A substituted 2-arylaminopyrimidine compound, pharmaceutical composition and use thereof

Abstract

The present invention discloses a substituted 2-arylaminopyrimidine compound, a pharmaceutical composition, and its use, belonging to the field of chemical medicine. The present invention provides a novel 2-arylaminopyrimidine compound having a structure represented by general formula (I). This compound has enhanced metabolic stability and a longer metabolic half-life, exhibits higher inhibitory activity against activated or drug-resistant mutant forms of EGFR than wild-type EGFR, and can effectively reduce side effects. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of chemical medicine and relates to a substituted 2-arylaminopyrimidine compound, a pharmaceutical composition and use thereof. Background Art

[0002] The epidermal growth factor receptor (EGFR) belongs to the receptor tyrosine kinase family and, together with HER2, HER3, and HER4, forms the ErbB receptor family. When it binds to ligands such as epidermal growth factor (EGF), the receptor is activated and forms a dimer, further activating the phosphorylation of key tyrosine residues within the cell, ultimately activating downstream signaling pathways within the cell, such as the PI3K-AKT-mTOR pathway involved in cell apoptosis and the RAS-RAF-MEK pathway involved in cell cycle progression and proliferation. When genetic mutations occur in the EGFR domain, downstream signaling pathways are overexpressed, ultimately leading to the formation, proliferation, invasion, and migration of tumor cells, such as non-small cell lung cancer (NSCLC). Therefore, EGFR is one of the main targets for anticancer drug development.

[0003] Osimertinib (AZD9291) is a third-generation EGFR-TKI targeted drug. Although it has a high response rate against resistance caused by the L858R / T790M mutation, patients can also develop resistance (Clinical Cancer Research

[2015] , 17:3924-3933). In 2015, the first report (Nature Medicine

[2015] , 21:560-562) on the resistance analysis of AZD9291 in 15 patients was reported, in which a third mutation, namely EGFR C797S Mutation is one of the main mechanisms leading to drug resistance to AZD9291, accounting for about 40%. At the same time, resistance to AZD9291 was also reported in various conferences. In 2015 WCLC, Oxnard GR reported a resistance analysis of 67 patients, of which C797S accounted for ~22%; in 2017 ASCO, Piotrowska also reported 23 cases, of which C797S also accounted for about 22%; in 2022 WCLC, Winship Cancer Institute, Guardant Health and Blueprint Medicines jointly reported a resistance analysis of 65,273 adult patients with advanced NSCLC after 5 years of follow-up, and found that C797X has surpassed MET amplification to become the most common resistance mutation in patients receiving AZD9291 treatment. Among patients who may experience disease progression after AZD9291 as a first-line drug, EGFR C797X The incidence of mutation is 12.5%, which is 1.25 times that of MET amplification. C797XThe incidence of mutation is 2.4 times that of MET amplification. Except for the first year of AZD9291 first-line treatment, the incidence of MET amplification exceeds that of EGFR C797X ; In the next 2 to 5 years, EGFR C797X Both exceeded MET amplification. C797S mutation is the most common mutation type of C797X. Therefore, targeting C797S mutation to overcome AZD9291 resistance provides patients with a safer and more effective EGFR L858R / T790M / C797S Inhibitors have important research significance.

[0004]

[0005] In 2016, a compound EAI045 was reported that could overcome AZD9291 resistance by targeting C797S mutation (Nature

[2016] , 534:129-132). EAI045 is an allosteric inhibitor. After combining with EGFR monoclonal antibody, it showed good tumor suppression effect in the in vivo pharmacodynamic model of mice with L858R / T790M / C797S mutation; however, the compound failed to enter clinical research. In 2017, it was reported that the combination of brigatinib (AP26113) and EGFR monoclonal antibody could overcome AZD9291 resistance caused by C797S mutation (Nature Communications

[2017] , 8:14768). In PC9 (EGFR del19 / T790M / C797S ) The results of the mouse efficacy model showed that the combination of AP26113 and panitumumab or cetuximab showed good anti-tumor efficacy.

[0006]

[0007] Some current reports of 2-phenylaminopyrimidine inhibitors (e.g., WO2012051587A1, CN113354685A, CN113166110A) have shown therapeutic potential or characteristics for cancers that have metastasized to the CNS, but further efforts are needed to improve brain barrier permeability, enhance metabolic stability, and improve pharmacokinetic properties and druggability. Summary of the Invention

[0008] Problems to be solved by the invention

[0009] In order to solve the above-mentioned problems existing in the prior art, the present invention provides a new substituted 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof, which has enhanced metabolic stability and a longer metabolic half-life, shows higher inhibitory activity against activated or drug-resistant mutant forms of EGFR than wild-type EGFR, and can effectively reduce side effects.

