "aryl" means an aromatic carbocyclic ring system containing 1, 2, or 3 rings, wherein such rings may be fused, wherein fusion is as defined above. If the rings are fused, one of the rings must be completely unsaturated, and one or more fused rings may be completely saturated, partially unsaturated, or completely unsaturated. The term "aryl" includes, but is not limited to, phenyl, naphthyl, indanyl, and 1,2,3,4-tetrahydronaphthyl. In some embodiments, the aryl group has 6 carbon atoms. In some embodiments, the aryl group has six to ten carbon atoms. In some embodiments, the aryl group has six to sixteen carbon atoms.

[0047] As used herein, the term "heteroaryl" means an aromatic carbocyclic ring system containing 1, 2, 3 or 4 heteroatoms independently selected from N, O and S and having 1, 2 or 3 rings, wherein such rings may be fused, wherein fusion is as defined above. The term "heteroaryl" includes, but is not limited to, furanyl, thienyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, imidazo[1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, 5,6,7,8-tetrahydroisoquinolyl, 5,6,7,8-tetrahydroquinolyl, 6,7-dihydro-5H-cyclopenta[b]pyridinyl, 6,7-dihydro-5H-cyclopenta[c]pyridinyl, 1,4, 5,6-tetrahydrocyclopenta[c]pyrazolyl, 2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl, 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazolyl, 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]-triazolyl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridinyl, 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridinyl, 4,5,6,7-tetrahydro-1H-indazolyl and 4,5,6,7-tetrahydro-2H-indazolyl. In one embodiment, the heteroaryl group is a 5-10 membered heteroaryl group. In another embodiment, the heteroaryl group is a 5-6 membered heteroaryl group.

[0048] It should be understood that if an aryl, heteroaryl, cycloalkyl, or heterocyclyl moiety can be bonded or otherwise attached to a designated moiety through different ring atoms (i.e., shown or described without indicating a specific point of attachment), all possible points are intended to be represented, whether through a carbon atom or, for example, a trivalent nitrogen atom. For example, the term "pyridyl" means 2-, 3-, or 4-pyridyl, the term "thienyl" means 2- or 3-thienyl, and so on.

[0049] As used herein, the term "substituted" means that an atom or group of atoms has replaced a hydrogen as a substituent attached to another group.

[0050] As used herein, the term "optionally substituted" means that the referenced group may be substituted or unsubstituted. In one embodiment, the referenced group is optionally substituted with zero substituents, i.e., the referenced group is unsubstituted. In another embodiment, the referenced group is optionally substituted with one or more additional groups individually and independently selected from the groups described herein.

[0051] Compound Provided herein are compounds that are allosteric inhibitors of epidermal growth factor receptor (EGFR) that are useful in treating kinase-mediated disorders, including cancer and other proliferative diseases.

[0052] In one aspect, provided herein are compounds of Formula I: or a pharmaceutically acceptable salt thereof; in: represents a single bond or a double bond; A and A' are each independently CH, CR 10 , CH2, O or N; W and Z are each independently N or CR 9 ; X and Y are each independently N, CH or CR 3 ; provided that at least one of W, X, Y or Z is CH; R 1 is selected from the group consisting of 6-10 membered aryl, 5-10 membered heteroaryl, 3-10 membered heterocycloalkyl and 3-10 membered cycloalkyl, all of which are optionally substituted by one, two or three R 8 replace; Each R 2 independently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0- a group consisting of 2NH2, CN, and a 3-10 membered cycloalkyl group; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl group or a 3-10 membered heterocycloalkyl group; R 3 is independently selected at each occurrence from halogen, OR 4 NR 4 R 4 、SO2R4 、SO2NHR 4 NHSO2R 4 、C(O)OR 4 、C(O)NHR 4 、C(O)R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, C4-C7 cycloalkenyl, C6-C 10 The group consisting of aryl, 5-6 membered heteroaryl and 4-7 membered heterocyclic group, wherein alkyl, alkenyl or alkynyl are each optionally replaced by R 4 substituted once, twice or three times, and wherein aryl, heteroaryl or heterocyclyl are each optionally replaced by R 5 Replace once, twice or thrice; R 4 is independently selected at each occurrence from H, C1-C6 alkyl, (CH2) 0-3 -(C3-C7 cycloalkyl), (CH2) 0-3 -(C4-C7 cycloalkenyl), (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 6 Replace once, twice or thrice; R 5 is independently selected at each occurrence from C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C3 alkylamine, 3-10 membered cycloalkyl, halogen, COOH, C(O)O(C1-C6 alkyl), O(CH2) 1-3 -OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, OH, CN, (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 7 Replace once, twice or thrice; R 6 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4OH, S(O) 0- 2H、S(O) 0-2 A group consisting of NH2 or CN; R 7 is independently selected at each occurrence from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, halogen, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, SO2NH2, SO2NH(C1-C6 alkyl), SO2N(C1-C6 alkyl)2, (CH2) 1-2 -OH, C(O)(CH2) 1-2 -OH, C(O)(C1-C6 alkyl) and C(O)O(C1-C6 alkyl) substituents; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 The group composed of NH2 and CN; R 9 is independently selected at each occurrence from the group consisting of H, halo, C1-C3 alkyl, C1-C3 haloalkyl, and C1-C3 alkoxy; R 10 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 the group consisting of NH2 and CN; and n is 0, 1, 2 or 3.

[0053] In yet another embodiment, In another embodiment, Indicates a double bond.

[0054] In one embodiment, A is CH2. In another embodiment, A is CH. In yet another embodiment, at least one of A and A' is CH or CR 10 In yet another embodiment, at least one of A and A' is CH, CR 10or CH2. In one embodiment, A' is N. In another embodiment, A' is CH. In yet another embodiment, A' is CH2.

[0055] In one embodiment, W is N. In another embodiment, W is CR 9 In one embodiment, W is CH or C-halo. In yet another embodiment, Z is N. In yet another embodiment, Z is CR 9 In one embodiment, Z is CH or C-halo.

[0056] In another embodiment, when X is CR 3 In yet another embodiment, when Y is CR 3 In another embodiment, X is CR 3 In one embodiment, Y is CR 3 .

[0057] In one embodiment, R 1 is optionally replaced by one, two or three R 8 In another embodiment, R 1 is optionally replaced by one, two or three R 8 In another embodiment, R 1 is thiazolyl or pyridinyl, both optionally substituted by one, two or three R 8 In yet another embodiment, R 1 Selected from the group consisting of: or ; Both are optionally replaced by one, two or three R 8 replace.

[0058] In yet another embodiment, the compound of formula I is a compound of formula Ia: or a pharmaceutically acceptable salt thereof.

[0059] In one embodiment, the compound of formula (I) is a compound of formula Ib: or a pharmaceutically acceptable salt thereof.

[0060] In another embodiment, the compound of formula I is a compound of formula Id: or a pharmaceutically acceptable salt thereof.

[0061] In another embodiment, R 2 is halogen or C1-C3 alkyl. In yet another embodiment, R 2 In another embodiment, R 2 is C1-C3 alkyl. In another embodiment, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl or a 3-10 membered heterocycloalkyl. 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl group.

[0062] In one embodiment, R 3 Is one or two R 5 In another embodiment, R 3 Is one or two R 5 In yet another embodiment, R 5 is optionally replaced by R 7 substituted once, twice or three times 4-7 membered heterocyclyl. In one embodiment, R 5 is optionally replaced by R 7 In another embodiment, R 5 is piperidinyl or piperazinyl, both of which are optionally replaced by R 7 In one embodiment, R 3 yes or .

[0063] In another embodiment, R 7 In yet another embodiment, R 7 It's methyl.

[0064] In yet another embodiment, R 4 is independently at each occurrence selected from the group consisting of H and C1-C6 alkyl.

[0065] In yet another embodiment, R 9 is H, halo or C1-C3 haloalkyl. In one embodiment, R 9 is H. In another embodiment, R 9 In yet another embodiment, R 9 In another embodiment, R 9 It is a C1-C3 alkoxy group.

[0066] In yet another embodiment, n is 0 or 1. In one embodiment, n is 0. In another embodiment, n is 1.

[0067] In yet another embodiment, represents a single bond or a double bond; A and A' are each independently CH, CR2, O or N; W and Z are each independently CH or CR 9 ; X and Y are each independently CH or CR 3 ; R 1 is optionally replaced by one, two or three R 8 substituted 5-10 membered heteroaryl; R 2 is halogen or C1-C3 alkyl; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl group or a 3-10 membered heterocycloalkyl group; R 3 Is one or two R 5 substituted 6-10 membered aryl; R 5 is optionally replaced by R 7 a 4-7 membered heterocyclic group substituted once, twice or three times; R 7 is a C1-C6 alkyl group; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 The group composed of NH2 and CN; R 9 is independently selected at each occurrence from the group consisting of halo, C1-C3 alkyl, C1-C3 haloalkyl, and C1-C3 alkoxy; and n is 0 or 1.

[0068] In another aspect, provided herein is a compound of Formula II: or a pharmaceutically acceptable salt thereof; A and A' are each independently CH, CR 10 or N; R 1 is selected from the group consisting of 6-10 membered aryl, 5-10 membered heteroaryl, 3-10 membered heterocycloalkyl and 3-10 membered cycloalkyl, all of which are optionally substituted by one, two or three R 8 replace; Each R 2 independently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0- a group consisting of 2NH2, CN, and a 3-10 membered cycloalkyl group; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl group or a 3-10 membered heterocycloalkyl group; R 3 is independently selected at each occurrence from halogen, OR 4 NR 4 R 4 、SO2R 4 、SO2NHR 4 NHSO2R 4 、C(O)OR 4 、C(O)NHR 4 、C(O)R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, C4-C7 cycloalkenyl, C6-C 10 The group consisting of aryl, 5-6 membered heteroaryl and 4-7 membered heterocyclic group, wherein alkyl, alkenyl or alkynyl are each optionally replaced by R 4 substituted once, twice or three times, and wherein aryl, heteroaryl or heterocyclyl are each optionally replaced by R 5 Replace once, twice or thrice; R 4 is independently selected at each occurrence from H, C1-C6 alkyl, (CH2) 0-3 -(C3-C7 cycloalkyl), (CH2) 0-3 -(C4-C7 cycloalkenyl), (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 6 Replace once, twice or thrice; R 5 is independently selected at each occurrence from C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C3 alkylamine, 3-10 membered cycloalkyl, halogen, COOH, C(O)O(C1-C6 alkyl), O(CH2) 1-3 -OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, OH, CN, (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 7 Replace once, twice or thrice; R 6 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0- 2H、S(O) 0-2 A group consisting of NH2 or CN; R 7 is independently selected at each occurrence from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, halogen, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, SO2NH2, SO2NH(C1-C6 alkyl), SO2N(C1-C6 alkyl)2, (CH2) 1-2 -OH, C(O)(CH2) 1-2 -OH, C(O)(C1-C6 alkyl) and C(O)O(C1-C6 alkyl) substituents; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 The group composed of NH2 and CN; R 10is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 the group consisting of NH2 and CN; and n is 0, 1, 2 or 3.

[0069] In one embodiment, A is N. In another embodiment, A is N. In yet another embodiment, A is CH 8 In one embodiment, A' is N. In yet another embodiment, A' is CH. In yet another embodiment, at least one of A and A' is CH or CR 8 .

[0070] In one embodiment, R 1 is optionally replaced by one, two or three R 8 In another embodiment, R 1 is optionally replaced by one, two or three R 8 In yet another embodiment, R 1 is thiazolyl or pyridinyl, both optionally substituted by one, two or three R 8 In another embodiment, R 1 Selected from the group consisting of: and ; Both are optionally replaced by one, two or three R 8 replace.

[0071] In one embodiment, the compound of formula II is a compound of formula IIa: or a pharmaceutically acceptable salt thereof.

[0072] In one embodiment, R 2 In another embodiment, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl or a 3-10 membered heterocycloalkyl. 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl group.

[0073] In another embodiment, R 3 Is one or two R5 In yet another embodiment, R 3 Is one or two R 5 In another embodiment, R 5 is optionally replaced by R 7 substituted once, twice or three times 4-7 membered heterocyclyl. In one embodiment, R 5 is optionally replaced by R 7 In another embodiment, R 3 yes, or .

[0074] In yet another embodiment, R 7 In another embodiment, R 7 It's methyl.

[0075] In one embodiment, A and A' are each independently CH or N; R 1 is optionally replaced by one, two or three R 8 substituted 5-10 membered heteroaryl; R 2 It is a halogen; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl group or a 3-10 membered heterocycloalkyl group; R 3 Is one or two R 5 substituted 6-10 membered aryl; R 5 is optionally replaced by R 7 a 4-7 membered heterocyclic group substituted once, twice or three times; R 7 is a C1-C6 alkyl group; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 the group consisting of NH2 and CN; and n is 0 or 1.

[0076] In another embodiment, the compound of Formula I or II is selected from the group consisting of the compounds in Table 1.

[0077] Table 1. or a pharmaceutically acceptable salt thereof.

[0078] The compounds disclosed herein may exist as tautomers and optical isomers (eg, enantiomers, diastereomers, mixtures of diastereomers, racemic mixtures, etc.).

[0079] Compounds provided herein may also include all isotopes of atoms present in intermediates or final compounds. Isotopes include atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be replaced or substituted by isotopes of atoms of natural or non-natural abundance. In some embodiments, the compound comprises at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced or substituted by deuterium. In some embodiments, the compound comprises two or more deuterium atoms. In some embodiments, the compound comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 deuterium atoms. Methods for incorporating isotopes into the synthesis of organic compounds are known in the art (Deuterium Labeling in Organic Chemistry, ed. Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971); The Renaissance of H / D Exchange, ed. Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling, ed. James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds are useful in various studies, such as NMR spectroscopy, metabolic experiments and / or analysis.

[0080] In the compounds provided herein, any atom that is not explicitly designated as a specific isotope is intended to represent any stable isotope of the atom. Unless otherwise stated, when a position is specifically designated as "H" or "hydrogen", the position is understood to have hydrogen at its natural abundance isotopic composition. In addition, unless otherwise stated, when a position is specifically designated as "D" or "deuterium", it is understood that the deuterium abundance at the position is at least 3000 times the natural abundance of deuterium (which is 0.015%) (i.e., at least 45% deuterium incorporation).

[0081] It is generally known in the art that any compound that will be converted in vivo to provide a compound disclosed herein is a prodrug within the scope of this disclosure.

[0082] In one aspect, provided herein is a pharmaceutical composition comprising any one of the compounds disclosed herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0083] In one embodiment, the composition further comprises a second active agent. In another embodiment, the second active agent is selected from the group consisting of a MEK inhibitor, a PI3K inhibitor, and an mTor inhibitor. In yet another embodiment, the second active agent can prevent the formation of EGFR dimers in the subject. In yet another embodiment, the second active agent is selected from the group consisting of cetuximab, trastuzumab, and panitumumab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib, or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0084] In another aspect, provided herein is a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In another aspect, the pharmaceutical composition further comprises a second active agent, wherein the second active agent prevents EGFR dimer formation, and a pharmaceutically acceptable carrier. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab.

[0085] Compounds that are bound to the allosteric site in EGFR, such as compounds of the present disclosure (e.g., compounds of the formula disclosed herein), are optionally combined with a second active agent, wherein the second active agent prevents EGFR dimer formation and can regulate EGFR activity. In some embodiments, compounds of the present disclosure can inhibit or reduce EGFR activity without the need for a second active agent (e.g., an antibody such as cetuximab, trastuzumab, or panitumumab). In other embodiments, compounds of the present disclosure are combined with a second active agent. In one embodiment, the second active agent prevents EGFR dimer formation and / or can inhibit or reduce EGFR activity. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib, or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0086] Treatment In one aspect, the present invention provides a method of treating a cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein. In one embodiment, the cancer is selected from the group consisting of lung cancer, colon cancer, breast cancer, endometrial cancer, thyroid cancer, glioma, squamous cell carcinoma, and prostate cancer. In another embodiment, the cancer is non-small cell lung cancer (NSCLC).

[0087] In another aspect, provided herein is a method of inhibiting a kinase in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound provided herein. In one embodiment, the kinase is EGFR.

[0088] In yet another aspect, provided herein is a method for treating or preventing a kinase-mediated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure. In one embodiment, the kinase-mediated disorder is resistant to an EGFR-targeted therapy. In another embodiment, the EGFR therapy is selected from the group consisting of gefitinib, erlotinib, or osimertinib.

[0089] In some embodiments, the compound of the present disclosure can regulate (for example, suppress or reduce) the activity of the EGFR containing one or more mutations. In some embodiments, the mutant EGFR contains one or more mutations selected from T790M, L718Q, L844V, V948R, L858R, I941R and C797S. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from L858R / L718Q, L858R / L844V, L858R / T790M, L858R / T790M / I941R, L858R / C797S, L858R / T790M / C797S and L858R / T790M / L718Q. In other embodiments, the mutant EGFR comprises a combination of mutations, wherein the combination is selected from L858R / L844V, L858R / T790M, L858R / T790M / I941R, L858R / C797S, L858R / T790M / C797S, and L858R / T790M. In other embodiments, the mutant EGFR comprises a combination of mutations, wherein the combination is selected from L858R / T790M, L858R / T790M / I941R, L858R / C797S, L858R / T790M / C797S, and L858R / T790M.

[0090] In some embodiments, the compound of the present disclosure is combined with a second active agent, wherein the second active agent prevents EGFR dimer formation, and can regulate (for example, suppress or reduce) the activity of the EGFR containing one or more mutations. In some embodiments, the mutant EGFR contains one or more mutations selected from T790M, L718Q, L844V, V948R, L858R, I941R and C797S. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from L858R / L718Q, L858R / L844V, L858R / T790M, L858R / T790M / I941R, L858R / C797S, L858R / T790M / C797S and L858R / T790M / L718Q. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from L858R / L844V, L858R / T790M, L858R / T790M / I941R, L858R / C797S, L858R / T790M / C797S and L858R / T790M. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from L858R / T790M, L858R / T790M / I941R, L858R / C797S, L858R / T790M / C797S and L858R / T790M. In some embodiments, the second activating agent that prevents the formation of EGFR dimers is an antibody. In further embodiments, the second activating agent that prevents the formation of EGFR dimers is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib, or erlotinib.

[0091] In some embodiments, the compounds of the present disclosure are capable of modulating (eg, inhibiting or reducing) the activity of EGFR containing one or more mutations, but do not affect the activity of wild-type EGFR.

[0092] In other embodiments, the compound of the present disclosure is combined with a second active agent, wherein the second active agent prevents EGFR dimer formation, can regulate (e.g., inhibit or reduce) the activity of EGFR containing one or more mutations, but does not affect the activity of wild-type EGFR. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

[0093] Modulating EGFRs containing one or more mutations, such as those described herein, rather than wild-type EGFR, provides methods for treating, preventing, or ameliorating diseases including, but not limited to, cancer and metastasis, inflammation, arthritis, systemic lupus erythematosus, skin-related disorders, lung disorders, cardiovascular disease, ischemia, neurodegenerative disorders, liver disease, gastrointestinal disorders, viral and bacterial infections, central nervous system disorders, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal cord injury, and peripheral neuropathy.

[0094] In some embodiments, inhibition of EGFR activity is achieved by IC 50 to measure.

[0095] In some embodiments, inhibition of EGFR activity is achieved by EC 50 to measure.

[0096] In some embodiments, the inhibition of EGFR by the compounds of the present disclosure can be measured by biochemical analysis. By way of illustrative and non-limiting example, uniform time-resolved fluorescence (HTRF) assays can be used to determine the inhibition of EGFR activity using the conditions and experimental parameters disclosed herein. For example, the HTRF assay can, for example, employ a substrate (e.g., biotin-Lck-peptide substrate) concentration of about 1 μM; an EGFR (mutant or WT) concentration from about 0.2 nM to about 40 nM; and an inhibitor concentration from about 0.000282 μM to about 50 μM. The compounds of the present disclosure screened under these conditions can, for example, exhibit an IC from about 1 nM to> 1 μM, from about 1 nM to about 400 nM, from about 1 nM to about 150 nM, from about 1 nM to about 75 nM, from about 1 nM to about 40 nM, from about 1 nM to about 25 nM, from about 1 nM to about 15 nM, or from about 1 nM to about 10 nM. 50 In certain embodiments, compounds of the present disclosure screened for inhibition of EGFR having a mutation or combination of mutations selected from L858R / T790M, L858R and T790M under the above conditions can, for example, exhibit an IC from about 1 nM to >1 μM, from about 1 nM to about 400 nM, from about 1 nM to about 150 nM, from about 1 nM to about 75 nM, from about 1 nM to about 40 nM, from about 1 nM to about 25 nM, from about 1 nM to about 15 nM, or from about 1 nM to about 10 nM. 50 value.

[0097] In some embodiments, the compounds of the present disclosure bind to the allosteric site in EGFR. In some embodiments, the compounds of the present disclosure interact with at least one amino acid residue of the epidermal growth factor receptor (EGFR) selected from Lys745, Leu788 and Ala 743. In other embodiments, the compounds of the present disclosure interact with at least one amino acid residue of the epidermal growth factor receptor (EGFR) selected from Cys755, Leu777, Phe856 and Asp855. In other embodiments, the compounds of the present disclosure interact with at least one amino acid residue of the epidermal growth factor receptor (EGFR) selected from Met766, Ile759, Glu762 and Ala763. In other embodiments, the compounds of the present disclosure interact with at least one amino acid residue of the epidermal growth factor receptor (EGFR) selected from Lys745, Leu788, and Ala 743, at least one amino acid residue of the epidermal growth factor receptor (EGFR) selected from Cys755, Leu777, Phe856, and Asp855, and at least one amino acid residue of the epidermal growth factor receptor (EGFR) selected from Met766, Ile759, Glu762, and Ala763. In other embodiments, the compounds of the present disclosure do not interact with any amino acid residue of the epidermal growth factor receptor (EGFR) selected from Met793, Gly796, and Cys797.

[0098] In some embodiments, the present disclosure provides a compound comprising an allosteric kinase inhibitor, wherein the compound is a more effective drug-resistant EGFR mutant inhibitor relative to wild-type EGFR. In some embodiments, the drug-resistant EGFR mutant is resistant to one or more known EGFR inhibitors (including but not limited to gefitinib, erlotinib, lapatinib, HKI-272 and osimertinib).

[0099] In some embodiments, the drug-resistant EGFR mutant comprises a sensitizing mutation, such as L858R.