[0010] The present invention also provides a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt thereof.

[0011] In addition, the present invention provides uses of the above-mentioned compound or a pharmaceutically acceptable salt thereof.

[0012] Solutions for solving problems

[0013] The present invention first provides a compound having a structure represented by general formula (I) or a pharmaceutically acceptable salt thereof,

[0014]

[0015] in:

[0016] X is selected from C and N;

[0017] R 1 Selected from H, C 1-6 Alkyl and C 3-6 Cycloalkyl; wherein the C 1-6 Alkyl and C 3-6 Cycloalkyl is optionally substituted with 0, 1, 2 or 3 R;

[0018] R 2 Selected from H, halogen, CN, OH, NO2, NH2, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-14 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, phenyl and 3-14 membered heterocyclic group; wherein said NH2, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-14 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, phenyl and 3-14 membered heterocyclyl are optionally substituted with 0, 1, 2 or 3 R's;

[0019] R 3 Selected from H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl and 5-6 membered heterocyclic group containing 1, 2 or 3 N or O atoms; wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl and 5-6 membered heterocyclyl containing 1, 2 or 3 N or O atoms are optionally substituted by 0, 1, 2 or 3 R";

[0020] R4 Selected from H, halogen, CN, NH2, C 1-4 Alkyl, C 1-4 Heteroalkyl, C 3-6 Cycloalkyl, phenyl and 5-6 membered heterocyclic group; wherein said NH2, C 1-4 Alkyl, C 1-4 Heteroalkyl, C 3-6 Cycloalkyl, phenyl and 5- to 6-membered heterocyclyl are optionally substituted with 0, 1, 2 or 3 R'';

[0021] R 5 and R 6 are independently selected from H, C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy; wherein the C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy is optionally substituted with 0, 1, 2 or 3 R"";

[0022] R, R', R", R'', R"" are independently selected from H, halogen, CN, OH, NH2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, C 1-6 Heteroalkyl, 3-6 membered heterocyclic group, phenyl group and 5-6 membered heteroaryl group;

[0023] "Hetero" means a heteroatom or heteroatom group, the C 1-6 Heteroalkyl, 3-14 membered heterocyclic group, 5-6 membered heterocyclic group, C 1-4 The "hetero" of heteroalkyl, 5-6 membered heterocyclic, 3-6 membered heterocyclyl, and 5-6 membered heteroaryl is independently selected from -C(=O)N(R a )-、-N(R b )-、-S(=O)2N(R c )-, -NH-, -O-, -S-, -C(=O)O-, -C(=O)-, -S(=O)2- and -N(R d )C(=O)N(R e )-; In any of the above cases, the number of heteroatoms or heteroatoms is independently selected from 1, 2 or 3, R a 、R b 、R c 、R d 、R eare independently selected from H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl.

[0024] In one embodiment of the present invention, R, R', R", R'', R"" are independently selected from H, F, Cl, Br, I, -CN, -OH, -NH2, -N(CH3)2, -CH3, CH3CH2-, -CH3CH2CH2, -CH(CH3)2, CH3O-, and

[0025] In one embodiment of the present invention, R 1 Selected from H, C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 0, 1, 2 or 3 R groups.

[0026] In one embodiment of the present invention, R 2 Selected from H, halogen, CN, OH, NO2, NH2, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclic groups; wherein the NH2, C 3-12 Cycloalkyl and 3-12 membered heterocyclyl are optionally substituted with 0, 1, 2 or 3 R's.

[0027] In one embodiment of the present invention, R 2 Preferred are piperidine rings, piperazine rings, and morpholine rings substituted with 0, 1, 2, or 3 R's.

[0028] In one embodiment of the present invention, R 3 Selected from H, F, Cl, Br, -CH3, CH3CH2-, (CH3)2CH-.

[0029] In one embodiment of the present invention, R 4 Select H, F, Cl, Br, I, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, phenyl, Among them, the CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, phenyl, Optionally substituted with 0, 1, 2 or 3 R''.