[0100] In some embodiments, the present disclosure provides a compound comprising an allosteric kinase inhibitor in combination with a second active agent, wherein the second active agent prevents EGFR dimer formation, wherein the compound is an inhibitor of a drug-resistant EGFR mutant that is more effective than wild-type EGFR. In some embodiments, drug-resistant EGFR mutants are resistant to one or more known EGFR inhibitors (including but not limited to gefitinib, erlotinib, lapatinib, HKI-272 and osimertinib). In some embodiments, the drug-resistant EGFR mutants include sensitizing mutations, such as L858R. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0101] In other embodiments, the present disclosure provides a compound comprising an allosteric kinase inhibitor in combination with a second active agent, wherein the second active agent prevents EGFR dimer formation, wherein the compound is more effective than one or more known EGFR inhibitors in suppressing the activity of EGFR containing one or more mutations as described herein (such as T790M, L718Q, L844V, L858R and C797S) in combination with the second active agent, wherein the known EGFR inhibitors include but are not limited to gefitinib, erlotinib, lapatinib, HKI-272 and osimertinib. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0102] In other embodiments, the present disclosure provides an allosteric kinase inhibitor compound in combination with a second active agent, wherein the second active agent prevents EGFR dimer formation, wherein the compound in combination with the second active agent is less effective than one or more known EGFR inhibitors in inhibiting the activity of wild-type EGFR, the known EGFR inhibitors including but not limited to gefitinib, erlotinib, lapatinib, HKI-272 and osimertinib. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0103] The potency of the inhibitor can be determined by EC 50 Having a lower EC as determined under substantially similar conditions 50 Compounds with higher EC 50 In some embodiments, substantially similar conditions include determining the level of EGFR-dependent phosphorylation in vitro or in vivo (e.g., in 3T3 cells expressing wild-type EGFR, mutant EGFR, or any fragment thereof).

[0104] The potency of the inhibitor can be determined by IC 50 Having a lower IC as determined under substantially similar conditions 50 Compounds with higher IC 50 In some embodiments, substantially similar conditions include determining the level of EGFR-dependent phosphorylation in vitro or in vivo (e.g., in 3T3 cells expressing wild-type EGFR, mutant EGFR, or any fragment thereof).

[0105] EGFR sensitizing mutations include, but are not limited to, L858R, G719S, G719C, G719A, and / or L861Q. Drug-resistant EGFR mutants may have, but are not limited to, drug-resistant mutations including T790M, T854A, L718Q, C797S, or D761Y.

[0106] The selectivity between wild-type EGFR and the EGFR containing one or more mutations as described herein can also be measured using cell proliferation assays, wherein cell proliferation depends on kinase activity.For example, the wild-type EGFR (such as VIII containing WT EGFR kinase domains) of suitable form can be used for transfection of mouse Ba / F3 cells, or the Ba / F3 cells of L858R / T790M, L858R / T790M / L718Q, L858R / C797S, L858R / T790M / C797S or L858R / T790M / I941R can be measured for proliferation under a series of inhibitor concentrations (10 μM, 3 μM, 1.1 μM, 330 nM, 110 nM, 33 nM, 11 nM, 3 nM, 1nM), and EC is calculated. 50 .

[0107] Another method to measure the effect on EGFR activity is to measure EGFR phosphorylation. Wild-type or mutant (L858R / T790M, L858R / C797S, L858R / T790M / C797S, L858R / T790M / I941R, or L858R / T790M / L718Q) EGFR can be transfected into NIH-3T3 cells (which do not normally express endogenous EGFR), and the ability of the inhibitor (using the above concentrations) to inhibit EGFR phosphorylation can be measured. Cells are exposed to increasing concentrations of inhibitor for 6 hours and stimulated with EGF for 10 minutes. The effect on EGFR phosphorylation is detected by Western blotting using a phospho-specific (Y1068) EGFR antibody.

[0108] On the other hand, the present disclosure provides a method for inhibiting epidermal growth factor receptor (EGFR), the method comprising administering an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof to a subject in need. In some embodiments, the method further comprises administering a second active agent, wherein the second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

[0109] In another aspect, provided herein are methods for treating or preventing a disease, comprising administering an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof to a subject in need thereof. In some embodiments, the disease is mediated by a kinase. In further embodiments, the kinase comprises a mutated cysteine ​​residue. In further embodiments, the mutated cysteine ​​residue is located at or near a position equivalent to Cys 797 in EGFR, including such positions in Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4 (ErbB4), Itk, Tec, and Txk. In some embodiments, the method further comprises administering a second active agent, wherein the second active agent prevents dimer formation of the kinase. In some embodiments, the second active agent that prevents kinase dimer formation is an antibody. In further embodiments, the second active agent prevents EGFR dimer formation. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib, or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0110] In some embodiments, the disease is mediated by EGFR (e.g., EGFR plays a role in the initiation or development of the disease). In some embodiments, the disease is mediated by Her kinase. In further embodiments, the Her kinase is HER1, HER2, or HER4.

[0111] In certain embodiments, the disease is resistant to known EGFR inhibitors, including but not limited to gefitinib, erlotinib or osimertinib. In certain embodiments, a diagnostic test is performed to determine whether the disease is associated with an activating mutation in EGFR. In certain embodiments, a diagnostic test is performed to determine whether the disease is associated with an EGFR carrying an activating mutation and / or a drug-resistant mutation. Activating mutations include but are not limited to L858R, G719S, G719C, G719A, L718Q and / or L861Q. Drug-resistant EGFR mutants may have, but are not limited to, drug-resistant mutations including T790M, T854A, L718Q, C797S or D761Y. Diagnostic tests may include sequencing, pyrosequencing, PCR, RT-PCR or similar analytical techniques known to those skilled in the art that can detect nucleotide sequences.

[0112] In certain embodiments, the disease is cancer or a proliferative disease.

[0113] In further embodiments, the disease is lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, brain cancer, kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer, stomach cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, hepatocellular carcinoma, papillary renal carcinoma, head and neck squamous cell carcinoma, leukemia, lymphoma, myeloma, or a solid tumor. In further embodiments, the disease is lung cancer, breast cancer, glioma, squamous cell carcinoma, or prostate cancer. In yet further embodiments, the disease is non-small cell lung cancer.

[0114] In certain embodiments, the disease is resistant to known EGFR inhibitors, including but not limited to gefitinib, erlotinib or osimertinib. In certain embodiments, a diagnostic test is performed to determine whether the disease is associated with an activating mutation in EGFR. In certain embodiments, a diagnostic test is performed to determine whether the disease is associated with an EGFR carrying an activating mutation and / or a drug-resistant mutation. Activating mutations include but are not limited to L858R, G719S, G719C, G719A, L718Q and / or L861Q. Drug-resistant EGFR mutants may have, but are not limited to, drug-resistant mutations including T790M, T854A, L718Q, C797S or D761Y. Diagnostic tests may include sequencing, pyrosequencing, PCR, RT-PCR or similar analytical techniques known to those skilled in the art that can detect nucleotide sequences.

[0115] In yet another aspect, provided herein are methods for treating kinase-mediated disorders comprising administering to a subject in need thereof an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an inhibitor of HER1, HER2, or HER4. In other embodiments, an additional therapeutic agent is administered to the subject. In other embodiments, the compound and the additional therapeutic agent are administered simultaneously or sequentially.

[0116] In another aspect, the present disclosure provides a method for treating a kinase-mediated condition, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and a second active agent, wherein the second active agent prevents EGFR dimer formation. In some embodiments, the compound is an inhibitor of HER1, HER2, or HER4. In other embodiments, an additional therapeutic agent is administered to the subject. In other embodiments, the compound, the second active agent that prevents EGFR dimer formation, and the additional therapeutic agent are administered simultaneously or sequentially. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib, or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0117] In other embodiments, the disease is cancer. In further embodiments, the cancer is lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, brain cancer, kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer, stomach cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, hepatocellular carcinoma, papillary renal carcinoma, head and neck squamous cell carcinoma, leukemia, lymphoma, myeloma, or solid tumor. In further embodiments, the disease is lung cancer, breast cancer, glioma, squamous cell carcinoma, or prostate cancer. In yet further embodiments, the disease is non-small cell lung cancer.

[0118] In another aspect, provided herein is a method of treating cancer, wherein the cancer cells comprise activated EGFR, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof.

[0119] In another aspect, provided herein is a method for treating cancer (wherein the cancer cell comprises activated EGFR), comprising administering an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof and a second active agent to a subject in need thereof, wherein the second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

[0120] In certain embodiments, the EGFR activation is selected from the group consisting of EGFR mutation, EGFR amplification, EGFR expression, and ligand-mediated EGFR activation.

[0121] In a further embodiment, the EGFR mutation is selected from G719S, G719C, G719A, L858R, T790M and L861Q.

[0122] In yet another aspect, provided herein are methods of treating cancer in a subject, wherein the subject is determined to be in need of EGFR inhibition to treat the cancer, the method comprising administering to the subject an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof.

[0123] In certain embodiments, the subject determined to be in need of EGFR inhibition is resistant to known EGFR inhibitors, including but not limited to gefitinib, erlotinib, or osimertinib. In certain embodiments, a diagnostic test is performed to determine whether the subject has an activating mutation in EGFR. In certain embodiments, a diagnostic test is performed to determine whether the subject has an EGFR carrying an activating mutation and / or a drug-resistant mutation. Activating mutations include but are not limited to L858R, G719S, G719C, G719A, L718Q, and / or L861Q. Drug-resistant EGFR mutants may have, but are not limited to, drug-resistant mutations including T790M, T854A, L718Q, C797S, or D761Y. Diagnostic tests may include sequencing, pyrosequencing, PCR, RT-PCR, or similar analytical techniques known to those skilled in the art that can detect nucleotide sequences.

[0124] In one aspect, provided herein are methods for preventing a subject from developing resistance to known EGFR inhibitors (including but not limited to gefitinib, erlotinib, or osimertinib), comprising administering to a subject in need thereof an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof.

[0125] In another aspect, provided herein is a method for preventing the disease from developing resistance to known EGFR inhibitors (including but not limited to gefitinib, erlotinib or osimertinib), comprising administering an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof and a second activating agent to a subject in need thereof, wherein the second activating agent prevents EGFR dimer formation. In some embodiments, the second activating agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second activating agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second activating agent that prevents EGFR dimer formation is cetuximab.

[0126] In one embodiment of the methods disclosed herein, the subject is a human.

[0127] In another aspect, the present disclosure provides a compound disclosed herein, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for treating or preventing a disease in which EGFR plays a role.

[0128] In one aspect, provided herein are methods of treating or preventing a condition selected from the group consisting of autoimmune diseases, inflammatory diseases, proliferative and hyperproliferative diseases, immune-mediated diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cardiovascular diseases, hormone-related diseases, allergies, asthma, and Alzheimer's disease. In other embodiments, the condition is selected from a proliferative disorder and a neurodegenerative disorder.

[0129] One aspect of the present disclosure provides compounds that can be used to treat diseases, conditions, and disorders characterized by excessive or abnormal cell proliferation. Such diseases include, but are not limited to, proliferative or hyperproliferative diseases and neurodegenerative diseases. Examples of proliferative and hyperproliferative diseases include, but are not limited to, cancer. The term "cancer" includes, but is not limited to, the following cancers: breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, stomach cancer, skin cancer, keratoacanthoma, lung cancer, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, colorectal cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, liver cancer and biliary tract cancer, kidney cancer, bone marrow disorders, lymphoid disorders, hairy cell cancer, oral cavity and pharyngeal (mouth) cancer, lip cancer, tongue cancer, mouth cancer, pharyngeal cancer, small intestine cancer, colon cancer, rectum cancer, large intestine cancer, rectal cancer, brain cancer and central nervous system cancer, chronic myeloid leukemia (CML) and leukemia. The term "cancer" includes, but is not limited to, myeloma, lymphoma, or a cancer selected from gastric cancer, renal cancer, head and neck cancer, oropharyngeal cancer, non-small cell lung cancer (NSCLC), endometrial cancer, liver cancer, non-Hodgkin's lymphoma, and lung cancer.

[0130] The term "cancer" also refers to any cancer caused by the proliferation of malignant cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas, etc. For example, cancer includes, but is not limited to, mesothelioma, leukemias, and lymphomas, such as cutaneous T-cell lymphoma (CTCL), non-cutaneous peripheral T-cell lymphomas, lymphomas associated with human T-cell lymphotropic virus (HTLV) (such as adult T-cell leukemia / lymphoma (ATLL)), B-cell lymphomas, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, lymphomas and multiple myeloma, non-Hodgkin's lymphoma, acute lymphoid leukemia (ALL), chronic lymphoid leukemia (CLL), Hodgkin's lymphoma, Burkitt's lymphoma, adult T-cell leukemia lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), or hepatocellular carcinoma. Further examples include myelodysplastic syndrome, solid tumors in children (such as brain tumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors and soft tissue sarcomas), common solid tumors in adults (such as head and neck cancer (e.g., oral cancer, laryngeal cancer, nasopharyngeal cancer and esophageal cancer), genitourinary cancer (e.g., prostate cancer, bladder cancer, kidney cancer, uterine cancer, ovarian cancer, testicular cancer), lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), breast cancer, pancreatic cancer, melanoma and other skin cancers, stomach cancer, brain tumors, tumors associated with Gorlin's syndrome (e.g., medulloblastoma, meningioma, etc.) and liver cancer). Additional exemplary forms of cancer that can be treated by the subject compounds include, but are not limited to, bone cancer or smooth muscle cancer, stomach cancer, small intestine cancer, rectal cancer, salivary gland cancer, endometrial cancer, adrenal cancer, anal cancer, rectal cancer, parathyroid cancer and pituitary cancer.

[0131] Additional cancers for which the compounds described herein can be used for prevention, treatment, and research include, for example, colon cancer, familial adenomatous polyposis carcinoma, and hereditary non-polyposis colorectal cancer, or melanoma. In addition, cancers include, but are not limited to, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, thyroid cancer (medullary and papillary thyroid cancer), kidney cancer, renal parenchymal cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, testicular cancer, urinary tract cancer, melanoma, brain tumors (such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral neuroectodermal tumor), gallbladder cancer, bronchogenic carcinoma, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma. In one aspect of the present disclosure, the present disclosure provides the use of one or more compounds of the present disclosure in the manufacture of a medicament for treating cancer, including but not limited to the various types of cancer disclosed herein.

[0132] In some embodiments, the compounds of the present disclosure can be used to treat cancers such as colorectal cancer, thyroid cancer, breast cancer, and lung cancer; and myeloproliferative disorders such as polycythemia vera, thrombocythemia, myeloid metaplasia with myelofibrosis, chronic myeloid leukemia, chronic myelomonocytic leukemia, hypereosinophilic syndrome, juvenile myelomonocytic leukemia, and systemic mast cell disease. In some embodiments, the compounds of the present disclosure can be used to treat hematopoietic disorders, specifically acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute promyelocytic leukemia, and acute lymphoblastic leukemia (ALL).

[0133] The term "cancer cell" as provided herein includes cells affected by any of the conditions described above.

[0134] The present disclosure also provides methods for treating or preventing cell proliferative diseases such as hyperplasia, dysplasia, and precancerous lesions. Dysplasia is the earliest form of precancerous lesion that a pathologist can identify through biopsy. The subject compounds can be administered to prevent the hyperplasia, dysplasia, or precancerous lesion from continuing to expand or becoming cancerous. Examples of precancerous lesions may occur in the skin, esophageal tissue, breast, and cervical intraepithelial tissue.

[0135] Examples of neurodegenerative diseases include, but are not limited to, adrenoleukodystrophy (ALD), Alexander disease, Alper disease, Alzheimer's disease, amyotrophic lateral sclerosis (Lou Gehrig's disease), ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, corticobasal degeneration, Creutzfeldt-Jakob disease, familial fatal insomnia, frontotemporal lobar degeneration, Huntington's disease, HIV-associated dementia, Kennedy disease, Clapper's disease, dementia with Lewy bodies, neuroborreliosis, equine Chateau-Joseph disease (spinocerebellar ataxia type 3), multiple system atrophy, multiple sclerosis, narcolepsy, Niemann-Pick disease, Parkinson's disease, Perelman-Merzheimer disease, Pick's disease, primary lateral sclerosis, prion diseases, progressive supranuclear palsy, Refsum's syndrome, Sandhoff's disease, Scheldt disease, subacute combined degeneration of the spinal cord due to pernicious anemia, Spielmeyer-Vogt-Sjögren-Batten disease (also known as Batten disease), spinocerebellar ataxias (various types with varying features), spinal muscular atrophy, supranuclear palsy syndrome, tabes dorsalis, and toxic encephalopathy.

[0136] Another aspect of the present disclosure provides a method for treating or alleviating the severity of a disease selected from a proliferative disease or a hyperproliferative disease or a neurodegenerative disease, comprising administering an effective amount of a compound or a pharmaceutically acceptable composition comprising a compound to a subject in need. In other embodiments, the method further comprises administering a second active agent, wherein the second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0137] The activity of the compounds and compositions of the present disclosure as EGFR kinase inhibitors can be determined in vitro, in vivo, or in cell lines. In vitro assays include assays to determine the kinase activity or ATPase activity of kinases that inhibit activation. Alternative in vitro assays can quantify the ability of inhibitors to bind to protein kinases and can be measured by radiolabeling the inhibitor prior to binding, isolating the inhibitor / kinase complex and determining the amount of radiolabeled binding, or by running competition experiments in which the new inhibitor is incubated with a kinase that binds a known radioligand. Detailed conditions for determining compounds used as inhibitors of various kinases in the present disclosure are listed in the following examples.

[0138] In accordance with the foregoing, the present disclosure further provides a method for preventing or treating any of the above-mentioned diseases or conditions in a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and optionally a second active agent, wherein the second active agent prevents EGFR dimer formation. For any of the above uses, the required dosage will vary depending on the mode of administration, the specific condition to be treated, and the desired effect.

[0139] In other embodiments, the compound and a second agent that prevents EGFR dimer formation are administered simultaneously or sequentially.

[0140] Administration / dosage / formulation Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents; solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and fragrances.

[0141] Injectable preparations (e.g., sterile injectable aqueous or oily suspensions) can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be sterile injectable solutions, suspensions, or emulsions contained in a nontoxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, USP, and isotonic sodium chloride solution. In addition, sterile, fixed oils are commonly used as solvents or suspending media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injectable preparations.

[0142] To prolong the effect of a drug, it is often necessary to slow the absorption of a drug injected subcutaneously or intramuscularly. This can be achieved by using a liquid suspension of a poorly water-soluble crystalline or amorphous substance. The absorption rate of a drug depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0143] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the disclosed compounds with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0144] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose (milk sugar) and high molecular weight polyethylene glycols.

[0145] The active compound can also be in the form of a microencapsulation with one or more excipients as described above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings, controlled release coatings and other coatings known in the field of pharmaceutical preparations. In such solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. Common practice is that such dosage forms can also include additional substances in addition to the inert diluent, such as tableting lubricants and other tableting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form can also include a buffer.

[0146] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. As may be desired, the active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any desired preservatives or buffers. Ophthalmic preparations, eye drops, ophthalmic ointments, powders, and solutions are also encompassed within the scope of the present disclosure.

[0147] The ointments, pastes, creams and gels may contain, in addition to the active compounds of this disclosure, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0148] Powders and sprays can contain, in addition to a compound of this disclosure, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays can also contain customary propellants such as chlorofluorohydrocarbons.

[0149] Transdermal patches have the advantage of increasing the controlled delivery of compounds to the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. Rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0150] According to the treatment methods of the present invention, a therapeutically effective amount of a compound of the present invention is administered to a subject (such as a human or other animal) in an amount and for a time required to achieve the desired result to treat or prevent the subject's condition. As used herein, the term "therapeutically effective amount" of a compound of the present invention means an amount of the compound sufficient to alleviate the symptoms of the subject's condition. As is well known in the medical field, a therapeutically effective amount of a compound of the present invention will have a reasonable benefit / risk ratio applicable to any medical treatment.

[0151] In general, the compounds of the present disclosure will be administered alone or in combination with one or more therapeutic agents in any commonly used and acceptable manner known in the art. A therapeutically effective amount may vary greatly depending on the severity of the disease, the age and relative health of the subject, the efficacy of the compound used, and other factors. In general, a daily dose of about 0.03 to 2.5 mg / kg according to body weight can systematically obtain satisfactory results. For larger mammals (e.g., humans), the indicated daily dose is within the range of about 0.5 mg to about 100 mg, and can be conveniently administered, for example, in divided doses (up to four times a day) or in a sustained-release form. A unit dosage form suitable for oral administration comprises from about 1 to 50 mg of active ingredient.

[0152] In certain embodiments, the therapeutic amount or dosage of the compound of the present disclosure can be from about 0.1 mg / Kg to about 500 mg / Kg, or from about 1 to about 50 mg / Kg. Generally speaking, the treatment regimen according to the present disclosure includes administering from about 10 mg to about 1000 mg of the compound of the present disclosure to a patient in need of such treatment in a single dose or multiple doses per day. The therapeutic amount or dosage will also vary depending on the route of administration and the possibility of co-use with other drugs.

[0153] When the subject's condition improves, a maintenance dose of a compound, composition, or combination of the present invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, depending on the symptoms, to a level that maintains the improved condition; treatment should be discontinued when symptoms are alleviated to the desired level. However, patients may require long-term intermittent treatment upon any recurrence of disease symptoms.

[0154] However, it will be understood that the total daily usage of the compounds and compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the specific inhibitory dose will depend on a variety of factors, including the condition being treated and the severity of the condition; the activity of the specific compound employed; the specific composition employed; the patient's age, weight, general health, sex, and diet; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of treatment; drugs used in combination with or concurrently with the specific compound employed; and similar factors well known in the medical art.

[0155] The present disclosure also provides a pharmaceutical combination, such as a kit, comprising a) a first agent, which is a compound of the present disclosure as disclosed herein, in free form or a pharmaceutically acceptable salt, and b) at least one co-agent. The kit may comprise instructions for its administration.

[0156] In certain embodiments, these compositions optionally further comprise one or more additional therapeutic agents.For example, agents that prevent EGFR dimer formation, chemotherapeutic agents, or other anti-proliferative agents can be combined with the compounds of the present disclosure to treat proliferative diseases and cancer.

[0157] Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers; alumina; aluminum stearate; lecithin; serum proteins such as human serum albumin; buffer substances such as phosphate, glycine, sorbic acid, or potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes such as protamine sulfate; disodium hydrogen phosphate; potassium hydrogen phosphate; sodium chloride; zinc salts; colloidal silicon dioxide; magnesium trisilicate; polyvinyl pyrrolidone; polyacrylates; waxes; polyethylene polyoxypropylene block polymers; lanolin; sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; and phosphate buffered saline. In addition, at the discretion of the formulator, non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavorings and fragrances, preservatives and antioxidants may also be present in the composition. The protein kinase inhibitor or a pharmaceutically acceptable salt thereof can be formulated into a pharmaceutical composition for animal or human administration. These pharmaceutical compositions are other embodiments of the present disclosure and comprise an amount of a protein inhibitor effective to treat or prevent a protein kinase-mediated disorder and a pharmaceutically acceptable carrier.