[0030] In one embodiment of the present invention, R 5 and R 6 are independently selected from H, C 1-6 Alkyl, C1-6 Alkoxy and C 3-6 Cycloalkyl, wherein the C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl is optionally substituted with 1, 2 or 3 R"".

[0031] In one embodiment of the present invention, R 5 and R 6 are independently selected from H, C 2-3 Alkyl; C 2-3 Alkyl is optionally substituted with 0, 1, 2 or 3 R"".

[0032] In one embodiment of the present invention, the compound is specifically selected from:

[0033]

[0034]

[0035] In one embodiment of the present invention, the pharmaceutically acceptable salt is an inorganic salt or an organic salt. The inorganic salt includes hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, succinate, glutarate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, p-toluenesulfonate, and ascorbate.

[0036] In one embodiment of the present invention, the pharmaceutically acceptable salt is selected from hydrochloride, sulfate, succinate or methanesulfonate.

[0037] The present invention also provides a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, excipient or diluent.

[0038] The present invention also provides use of the compound or a pharmaceutically acceptable salt thereof in preparing a drug for treating diseases, especially cancers, mediated by EGFR activation or drug-resistant mutants in mammals, especially humans.

[0039] The present invention also provides the use of the compound or a pharmaceutically acceptable salt thereof, wherein the cancer is non-small cell lung cancer.

[0040] The present invention also provides use of the compound or a pharmaceutically acceptable salt thereof in combination with an antitumor agent in the preparation of a drug for a disease mediated by an EGFR activated or drug-resistant mutant, wherein the antitumor agent is selected from the following:

[0041] (i) Anti-tumor drugs that act on DNA structure;

[0042] (ii) antitumor drugs that affect nucleic acid synthesis;

[0043] (iii) anti-tumor drugs that affect nucleic acid transcription;

[0044] (iv) anti-tumor drugs that target tubulin synthesis;

[0045] (v) cell signaling pathway inhibitors such as epidermal growth factor receptor inhibitors;

[0046] (vi) Anti-tumor monoclonal antibodies.

[0047] Effects of the Invention

[0048] The present invention provides a new 2-arylaminopyrimidine inhibitor of an activated mutant form of the epidermal growth factor receptor, which has better pharmacodynamics and higher metabolic stability, shows higher inhibitory activity against activated or drug-resistant mutant forms of EGFR than wild-type EGFR, and can effectively reduce side effects such as rash and diarrhea. DETAILED DESCRIPTION

[0049] The technical solution of the present invention will be described in detail below with reference to embodiments.

[0050] In the present invention, C 1-4 Alkyl, C 1-6 Alkyl, C 1-14 Alkyl refers to a straight or branched saturated hydrocarbon group having 1 to 4 carbon atoms, 1 to 6 carbon atoms, or 1 to 14 carbon atoms, respectively, and examples thereof include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, and tert-butyl;

[0051] C 2-6 Alkenyl refers to a straight or branched chain hydrocarbon group having one or more double bonds and 2 to 6 carbon atoms;

[0052] C 2-6 Alkynyl refers to a straight or branched chain hydrocarbon group having one or more triple bonds and 2 to 6 carbon atoms;

[0053] C 3-6 Cycloalkyl, C 3-14 Cycloalkyl refers to a saturated monocyclic or polycyclic hydrocarbon group having 3 to 6 ring carbon atoms or 3 to 14 ring carbon atoms, respectively. Examples thereof include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0054] C 3-6Cycloalkenyl refers to a monocyclic or polycyclic hydrocarbon group having one or more double bonds and 3 to 6 ring carbon atoms;

[0055] C 4-6 Cycloalkynyl refers to a monocyclic or polycyclic hydrocarbon group having one or more triple bonds and 4 to 6 ring carbon atoms;

[0056] C 1-4 Heteroalkyl, C 1-6 Heteroalkyl refers to a linear or branched monovalent saturated alkyl group having 1 to 4 carbon atoms, 1 to 6 carbon atoms, respectively, in which 1 to 3 hydrogen atoms are replaced by heteroatoms or heteroatom groups;

[0057] 3- to 6-membered heterocyclic group, 5- to 6-membered heterocyclic group, and 3- to 14-membered heterocyclic group refer to monocyclic or polycyclic hydrocarbon groups having 3 to 6 ring carbon atoms, 5 to 6 ring carbon atoms, and 3 to 14 ring carbon atoms, respectively, in which 1 to 3 hydrogen atoms are replaced by heteroatoms or heteroatom groups;