[0158] medicine box In one aspect, the present invention provides a kit comprising a compound capable of inhibiting kinase activity selected from one or more compounds disclosed herein, or a pharmaceutically acceptable salt thereof, and instructions for treating cancer. In certain embodiments, the kit further comprises components for performing a test to determine whether a subject has an activating and / or drug-resistant mutation in EGFR.

[0159] In another aspect, the present disclosure provides a kit comprising a compound capable of inhibiting EGFR activity selected from the compounds disclosed herein or a pharmaceutically acceptable salt thereof.

[0160] In another aspect, the present disclosure provides a kit comprising a compound capable of inhibiting kinase activity, the compound being selected from one or more compounds disclosed herein or a pharmaceutically acceptable salt thereof; a second active agent, wherein the second active agent prevents the formation of EGFR dimers; and instructions for treating cancer. In certain embodiments, the kit further comprises components for conducting tests to determine whether a subject has an activating and / or drug resistance mutation in EGFR. In some embodiments, the second active agent that prevents the formation of EGFR dimers is an antibody. In a further embodiment, the second active agent that prevents the formation of EGFR dimers is cetuximab, trastuzumab, or panitumumab. In a further embodiment, the second active agent that prevents the formation of EGFR dimers is cetuximab.

[0161] In another aspect, the present disclosure provides a drug kit comprising a compound capable of inhibiting EGFR activity selected from the compounds disclosed herein or a pharmaceutically acceptable salt thereof and a second active agent, wherein the second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab or panitumumab. In a further embodiment, the second active agent that prevents EGFR dimer formation is cetuximab. In one embodiment, the second active agent is an ATP-competitive EGFR inhibitor. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP-competitive EGFR inhibitor is osimertinib.

[0162] The present disclosure is further illustrated by the following examples and synthetic schemes, which should not be construed as limiting the scope or spirit of the present invention to the specific procedures described herein. It should be understood that the examples are provided to illustrate certain embodiments and are not intended to limit the scope of the present disclosure. It should be further understood that it would be conceivable for those skilled in the art to make various other embodiments, modifications, and equivalents without departing from the spirit of the present disclosure and / or the scope of the appended claims. Example

[0163] The present invention is further illustrated by the following examples, which should in no way be construed as further limiting. Unless otherwise indicated, the practice of this disclosure will employ conventional techniques of organic synthesis, cell biology, cell culture, and molecular biology, which are within the skill of the art.

[0164] abbreviation ACN acetonitrile DCM dichloromethane DIEA Diisopropylethylamine DMF dimethylformamide DMSO dimethyl sulfoxide EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EtOAc HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate HBTU 3-[Bis(dimethylamino)methyl]-3H-benzotriazole-1-oxide hexafluorophosphate MeOH methanol TFA trifluoroacetic acid RT Room temperature THF Tetrahydrofuran

[0165] Example 1: Synthesis procedure Scheme 1. Synthesis of Compound 001 (E)-4-chloro-2-(2-ethoxyvinyl)-6-fluorobenzoic acid methyl ester Methyl 2-bromo-4-chloro-6-fluorobenzoate (1.00 g, 3.74 mmol), ( E A mixture of 2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (741 mg, 3.74 mmol), KCO (1.55 mg, 11.22 mmol), water (3 mL), and dioxane (30 mL) was placed in a sealed vial, degassed, and purged with nitrogen three times. Pddba (171 mg, 0.187 mmol) and tricyclohexylphosphine (209 mg, 0.748 mmol) were added, and the vial was resealed, refilled with nitrogen, and heated to 80°C for 20 minutes. After cooling, the reaction was poured into saturated brine (150 mL) and extracted with EtOAc (3 × 100 mL). The combined organic extracts were washed with water and saturated brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with 0-15% EtOAc / hexanes to provide the title compound (674 mg, 70%). 1 H NMR (500 MHz, DMSO- d 6) δ ppm 7.55 (d, 1H) 7.45 (d, 1H) 7.29(dd, 1H) 5.76 (d, 1H) 3.93 (q, 2H) 3.99 (s, 3H)1.25 (t, 3H); MS ( m / z ): [M+1] + ,259.24.

[0166] (E)-3-(2-ethoxyvinyl)-5-fluoro-4'-(1-methylpiperidin-4-yl)-[1,1'-biphenyl]-4-carboxylic acid ester Will( E A mixture of methyl 4-chloro-2-(2-ethoxyvinyl)-6-fluorobenzoate (348 mg, 1.34 mmol), Pd(OAc)2 (30 mg, 0.134 mmol), 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine (813 mg, 2.69 mmol), S-Phos (110 mg, 0.269 mmol), Cs2CO3 (1.75 g, 5.38 mmol) and CuCl (27 mg, 0.269 mmol) in DMF (20 mL) was placed in a sealed vial, degassed and refilled with N2 three times. The mixture was stirred at 80° C. for 1 h, cooled, filtered, and directly purified by reverse phase HPLC eluting with 0-80% ACN / H 2 O (TFA modifier, 0.0375%) to afford the title compound (446 mg, 65%) as a TFA salt. 1 H NMR (500 MHz, DMSO- d 6) δ ppm 9.45 (br s, 1H) 7.77 (d, 2H) 7.67 (d, 1H)7.47 (d, 1H) 7.39 (d, 1H) 7.36 (d,2H) 5.86 (d, 1H) 3.95 (q, 2H) 3.89 (s, 3H)3.54 (d, 2H) 3.10 (m, 2H) 2.87 (m, 1H) 2.84 (d, 3H) 2.05 (d, 2H) 1.87 (m,2H); MS ( m / z ): [M+1] + , 398.17.

[0167] Lithium (E)-3-(2-ethoxyvinyl)-5-fluoro-4'-(1-methylpiperidin-4-yl)-[1,1'-biphenyl]-4-carboxylate Will( EA mixture of methyl 3-(2-ethoxyvinyl)-5-fluoro-4'-(1-methylpiperidin-4-yl)-[1,1'-biphenyl]-4-carboxylate (TFA salt, 440 mg, 0.86 mmol) and LiOH-H2O (100 mg, 2.43 mmol) in THF (3 mL), MeOH (3 mL) and water (3 mL) was heated to 50°C for 5 hours. The solvent was removed under reduced pressure and the residue was dried in vacuo at 60°C overnight and used without further purification. MS ( m / z ): [M+1] + , 384.14.

[0168] 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(8-fluoro-6-(4-(1-methylpiperidin-4-yl) ethyl 1-oxoisoquinolin-2(1H)-yl)acetate A solution of lithium (E)-3-(2-ethoxyvinyl)-5-fluoro-4'-(1-methylpiperidin-4-yl)-[1,1'-biphenyl]-4-carboxylate (214 mg, 0.55 mmol) in DMF (1 mL) was treated with HBTU (229 mg, 0.61 mmol), and the mixture was stirred at room temperature for 1 hour. A solution of ethyl 2-amino-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate trifluoroacetic acid ethyl ester (240 mg, 0.74 mmol) in DMF (1 mL) was added, and the reaction mixture was stirred at room temperature for 10 minutes. Most of the DMF was evaporated under a stream of N2, and the residue was partitioned between water (30 mL) and DCM (30 mL). The aqueous layer was further extracted with DCM (2 × 30 mL), and the combined organic extracts were dried over Na2SO4 and filtered. The filtrate was treated with 4N HCl in dioxane (2 mL) and after standing for 5 minutes, the solvent was removed under reduced pressure. The residue was purified by reverse phase HPLC (0-80% ACN / H2O (TFA modifier)) to give the title compound (196 mg, 55%) as a TFA salt. 1 HNMR (500 MHz, DMSO- d6) δ ppm 9.56 (br s, 1 H) 8.74 (br s, 1 H) 7.86 (d, 1 H)7.83 (d, 2H) 7.64 (dd, 1 H)7.52 (d, 1 H) 7.40 (d, 2 H) 6.78 (dd, 1 H) 6.53(s, 1 H) 4.24 (q, 2 H) 4.21 (m, 2H) 3.55 (d, 2H) 3.10 (m, 2 H) 2.89 (m, 2H)2.85 (d, 3H) 2.80 (m, 1H) 2.56 (m,2 H) 2.06 (d, 2 H) 1.88 (m, 2 H) 1.21(t, 3H); MS ( m / z ): [M+1] + , 528.99.

[0169] 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(8-fluoro-6-(4-(1-methylpiperidin-4-yl) phenyl)-1-oxoisoquinolin-2(1H)-yl)acetic acid A solution of ethyl 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(8-fluoro-6-(4-(1-methylpiperidin-4-yl)phenyl)-1-oxoisoquinolin-2(1H)-yl)acetate (TFA salt, 192 mg, 0.3 mmol) in THF / MeOH / water (1:1:1, 4.5 mL) was treated with LiOH-H2O (62 mg, 1.5 mmol) and stirred at room temperature for 1.5 hours. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in water (7 mL) and treated with 1 N HCl to pH 2. Water was removed under reduced pressure and the residue was dried to give the title compound, which was used in the next step without further purification. MS ( m / z ): [M+1] + , 501.15.

[0170] Example 1 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(8-fluoro-6-(4-(1-methylpiperidin-1-yl)- (4-pyridin-4-yl)phenyl)-1-oxoisoquinolin-2(1H)-yl)-N-(thiazol-2-yl)acetamide A mixture of 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(8-fluoro-6-(4-(1-methylpiperidin-4-yl)phenyl)-1-oxoisoquinolin-2(1H)-yl)acetic acid (from the previous step, 0.3 mmol), HATU (125 mg, 0.33 mmol), 2-aminothiazole (45 mg, 0.45 mmol) and diisopropylethylamine (208 mL, 1.2 mmol) in DMF (3.5 mL) was stirred at room temperature for 1.5 hours. Another portion of HATUH (125 mg, 0.33 mmol) and 2-aminothiazole (45 mg, 0.45 mmol) were added, and the mixture was stirred at 50° C. for 2 hours. After cooling to room temperature, the reaction mixture was purified by reverse phase HPLC eluting with 0-80% ACN / H2O (TFA modifier) ​​to afford the title compound (110 mg, 53% yield over two steps) as a TFA salt. 1 H NMR (500 MHz, DMSO- d 6) δ ppm 9.46 (br s, 1 H) 7.86 (m,1 H) 7.85(d, 2H) 7.64 (dd, 1 H) 7.53 (d, 1 H) 7.40 (d, 2 H) 7.32 (d, 1 H)7.30 (s, 1H) 6.78 (s, 1 H) 6.72 (dd, 1 H) 4.17 (m, 2 H) 3.55 (d, 2 H) 3.10 (m, 2 H) 2.89 (m, 2H) 2.85 (s, 3H) 2.80 (m, 1H) 2.56 (m, 2 H) 2.07 (d, 2 H)1.87 (m, 2 H); MS ( m / z ): [M+1] + , 583.23.

[0171] 2-Amino-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetic acid Ethyl 2-amino-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate was prepared according to the procedure described in WO 2020 / 002487.

[0172] Compound 2 2-((R)-6-fluoro-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(8-fluoro-6-(4- (1-methylpiperidin-4-yl)phenyl)-1-oxoisoquinolin-2(1H)-yl)-N-(thiazol-2-yl)acetamide Compound 2 is in accordance with Compound1 was prepared in the same manner from lithium (E)-3-(2-ethoxyvinyl)-5-fluoro-4'-(1-methylpiperidin-4-yl)-[1,1'-biphenyl]-4-carboxylate and ethyl 2-amino-2-((R)-6-fluoro-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate. MS ( m / z ): [M+1] + , 601.26.

[0173] 2-Amino-2-((R)-6-fluoro-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetic acid Ethyl 2-amino-2-((R)-6-fluoro-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate was prepared according to the procedure described in WO 2020 / 002487.

[0174] Scheme 2 - Synthesis of Compound 3 3-Bromo-5-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylic acid methyl ester Tetrakis(triphenylphosphine)palladium(0) (533 mg, 0.462 mmol) was added to a mixture of methyl 3,5-dibromothiophene-2-carboxylate (690 mg, 2.31 mmol), K2CO3 (957 mg, 6.92 mmol), 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine (764 mg, 2.54 mmol) in toluene (10 mL) and water (1 mL). The reaction vial was evacuated to nitrogen and the mixture was stirred at 80°C for 3 days. The solvent was removed under reduced pressure and the residue was dissolved in DMSO and filtered. The filtrate was purified by reverse phase HPLC eluting with 0-80% ACN / water (TFA modifier) ​​to give the title compound (412 mg, 35%) as a TFA salt. 1 H NMR (500 MHz, DMSO- d 6) δ ppm 7.77(d, 2H) 7.75(s, 1H) 7.36 (d, 2H) 3.85 (s, 3H) 3.55 (m, 2H) 3.08 (m, 2H) 2.86(m, 1H) 2.84 (d, 3H) 2.04 (m,2H) 1.84 (m, 2H); MS ( m / z ): [M+1] + 394.01.

[0175] (E)-methyl 3-(2-ethoxyvinyl)-5-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylate Pd2(dba)3 (102 mg, 0.056 mmol) and PCy3 (63 mg, 0.223 mmol) were added to a mixture of methyl 3-bromo-5-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylate (TFA salt, 440 mg, 0.87 mmol), (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (243 mg, 1.23 mmol), K2CO3 (462 mg, 3.35 mmol) in dioxane (8 mL) and water (0.8 mL). The reaction vial was evacuated to nitrogen, and the mixture was stirred at 80°C for 1 hour. The reaction mixture was cooled, filtered, and the filtrate was purified by reverse phase HPLC eluting with 0-80% ACN / water (TFA modifier) ​​to afford the title compound (182 mg, 42%) as a TFA salt. MS ( m / z ): [M+1] + 387.18.

[0176] Lithium (E)-3-(2-ethoxyvinyl)-5-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylate A mixture of (E)-methyl 3-(2-ethoxyvinyl)-5-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylate (TFA salt, 250 mg, 0.50 mmol) and LiOH monohydrate (82 mg, 1.95 mmol) in THF / MeOH / water (1:1:1, 3.0 mL) was stirred at 60° C. for 3.5 hours. The solvent was removed under reduced pressure and the residue was dried under high vacuum overnight. This material was used directly in the next step without further purification. MS ( m / z ): [M+1] + , 372.14.

[0177] 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(2-(4-(1-methylpiperidin-4-yl)benzene ethyl)-7-oxothieno[2,3-c]pyridin-6(7H)-yl)acetate HBTU (271 mg, 0.72 mmol) was added to a suspension of the crude lithium (E)-3-(2-ethoxyvinyl)-5-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylate obtained in the previous step in DMF (3 mL) and stirred at room temperature for 1 hour. A solution of ethyl 2-amino-2-(3a,4,5,6-tetrahydrocyclopenta[c]pyrrol-1-yl)acetate (210 mg, 0.65 mmol) in DMF (1 mL) was added, and the reaction was stirred at room temperature for 1.5 hours. Most of the DMF was evaporated under a stream of N2, and the residue was partitioned between water (30 mL) and DCM (30 mL). The aqueous layer was further extracted with DCM (2 × 30 mL), and the combined organic extracts were dried over Na2SO4 and filtered. The filtrate was treated with 4N HCl in dioxane (2 mL), and after standing for 5 minutes, the solvent was removed under reduced pressure. The residue was purified by reverse phase HPLC (0-80% ACN / H2O (TFA modifier)) to give the title compound (100 mg, 32% yield over two steps) as a TFA salt. MS ( m / z ): [M+1] + ,517.19.

[0178] 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(2-(4-(1-methylpiperidin-4-yl)benzene 7-Oxothieno[2,3-c]pyridin-6(7H)-yl)acetic acid A mixture of ethyl 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(2-(4-(1-methylpiperidin-4-yl)phenyl)-7-oxothieno[2,3-c]pyridin-6(7H)-yl)acetate (TFA salt, 100 mg, 0.16 mmol), LiOH monohydrate (37 mg, 0.9 mmol), THF (1 mL), MeOH (1 mL) and water (1 mL) was stirred at room temperature for 2 hours. The organic solvent was removed under reduced pressure, water (5 mL) was added and the residue was treated with 1N HCl to a pH of 3-4. The solvent was removed under reduced pressure and the residue was dried under high vacuum overnight. This material was used in the next step without further purification. MS ( m / z ): [M+1] + , 488.92.

[0179] Compound 3 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(2-(4-(1-methylpiperidin-4- (phenyl)-7-oxothieno[2,3-c]pyridin-6(7H)-yl)-N-(thiazol-2-yl)acetamide The rough 2A mixture of -(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(2-(4-(1-methylpiperidin-4-yl)phenyl)-7-oxothieno[2,3-c]pyridin-6(7H)-yl)acetic acid (0.16 mmol), HATU (61 mg, 0.16 mmol), 2-aminothiazole (33 mg, 0.33 mmol), DIEA (83 mL) and DMF (3.5 mL) was stirred at room temperature for 1 hour. Additional HATU (61 mg), 2-aminothiazole (33 mg) and DIEA (83 mL) were added and the reaction was heated to 50°C for 1 hour. After cooling, the entire reaction mixture was purified by reverse phase HPLC eluting with 0-80% ACN / H2O (TFA modifier) ​​to give the title compound (70 mg, 77% yield over two steps) as a TFA salt. MS ( m / z ): [M+1] + , 571.02.

[0180] Compound 4 2-((R)-6-fluoro-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(2-(4-(1-methyl (4-(7H)-thiazolylpiperidin-6-yl)phenyl)-7-oxothieno[2,3-c]pyridin-4-yl)-N-(thiazol-2-yl)acetamide Compound 4 is in accordance with Compound 3 was prepared in the same manner from lithium (E)-3-(2-ethoxyvinyl)-5-fluoro-(4-(1-methylpiperidin-4-yl)phenyl)thiophene-2-carboxylate and ethyl 2-amino-2-((R)-6-fluoro-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate. MS ( m / z ): [M+1] + , 589.19.

[0181] Scheme 3 - Synthesis of Compound 5 2-(2-amino-5-bromobenzamido)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetic acid Ethyl ester A mixture of 2-amino-5-bromo-benzoic acid (500 mg, 2.31 mmol), ethyl 2-amino-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate (500 mg, 2.39 mmol), HOBt (400 mg, 2.96 mmol), EDCI (500 mg, 2.61 mmol), and diisopropylethylamine (808 mL, 4.64 mmol) in DMF (3.5 mL) was degassed and purged with N 2 , and the mixture was stirred at 20° C. under N 2 for 16 hours. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine (15 mL), dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO 2 , EtOAc:MeOH = 10 / 1) to give the title compound (890 mg, 94% yield) as a yellow solid.

[0182] 2-(6-Bromo-4-oxoquinazolin-3(4H)-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl) Ethyl acetate A mixture of ethyl 2-[(2-amino-5-bromobenzoyl)amino]-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate (500 mg, 1.23 mmol) in trimethyl orthoformate (5 mL) was degassed and purged with N2, and the mixture was stirred at 110°C under N2 for 12 hours. The reaction mixture was poured into water (50 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, EtOAc:methanol = 1:0 to 10:1) to obtain the title compound (426 mg, crude product) as a light yellow oil. 1 H NMR (400 MHz, DMSO- d 6) δ ppm 8.26 (m, 2H) 8.02 (dd,1H) 7.65 (d, 1H) 7.63 (s, 1H) 6.41 (s, 1H) 4.21 (m, 1H)4.19 (m, 1H) 4.02 (m,2H) 2.81 (m, 1H) 2.69 (m, 1H) 2.56 (m, 2H) 1.18 (t, 3H).

[0183] 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(6-(4-(1-methylpiperidin-4-yl)benzene 4-Oxoquinazolin-3(4H)-yl)acetic acid Pd(dppf)Cl2 (66 mg, 0.09 mmol) and K2CO3 (249 mg, 1.80 mmol) were added to a solution of ethyl 2-(6-bromo-4-oxo-quinazolin-3-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate (376 mg, 0.90 mmol) and 1-methyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]piperidine (299 mg, 0.99 mmol) in dioxane (4 mL) and water (0.4 mL). The mixture was stirred at 120°C under N2 for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (FA conditions, column: Waters Xbridge BEH C18 100*30mm*10mm; mobile phase: [water (NH4HCO3)-ACN]; B%: 5%-35%, 8 minutes) to give the title compound (120 mg, 27% yield) as a white solid. MS ( m / z ): [M+1] + , 484.3.

[0184] Compound 5 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(6-(4-(1-methylpiperidin-4- (4H)-yl)phenyl)-4-oxoquinazolin-3(4H)-yl)-N-(thiazol-2-yl)acetamide A mixture of 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-[6-[4-(1-methyl-4-piperidinyl)phenyl]-4-oxo-quinazolin-3-yl]acetic acid (25 mg, 0.05 mmol), thiazol-2-amine (16 mg, 0.16 mmol), diisopropylethylamine (22 mL, 0.13 mmol), and benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (27 mg, 0.06 mmol) in DMF (1 mL) was degassed and purged with N2, and the mixture was stirred under N2 at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (FA conditions, column: Phenomenex Luna C1875*30mm*3mm; mobile phase: [water(FA)-ACN]; B%: 1%-40%, 8 minutes) to give the title compound (4 mg, 14% yield) as a white solid. 1 H NMR (400 MHz, DMSO- d6) δ ppm 8.36 (d, 1H) 8.23 ​​(s, 1H)8.17 (m,1H) 7.78 (d, 1H) 7.72 (d, 2H) 7.68 (s, 1H) 7.50 (d, 1H) 7.39 (d, 2H)7.27 (d, 1H) 6.76 (s, 1H) 4.01 (m, 2H) 2.98 (d, 2H) 2.82 (m, 1H) 2.56 (m, 4H)2.30 (s,3H) 2.16 (m, 2H) 1.79 (m, 2H) 1.73 (m, 2H); MS ( m / z ): [M+1] + , 566.2.

[0185] Compound 6 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(5-fluoro-6-(4-(1-methylpiperidin-1-yl)- (4-pyridin-4-yl)phenyl)-4-oxoquinazolin-3(4H)-yl)-N-(thiazol-2-yl)acetamide Compound 6 was prepared in the same manner as Compound 5 from 6-amino-3-bromo-2-fluorobenzoic acid and ethyl 2-amino-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate. 1 H NMR (400 MHz, DMSO- d 6) δppm 8.22 (m, 1H) 7.96 (t, 1H) 7.68 (s, 1H) 7.59 (d, 1H) 7.53 (d, 2H) 7.49 (d,1H)7.38 (d, 2H) 7.27 (d, 1H) 6.68 (s, 1H) 4.01 (m, 2H) 2.91 (d, 2H) 2.82 (m,1H) 2.56 (m, 4H) 2.23 (s, 3H) 2.04 (m, 2H) 1.77 (m, 2H) 1.72 (m, 2H); MS ( m / z ): [M+1] + , 584.1.