[0058] 5- to 6-membered heteroaryl refers to a monocyclic or polycyclic aromatic group having 5 to 6 ring carbon atoms, wherein 1 to 3 hydrogen atoms are replaced by heteroatoms or heteroatom groups;

[0059] C 1-14 Alkoxy refers to a straight or branched saturated hydrocarbon group having 1 to 14 carbon atoms connected by an oxygen bridge, examples of which include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy;

[0060] C 3-6 Cycloalkyloxy refers to a monocyclic or polycyclic hydrocarbon group having 3 to 6 ring carbon atoms linked by an oxygen bridge;

[0061] Indicates the point from which a substituent is attached.

[0062] As used herein, the term "disease" refers to any condition or disorder that damages or interferes with the normal function of a cell, organ, or tissue.

[0063] As used herein, the term "inhibitor" refers to a compound or agent that has the ability to inhibit the biological function of a targeted protein or polypeptide, for example, by inhibiting the activity or expression of the protein or polypeptide.

[0064] As used herein, the term "anti-neoplastic agent" refers to any agent useful in the treatment of neoplastic disorders.

[0065] As used herein, the term "pharmaceutically acceptable" refers to compositions that are suitable for use in contact with the tissues of humans and other mammals without excessive toxicity, irritation, allergic response, etc., within a reasonable medical range and at a reasonable benefit / risk ratio. "Pharmaceutically acceptable salt" refers to any non-toxic salt that, upon administration to a recipient, is capable of providing, directly or indirectly, a compound or prodrug of the compound of the present invention.

[0066] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or pharmaceutical composition described herein that is sufficient to achieve the intended application, including, but not limited to, treating a disease. In some embodiments, the amount is measured to be effective for killing or inhibiting the growth or spread of cancer cells; the size or number of tumors; or the severity level, stage, and progression of the cancer. The therapeutically effective amount can vary depending on the intended application, such as in vitro or in vivo, the condition and severity of the disease, the age, weight, or mode of administration of the subject, etc. The term also applies to a dose that will induce a specific response in the target cells, such as a reduction in cell migration. The specific dose will depend on, for example, the specific compound selected, the species of the subject and their age / existing health condition or risk of health condition, the route of administration, the severity of the disease, administration in combination with other agents, the timing of administration, the tissue to which it is administered, and the drug delivery device, etc.

[0067] "Administration" or "administering" a subject compound in the context of the present invention means providing the subject in need of treatment with a compound of the present invention.

[0068] The compounds of the present invention may contain one or more asymmetric centers and therefore occur as racemates and racemic mixtures, single enantiomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present invention. The compounds of the present invention may also exhibit multiple tautomeric forms, in which case the present invention expressly includes all tautomeric forms of the compounds described herein. All such isomeric forms of such compounds are included in the present invention. All crystalline forms of the compounds described herein are expressly included in the present invention.

[0069] <Compound or Pharmaceutically Acceptable Salt thereof>

[0070] The present invention provides a novel 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof in the form of an activated mutation of epidermal growth factor receptor, the structural formula of which is shown in general formula (I):

[0071]

[0072] in:

[0073] X is selected from C and N;

[0074] R1 Selected from H, C 1-6 Alkyl and C 3-6 Cycloalkyl, wherein the C 1-6 Alkyl and C 3-6 Cycloalkyl is optionally substituted with 0, 1, 2 or 3 R;

[0075] R 2 Selected from H, halogen, CN, OH, NO2, NH2, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-14 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, phenyl and 3-14 membered heterocyclic group, wherein the NH2, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-14 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, phenyl and 3-14 membered heterocyclyl are optionally substituted with 0, 1, 2 or 3 R's;

[0076] R 3 Selected from H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl and 5-6 membered heterocyclic group containing 1, 2 or 3 N or O atoms, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl and 5-6 membered heterocyclyl containing 1, 2 or 3 N or O atoms are optionally substituted by 0, 1, 2 or 3 R";

[0077] R 4 Selected from H, halogen, CN, NH2, C 1-4 Alkyl, C 1-4 Heteroalkyl, C 3-6 Cycloalkyl, phenyl and 5-6 membered heterocyclic group, wherein the NH2, C 1-4 Alkyl, C 1-4 Heteroalkyl, C 3-6 Cycloalkyl, phenyl and 5- to 6-membered heterocyclyl are optionally substituted with 0, 1, 2 or 3 R'';

[0078] R 5 and R 6 are independently selected from H, C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6Cycloalkyloxy, wherein the C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy is optionally substituted with 0, 1, 2 or 3 R"";

[0079] R, R', R", R'', R"" are independently selected from H, halogen, CN, OH, NH2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, C 1-6 Heteroalkyl, 3-6 membered heterocyclic group, phenyl group and 5-6 membered heteroaryl group;

[0080] "Hetero" means a heteroatom or heteroatom group, the C 1-6 Heteroalkyl, 3-14 membered heterocyclic group, 5-6 membered heterocyclic group, C 1-4 The “hetero” of heteroalkyl, 5- to 6-membered heterocyclic, 3- to 6-membered heterocyclyl, and 5- to 6-membered heteroaryl is independently selected from —C(═O)N(Ra)—, —N(Rb)—, —S(═O)2N(Rc)—, —NH—, —O—, —S—, —C(═O)O—, —C(═O)—, —S(═O)2-, and —N(Rd)C(═O)N(Re)—; in any of the above cases, the number of heteroatoms or heteroatom groups is independently selected from 1, 2, or 3, and R a 、R b 、R c 、R d 、R e are independently selected from H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl.

[0081] In certain embodiments of the present invention, X is selected from C and N; further, X is preferably C.

[0082] In certain embodiments of the present invention, R, R', R", R'', R"" are independently selected from H, halogen, CN, OH, NH2, -N(CH3)2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, C1-6 heteroalkyl, 3- to 6-membered heterocyclic group, phenyl and 5- to 6-membered heteroaryl.

[0083] In certain embodiments of the present invention, R 1 Selected from H, C 1-6 Alkyl and C 3-6 Cycloalkyl, wherein the C 1-6 Alkyl and C 3-6 The cycloalkyl group is optionally substituted with 0, 1, 2 or 3 R; further, R 1 Selected from C 1-6 Alkyl, R is selected from H or OH; further R 1 R is selected from methyl, ethyl or 2-propyl, and R is selected from H.

[0084] In certain embodiments of the present invention, R 2 Selected from H, halogen, CN, OH, NO2, NH2, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-14 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, phenyl and 3-14 membered heterocyclic group, wherein the NH2, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-14 Cycloalkyl, C 3-6 Cycloalkenyl, C 4-6 Cycloalkynyl, phenyl and 3-14 membered heterocyclic groups are optionally substituted by 0, 1, 2 or 3 R's; further, R 2 Selected from C 1-6 heteroalkyl or 3-14 membered heterocyclic group, R' is selected from H or OH; further R 2 is selected from morpholine, 1-isopropylpiperazine, 4-dimethylaminopiperidine or N-methylpiperazine, and R' is selected from H.

[0085] In certain embodiments of the present invention, R 3 Selected from H, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl and 5-6 membered heterocyclic group containing 1, 2 or 3 N or O atoms, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl and 5-6 membered heterocyclic groups containing 1, 2 or 3 N or O atoms are optionally substituted by 0, 1, 2 or 3 R"; further, R 3 Selected from C 1-6Alkyl, R" is selected from H or OH; further R 3 is selected from methyl, and R" is selected from H.

[0086] In certain embodiments of the present invention, R 4 Selected from H, halogen, CN, NH2, C 1-4 Alkyl, C 1-4 Heteroalkyl, C 3-6 Cycloalkyl, phenyl and 5-6 membered heterocyclic group, wherein the NH2, C 1-4 Alkyl, C 1-4 Heteroalkyl, C 3-6 Cycloalkyl, phenyl and 5-6 membered heterocyclic groups are optionally substituted by 0, 1, 2 or 3 R''s; further, R 4 is selected from H, and R'' is selected from H.

[0087] In certain embodiments of the present invention, R 5 and R 6 are independently selected from H, C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy, wherein the C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy is optionally substituted with 0, 1, 2 or 3 R""; further, R 5 Preferably H or C 1-14 Alkyl, R 6 Preferably C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl or C 3-6 Cycloalkyloxy, R"" is selected from H or OH; further, R 5 Selected from methyl, R 6 is selected from methyl or 1-propyl, R"" is selected from H; or, R 5 Selected from H, R 6 is selected from 1-propyl, 2-methoxyethyl or 4-methoxycyclohexyl, R"" is selected from H; in addition, R 5 Selected from H, R 6 is selected from ethyl, 1-propyl, 1-butyl, 2-propyl, 2-(3-methyl)butyl, (S)-2-propyl, (R)-2-propyl or 4-cyclohexyl, and R"" is selected from OH.