[0186] Scheme 4 - Synthesis of Compound 7 2-(6-Bromo-4-oxoquinazolin-3(4H)-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl) Acetic acid LiOH .H2O (107 mg, 2.54 mmol) was added to a solution of ethyl 2-(6-bromo-4-oxo-quinazolin-3-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate (530 mg, 1.27 mmol) in EtOH (3 mL) and water (1 mL). The mixture was stirred at 25 °C for 12 hours and then concentrated to give a residue. Water (10 mL) was added and the pH was adjusted to 2. The mixture was filtered and concentrated under reduced pressure to give the title compound (450 mg, 91% yield) as a light yellow solid, which was used without further purification.

[0187] 2-(6-bromo-4-oxoquinazolin-3(4H)-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazole-1- base)- N -(thiazol-2-yl)acetamide HATU (234 mg, 0.616 mmol) and diisopropylethylamine (448 mL, 2.57 mmol) were added to a solution of 2-(6-bromo-4-oxo-quinazolin-3-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetic acid (200 mg, 0.513 mmol) and thiazol-2-amine (129 mg, 1.28 mmol) in DMF (2 mL). The mixture was stirred at 25°C for 16 hours. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO 2 , ethyl acetate / methanol = 1 / 0 to 10 / 1) to give the title compound (120 mg, 50% yield) as a yellow solid.

[0188] Compound 7 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(6-(4-(4-methylpiperidin-1- 4-Oxoquinazolin-3(4H)-yl)phenyl)- N -(thiazol-2-yl)acetamide Pd(dppf)Cl2 (8 mg, 0.010 mmol) and K2CO3 (29 mg, 0.212 mmol) were added to 2-(6-bromo-4-oxo-quinazolin-3-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)- NTo a solution of 1-methyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]piperazine (34 mg, 0.111 mmol) in dioxane (1 mL) and water (0.1 mL) was added thiazol-2-yl-acetamide (50 mg, 0.106 mmol). The mixture was stirred at 110°C for 12 hours. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions, column: Phenomenex Luna C18 75*30mm*3mm; mobile phase: [water (FA)-ACN]; B%: 10%-40%, 8 minutes) to give the title compound (2 mg, 3% yield) as a white solid. 1 H NMR (400 MHz, DMSO- d 6) δ ppm 8.30 (d, 1H) 8.20 (s, 1H), 8.17 (s, 1H) 8.13 (dd,1H) 7.74 (d, 1H) 7.68 (s, 1H)7.65 (d, 2H) 7.50 (d, 1H) 7.27 (d, 1H) 7.06 (d,2H) 6.76 (s, 1H) 4.01 (m, 2H) 3.20 (m, 2H) 2.81 (m, 1H) 2.23 (s, 3H) 9 protons are masked by solvent; MS ( m / z ): [M+1] + , 567.1.

[0189] Scheme 5 - Synthesis of Example 8 2-(5-Bromo-2-hydroxybenzamido)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetic acid Ethyl ester By 5-bromo-2-hydroxy-benzoic acid (500 mg, 2.30 mmol), 2-amino-2-(6,7-dihydro-5H-pyrrolo-[1,2-c] imidazole-1-yl) ethyl acetate (HCl salt, 623 mg, 2.53 mmol), HATU (1.05 g, 2.76 mmol), DIEA (1.20 mL, 6.91 mmol) mixture in DMF (5 mL) degassed and used N purge three times, then stirred at room temperature 16 hours.Reactant mixture is poured in water (30 mL), and extracted with EtOAc.Organic layer is washed with salt water (10 mL), through Na sO dry, filter and under reduced pressure concentrate.Residue is passed through column chromatography (petroleum ether / ethyl acetate=1 / 0 to 0 / 1) purifying to obtain title compound (410 mg), is yellow solid.

[0190] 2-(6-Bromo-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)-2-(6,7-dihydro-5H-pyrrolo[1, ethyl 2-c]imidazol-1-yl)acetate A mixture of ethyl 2-[(5-bromo-2-hydroxy-benzoyl)amino]-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate (50 mg, 0.122 mmol), diiodomethane (10 uL, 0.122 mmol) and Cs2CO3 (68 mg, 0.208 mmol) in DMF (1 mL) was stirred at 110 ° C for 1 hour. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (4 mL × 3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) to give the title compound (70 mg) as a brown solid.

[0191] 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(6-(4-(1-methylpiperidin-4-yl)benzene 4-Oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)acetic acid A mixture of ethyl 2-(6-bromo-4-oxo-2H-1,3-benzoxazin-3-yl)-2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)acetate (25 mg, 0.06 mmol), 1-methyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]piperidine (20 mg, 0.065 mmol), Pd(dppf)Cl2 (4.3 mg, 0.006 mmol), K2CO3 (16.4 mg, 0.012 mmol) in dioxane (1 mL) and water (0.1 mL) was degassed and purged with N2 three times and stirred at 120°C under N2 atmosphere for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (FA conditions, column: Phenomenex Luna C18 75*30mm*3um; mobile phase: [water (FA)-ACN]; B%: 5%-30%, 8 minutes) to give the title compound (18 mg, 29% yield) as a white solid. MS ( m / z ): [M+1] + 487.2.

[0192] Example 8 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-(6-(4-(1-methylpiperidin-4- 4-(4H)-yl)phenyl)-4-oxo-2H-benzo[e][1,3]oxazin-3(4H)-yl)-N-(thiazol-2-yl)acetamide A mixture of 2-(6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-1-yl)-2-[6-[4-(1-methyl-4-piperidinyl)phenyl]-4-oxo-2H-1,3-benzoxazin-3-yl]acetic acid (13 mg, 27 μmol), thiazol-2-amine (3 mg, 29 μmol), HATU (12 mg, 32 μmol), and DIEA (14 μL, 80 μmol) in DMF (1 mL) was degassed and purged with N2 three times, and stirred under N2 atmosphere at 25°C for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (neutral conditions, column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water(NH4HCO3)-ACN]; B%: 35%-65%, 8 minutes) to give the title compound (2 mg, 13% yield) as a white solid. 1 H NMR (400 MHz, DMSO- d6) δ 8.03 (d, 1H) 7.83 (dd, 1H) 7.60 (m, 3H)7.47 (d, 1H) 7.33 (d, 2H) 7.24 (br d, 1H) 7.13 (d, 1H) 6.38 (s, 1H) 5.54 (d, 1H) 5.34 (d, 1H)4.00 (m, 2H) 2.87 (d, 2H) 2.76 (m, 1H) 2.19 (s, 3H) 1.97 (m,2H) 1.74 (m, 2H)1.67 (m, 2H), 4 protons were masked by the solvent. MS ( m / z ): [M+1] + 569.3.

[0193] Example 2: HTRF-based EGFR biochemical assay EGFR biochemical activity was measured using a homogeneous time-resolved fluorescence (HTRF) assay (Cisbio). Inhibitors and DMSO standards were first dispensed into empty black, low-volume 384-well plates (Corning) using a D300 digital liquid dispenser (HP). All reactions were performed at room temperature, and solutions were added to the plates using a Multidrop Combi reagent dispenser (ThermoFisher). The reaction mixture (final volume 10 µL) contained 1 µM tyrosine kinase peptide-biotin substrate and mutant EGFR in reaction buffer (50 mM HEPES pH 7.0, 5 mM MgCl2, 1 mM MnCl2, 0.01% BSA, 2 mM TCEP, 0.1 mM NaVO4). Enzyme concentrations were adjusted to accommodate the different kinase activities (L858R 0.1 nM, L858R / T790M 0.02 nM). The enzyme reaction solution (2x concentration, 5 µL) was added to a 384-well plate containing the compound and incubated for 30 minutes. The enzyme reaction was initiated by adding 5 µL of ATP to a final concentration of 100 µM and allowed to react for 20 minutes. The reaction was quenched by adding 10 µL of phosphotyrosine antibody-Europium(III) cryptate (1:180 volume ratio) and streptavidin-XL665 (46.7 nM) in assay buffer containing EDTA. The reaction was then incubated at room temperature for 1 hour and read using a PHERAstar plate reader (excitation wavelength = 337 nm, emission wavelengths = 620 nm and 665 nm). IC 50The values ​​were determined by three replicates of the inhibition curve (11-point curve from 1.0 μM to 0.130 nM or 23-point curve from 1.0 μM to 0.130 pM) using the nonlinear least squares method fitted in GraphPad Prism 7.0d. The data are shown in Table 2 below, where "-" represents IC 50 Values ​​≥ 5 nM, “+” indicates IC 50 Values ​​<5 nM, “++” indicates IC 50 Values ​​<1 nM, and “+++” indicates IC 50 Value <0.5 nM.

[0194] Table 2. serial number <![CDATA[LRTM enzyme IC 50 (nM)]]> <![CDATA[LR enzyme IC 50 (nM)]]> 001 +++ + 002 ++ + 003 +++ + 004 +++ + 005 +++ ++ 006 +++ + 007 + - 008 +++ +

[0195] Example 3: Ba / F3 cell proliferation model EGFR mutants L858R and L858R / T790M Ba / F3 cells have been described previously (Zhou, W., et al., Nature 462, 2009, 1070-1074). All cell lines were maintained in RPMI 1640 (Cellgro; Mediatech Inc., Herndon, CA) supplemented with 10% FBS, 100 units / ml penicillin, and 100 units / ml streptomycin. The Quick Change site-directed mutagenesis kit (Stratagene; La Jolla, CA) was used according to the manufacturer's instructions. Pass Pass Site-directed mutagenesis EGFR I941R mutation. All constructs were confirmed by DNA sequencing. The constructs were delivered into the retroviral vector JP1540 using the Cre recombination system (Agilent Technologies, Santa Clara, CA). Ba / F3 cells were then infected with retrovirus according to standard protocols as previously described (Zhou et al., Nature 2009). Stable clones were obtained by selection in puromycin (2 μg / ml).

[0196] Growth and growth inhibition were assessed using the Cell Titer Glo assay (Promega, Madison, WI) and performed according to the manufacturer's instructions. The Cell Titer Glo assay is a luminescence-based method used to determine the number of viable cells based on the quantification of ATP present, which is proportional to the number of metabolically active cells present. EGFRBa / F3 cells of the same genotype were exposed to the compounds disclosed herein for 72 hours and cultured according to a previously established method (Zhou et al., Nature 2009) The number of cells used in each experiment was determined empirically. All experimental points were set up in triplicate in 384-well plates, and all experiments were repeated at least three times. Luminescence signals were detected using a spectrometer, and data were displayed graphically using GraphPad Prism version 5.0 for Windows (GraphPad software; www.graphpad.com). The curves were fitted using a nonlinear regression model with a sigmoidal dose response. The results of this assay for the compounds disclosed herein are shown in Table 3 below, where "+" indicates IC 50 Value <5 uM, "++" indicates IC 50 Value <0.5 uM, and "+++" indicates IC 50 Value <0.1 uM.

[0197] Table 3. serial number BaF3 cells L858R IC50 (uM) BaF3 cells L858R / T790M (uM) 001 ++ +++ 002 ++ ++ 003 ++ +++ 004 + ++ 005 ++ +++ 006 ++ +++ 007 + + 008 ++ +++

[0198] The disclosed subject matter is not limited to the specific embodiments and examples described herein. Indeed, various modifications of the invention other than those described herein will be apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are also within the scope of the appended claims.

[0199] All references (e.g., publications or patents or patent applications) cited herein are incorporated by reference in their entirety for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the scope of the following claims.