[0088] The compound of general formula (I) includes its pharmaceutically acceptable salt. The pharmaceutically acceptable salt of the present invention is an inorganic salt or an organic salt. The inorganic salt includes hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, succinate, glutarate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, p-toluenesulfonate, and ascorbate. Preferably, from the perspective of drugability, the salt of the present invention is hydrochloride, sulfate, succinate, or methanesulfonate.

[0089] It should be understood that certain compounds of formula (I) or pharmaceutically acceptable salts thereof may exist in solvate forms as well as unsolvate forms, such as, for example, water and HCl. It should be understood that the present invention encompasses all such solvate forms possessing activating mutant EGFR inhibitory activity.

[0090] The synthesis of the compounds of the general formula (I) of the present invention can be achieved by ordinary synthetic chemists. The references mentioned in the background art are incorporated herein by reference in their entirety. The preparation methods are described in detail in the examples.

[0091] <Pharmaceutical Composition>

[0092] The present invention provides a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, excipient or diluent.

[0093] The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into solid preparations for oral administration, including, but not limited to, capsules, tablets, pills, powders, granules, etc. In these solid dosage forms, the compound of the general formula (I) of the present invention is mixed as an active ingredient with at least one conventional inert excipient (or carrier), for example, sodium citrate or dicalcium phosphate. Or mixed with the following ingredients: (1) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (2) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.; (3) humectants, such as glycerol, etc.; (4) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, etc.; (5) solubilizers, such as paraffin, etc.; (6) absorption accelerators, such as quaternary ammonium compounds, etc.; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate, etc.; (8) adsorbents, such as kaolin, etc.; (9) lubricants, such as talc, calcium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Capsules, tablets and pills may also contain buffers.

[0094] Solid dosage forms such as tablets, sugar pills, capsules, pills, and granules can be coated or microencapsulated with coatings and shell materials such as enteric coatings and other materials known in the art. They can contain opacifying agents, and the release of the active ingredient in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If desired, the active ingredient can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0095] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, and the like. In addition to the compound of formula (I) or its pharmaceutically acceptable salt as the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, or mixtures thereof. In addition to these inert diluents, the liquid dosage form of the present invention may also include conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0096] The suspending agents include, for example, ethoxylated stearyl alcohol, polyoxyethylene sorbitol, and sorbitan, microcrystalline cellulose, agar, etc., or a mixture of these substances.

[0097] The compounds of the present invention and their pharmaceutically acceptable salts can be formulated into dosage forms for parenteral injection, including, but not limited to, physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions and dispersions. Suitable carriers, diluents, solvents, and excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0098] The compound of the present invention or its pharmaceutically acceptable salt can be formulated into dosage forms for topical administration, including ointments, powders, suppositories, drops, sprays and inhalants, etc. The compound of the present invention of general formula (I) or its pharmaceutically acceptable salt as the active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and optional preservatives, buffers, and propellants as needed.

[0099] The compound of formula (I) of the present invention or its pharmaceutically acceptable salt will be given to mammals with a unit dose in the range of 0.01-2000 mg / kg, particularly 2.5-1000 mg / kg, particularly 5-500 mg / kg, and this should provide an effective dose. However, the daily dose will necessarily vary depending on the severity of the treated host, the specific route of administration, and the disease being treated. Therefore, the optimal dose can be determined by the practitioner treating any particular patient.

[0100] <Purpose>

[0101] The present invention provides a compound of formula (I) as defined above and its pharmaceutically acceptable salts for use in preparing drugs for treating diseases mediated by EGFR activating or drug-resistant mutants in mammals, especially humans, especially cancer.

[0102] In the present invention, the EGFR in the form of an activated mutant or a drug-resistant mutant can be, for example, an L858R activating mutant, an Exon 19 deletion activating mutant, a T790M resistance mutant, and / or a C797S resistance mutant. Therefore, the disease, disorder, condition, or condition mediated by an EGFR activating or drug-resistant mutant can be, for example, a disease, disorder, condition, or condition mediated by an L858R activating mutant, an Exon 19 deletion activating mutant, a T790M resistance mutant, and / or a C797S resistance mutant. The present invention is particularly applicable to diseases, disorders, conditions, or conditions mediated by EGFR drug-resistant mutants, such as T790M resistance mutants and C797S resistance mutants. Cancer types that may be susceptible to treatment with a compound of formula (I) or a pharmaceutically acceptable salt thereof include, but are not limited to, ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, malignant glioma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple myeloma, melanoma, and mesothelioma. Preferably, the cancer includes non-small cell lung cancer and metastatic non-small cell lung cancer.