Claims

1. A compound of formula I: or a pharmaceutically acceptable salt thereof; in: Indicates a single bond or a double bond; A and A' are each independently CH, CR 10 , CH2, O or N; W and Z are each independently N or CR 9 ; X and Y are each independently N, CH or CR 3 ; provided that at least one of W, X, Y or Z is CH; R 1 Selected from the group consisting of 6-10 membered aryl, 5-10 membered heteroaryl, 3-10 membered heterocycloalkyl and 3-10 membered cycloalkyl, all of which are optionally substituted by one, two or three R 8 replace; Each R 2 independently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 The group consisting of NH2, CN and 3-10 membered cycloalkyl; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl or a 3-10 membered heterocycloalkyl; R 3 is independently selected at each occurrence from halogen, OR 4 NR 4 R 4 、SO2R 4 、SO2NHR 4 NHSO2R 4 、C(O)OR 4 、C(O)NHR 4 、C(O)R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, C4-C7 cycloalkenyl, C6-C 10 The group consisting of aryl, 5-6 membered heteroaryl and 4-7 membered heterocyclic group, wherein alkyl, alkenyl or alkynyl are each optionally replaced by R 4 substituted once, twice or three times, and wherein aryl, heteroaryl or heterocyclyl are each optionally replaced by R 5 Replace once, twice or thrice; R 4 is independently selected at each occurrence from H, C1-C6 alkyl, (CH2) 0-3 -(C3-C7 cycloalkyl), (CH2) 0-3 -(C4-C7 cycloalkenyl), (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 6 Replace once, twice or thrice; R 5 is independently selected at each occurrence from C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C3 alkylamine, 3-10 membered cycloalkyl, halogen, COOH, C(O)O(C1-C6 alkyl), O(CH2) 1-3 -OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, OH, CN, (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 7 Replace once, twice or thrice; R 6 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 A group consisting of NH2 or CN; R 7 is independently selected at each occurrence from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, halogen, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, SO2NH2, SO2NH(C1-C6 alkyl), SO2N(C1-C6 alkyl)2, (CH2) 1-2 -OH, C(O)(CH2) 1-2 -OH, C(O)(C1-C6 alkyl) and C(O)O(C1-C6 alkyl) substituents; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1- 4OH, S(O) 0-2 H, S(O) 0-2 The group composed of NH2 and CN; R 9 is independently selected at each occurrence from the group consisting of H, halo, C1-C3 alkyl, C1-C3 haloalkyl, and C1-C3 alkoxy; R 10 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1- 4OH, S(O) 0-2 H, S(O) 0-2 the group consisting of NH2 and CN; and n is 0, 1, 2 or 3.

2. The compound of claim 1, wherein A is CH2.

3. The compound of claim 1, wherein A is CH.

4. The compound of claim 1, wherein Indicates a single bond.

5. The compound of claim 1, wherein Represents a double bond.

6. The compound of any one of claims 1 to 5, wherein R 1 is optionally replaced by one, two or three R 8 Substituted 5-10 membered heteroaryl.

7. The compound of any one of claims 1 to 6, wherein R 1 is thiazolyl or pyridinyl, both of which are optionally substituted by one, two or three R 8 replace.

8. The compound according to any one of claims 1 to 7, wherein R 1 Selected from the group consisting of: or ; Both of which are optionally replaced by one, two or three R 8 replace.

9. The compound of any one of claims 1 to 8, wherein the compound of formula I is a compound of formula Ia: or a pharmaceutically acceptable salt thereof.

10. The compound of any one of claims 1 to 8, wherein the compound of formula I is a compound of formula Ib: or a pharmaceutically acceptable salt thereof.

11. The compound of any one of claims 1 to 8, wherein the compound of formula I is a compound of formula Id: or a pharmaceutically acceptable salt thereof.

12. The compound of any one of claims 1 to 11, wherein when X is CR 3 When , Y is CH.

13. The compound of any one of claims 1 to 11, wherein when Y is CR 3 When , then X is CH.

14. The compound of any one of claims 1-3 and 5-13, wherein A' is N.

15. The compound of any one of claims 1-3 and 5-13, wherein A' is CH.

16. The compound of any one of claims 1-4 and 6-13, wherein A' is CH2.

17. The compound of any one of claims 1-4 and 6-13, wherein A' is O.

18. The compound of any one of claims 1-8 and 12-17, wherein Z is CH or C-halo.

19. The compound of any one of claims 1 to 18, wherein R 2 is halogen or C1-C3 alkyl.

20. The compound of any one of claims 1 to 19, wherein R 3 Is one or two R 5 Substituted 6-10 membered aryl.

21. The compound of any one of claims 1 to 20, wherein R 5 is optionally replaced by R 7 4-7 membered heterocyclyl substituted once, twice or three times.

22. The compound of any one of claims 1 to 21, wherein R 3 yes or .

23. The compound of any one of claims 1 to 22, wherein R 7 It is a C1-C6 alkyl group.

24. The compound of any one of claims 1-23, wherein n is 0 or 1.

25. The compound of claim 1, wherein Indicates a single bond or a double bond; A and A' are each independently CH, CR2, O or N; W and Z are each independently CH or CR 9 ; X and Y are each independently CH or CR 3 ; R 1 is optionally replaced by one, two or three R 8 substituted 5-10 membered heteroaryl; R 2 is halogen or C1-C3 alkyl; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl or a 3-10 membered heterocycloalkyl; R 3 Is one or two R 5 substituted 6-10 membered aryl; R 5 is optionally replaced by R 7 a 4-7 membered heterocyclic group substituted once, twice or three times; R 7 is a C1-C6 alkyl group; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1- 4OH, S(O) 0-2 H, S(O) 0-2 The group composed of NH2 and CN; R 9 is independently selected at each occurrence from the group consisting of halo, C1-C3 alkyl, C1-C3 haloalkyl, and C1-C3 alkoxy; and n is 0 or 1.

26. The compound of any one of claims 1 to 25, wherein the compound of formula I is selected from the group consisting of or a group consisting of pharmaceutically acceptable salts thereof.

27. A compound of formula II: or a pharmaceutically acceptable salt thereof; A and A' are each independently CH, CR 10 or N; R 1 Selected from the group consisting of 6-10 membered aryl, 5-10 membered heteroaryl, 3-10 membered heterocycloalkyl and 3-10 membered cycloalkyl, all of which are optionally substituted by one, two or three R 8 replace; Each R 2 independently selected from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 The group consisting of NH2, CN and 3-10 membered cycloalkyl; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl or a 3-10 membered heterocycloalkyl; R 3 is independently selected at each occurrence from halogen, OR 4 NR 4 R 4 、SO2R 4 、SO2NHR 4 NHSO2R 4 、C(O)OR 4 、C(O)NHR 4 、C(O)R 4 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, C4-C7 cycloalkenyl, C6-C 10 The group consisting of aryl, 5-6 membered heteroaryl and 4-7 membered heterocyclic group, wherein alkyl, alkenyl or alkynyl are each optionally replaced by R 4 substituted once, twice or three times, and wherein aryl, heteroaryl or heterocyclyl are each optionally replaced by R 5 Replace once, twice or thrice; R 4 is independently selected at each occurrence from H, C1-C6 alkyl, (CH2) 0-3 -(C3-C7 cycloalkyl), (CH2) 0-3 -(C4-C7 cycloalkenyl), (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 6 Replace once, twice or thrice; R 5 is independently selected at each occurrence from C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C3 alkylamine, 3-10 membered cycloalkyl, halogen, COOH, C(O)O(C1-C6 alkyl), O(CH2) 1-3 -OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, OH, CN, (CH2) 0-3 -(C6-C 10 Aryl), (CH2) 0-3 -(5-6 membered heteroaryl) and (CH2) 0-3 -(4-7 membered heterocyclyl), wherein the aryl, heteroaryl or heterocyclyl are each optionally replaced by R 7 Replace once, twice or thrice; R 6 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0-2 A group consisting of NH2 or CN; R 7 is independently selected at each occurrence from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, halogen, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, SO2NH2, SO2NH(C1-C6 alkyl), SO2N(C1-C6 alkyl)2, (CH2) 1-2 -OH, C(O)(CH2) 1-2 -OH, C(O)(C1-C6 alkyl) and C(O)O(C1-C6 alkyl) substituents; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1- 4OH, S(O) 0-2 H, S(O) 0-2 The group composed of NH2 and CN; R 10 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, 3-6 membered cycloalkyl, halogen, OH, NO2, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1- 4OH, S(O) 0-2 H, S(O) 0-2 the group consisting of NH2 and CN; and n is 0, 1, 2 or 3.

28. The compound of claim 27, wherein R 1 is optionally replaced by one, two or three R 8 Substituted 5-10 membered heteroaryl.

29. The compound of claim 27 or 28, wherein R 1 is thiazolyl or pyridinyl, both of which are optionally substituted by one, two or three R 8 replace.

30. The compound of any one of claims 27-29, wherein R 1 Selected from the group consisting of: and ; Both of which are optionally replaced by one, two or three R 8 replace.

31. The compound of any one of claims 27-30, wherein the compound of formula II is a compound of formula IIa: or a pharmaceutically acceptable salt thereof.

32. The compound of any one of claims 27-31, wherein A' is N.

33. The compound of any one of claims 27-31, wherein A' is CH.

34. A compound as described in any one of claims 27-33, wherein R 2 It's a halogen.

35. A compound as described in any one of claims 27-34, wherein R 3 Is one or two R 5 Substituted 6-10 membered aryl.

36. A compound as described in any one of claims 27-35, wherein R 5 is optionally replaced by R 7 4-7 membered heterocyclyl substituted once, twice or three times.

37. A compound as described in any one of claims 27-36, wherein R 3 yes or .

38. A compound as described in any one of claims 27-37, wherein R 7 It is a C1-C6 alkyl group.

39. The compound of any one of claims 27-38, wherein n is 0 or 1.

40. The compound of claim 27, wherein A and A' are each independently CH or N; R 1 is optionally replaced by one, two or three R 8 substituted 5-10 membered heteroaryl; R 2 It is a halogen; Alternatively, two R 2 Together with the atoms to which they are attached, they form a 3-10 membered cycloalkyl or a 3-10 membered heterocycloalkyl; R 3 Is one or two R 5 substituted 6-10 membered aryl; R 5 is optionally replaced by R 7 a 4-7 membered heterocyclic group substituted once, twice or three times; R 7 is a C1-C6 alkyl group; R 8 is independently selected at each occurrence from C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C1-C3 alkylamine, halogen, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, (CH2) 1-4 OH, S(O) 0-2 H, S(O) 0- 2NH2 and CN group; and n is 0 or 1.

41. A compound as described in any one of claims 27-40, wherein the compound of formula II is selected from the following compounds or a group consisting of pharmaceutically acceptable salts thereof.

42. A pharmaceutical composition comprising the compound of any one of claims 1 to 41 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier.

43. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-41 or a pharmaceutical composition according to claim 42.

44. The method of claim 43, wherein the cancer is selected from the group consisting of lung cancer, colon cancer, breast cancer, endometrial cancer, thyroid cancer, glioma, squamous cell carcinoma, and prostate cancer.

45. The method of claim 43, wherein the cancer is non-small cell lung cancer (NSCLC).

46. ​​The method of any one of claims 43-45, wherein the method further comprises administering a second active agent.

47. A method of inhibiting a kinase in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-41 or a pharmaceutical composition according to claim 42.

48. The method of claim 47, wherein the kinase is EGFR.

49. A method of treating a kinase-mediated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-41 or a pharmaceutical composition according to claim 42.

50. The method of claim 49, wherein the kinase-mediated disorder is an EGFR-mediated disorder.

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