[0103] In the treatment of cancer according to the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to a mammal, more particularly a human.

[0104] The active treatment of the EGFR in the form of activated mutants and the EGFR in the form of drug-resistant mutants described in the present invention can be applied as a single therapy or, in addition to the compounds of the present invention, can involve conventional surgery or radiotherapy (such as the WBRT described in the present invention), can be administered in combination with other pharmaceutically acceptable therapeutic agents, and can be combined with other anti-tumor drugs. This combined treatment can be achieved by using the components of the treatment simultaneously, sequentially or separately. The therapeutic tumor agents include but are not limited to: anti-tumor drugs that act on the chemical structure of DNA, such as cisplatin, anti-tumor drugs that affect nucleotide synthesis, such as methotrexate, 5-fluorouracil, etc., anti-tumor drugs that affect nucleic acid transcription, such as doxorubicin, epirubicin, aclarubicin, etc., anti-tumor drugs that act on microtubule synthesis, such as paclitaxel, vinorelbine, etc., aromatase inhibitors such as aminoglutethimide, letrozole, and arimide, etc., cell signaling pathway inhibitors such as epidermal growth factor receptor inhibitors imatinib, Gefitinib, erlotinib, afatinib, osimertinib, etc., 6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzimidazole-5-carboxylic acid (2-hydroxy-ethoxy)-amide or a pharmaceutically acceptable salt thereof, 1-[(1S)-1-(imidazo[1,2-a]pyridin-6-yl)ethyl]-6-(1-methyl-1H-pyrazol-4-yl)-1H[1,2,3]triazolo[4,5-b]pyrazine or a pharmaceutically acceptable salt thereof. Anti-tumor monoclonal antibodies, such as anti-CTLA-4 antibodies, immunosuppressants PD-1, PD-L1, OX40 agonist antibodies, etc., the components to be combined can be administered simultaneously or sequentially, in the form of a single preparation or in the form of different preparations. The combination includes not only a combination of one or other active agents of the compound of the present invention, but also a combination of two or more other active agents of the compound of the present invention.

[0105] The following examples illustrate, but are not intended to limit, the methods for synthesizing compounds of formula (I). All temperatures are in degrees Celsius. Unless otherwise stated, all evaporations were performed under reduced pressure. Unless otherwise stated, reagents were purchased from commercial suppliers and used without further purification. The structures of final products, intermediates, and starting materials were confirmed by standard analytical methods, such as elemental analysis and spectral characterization, e.g., MS and NMR. Abbreviations used are those conventional in the art.

[0106] Intermediate A

[0107] 5-Fluoro-4-methyl-2-nitrophenol

[0108]

[0109] 3-Fluoro-4-methylphenol (10.00 g, 79.31 mmol) and benzyltriethylammonium chloride (1.77 g, 7.93 mmol) were dissolved in dichloromethane (80 mL). 65% concentrated nitric acid (7.35 g, 79.31 mmol) was slowly added dropwise at 0°C. Stirring was continued for 0.5 h. After the reaction, saturated sodium bicarbonate solution was added to the reaction mixture under an ice bath to adjust the pH to 8.0-9.0. Extraction was then performed. The aqueous phase was further extracted with dichloromethane (40 mL x 2). The combined organic phases were washed sequentially with water (40 mL) and saturated sodium chloride (40 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. Purification by column chromatography [PE:EA = 200:1 (v / v)] afforded Intermediate A (11.30 g) as a reddish-brown oil in a yield of 83.2%. 1 H-NMR (400MHz, CDCl3-d) δ: 10.63 (s, 1H), 7.99 (d, J = 7.8Hz, 1H), 6.80 (d, J = 10.1Hz, 1H), 2.26 (d, J = 1.9Hz, 3H). ESI-HRMS(m / z):170.1257[M+H] - .

[0110] Intermediate B

[0111] 1-Fluoro-5-methoxy-2-methyl-4-nitrobenzene

[0112]

[0113] Intermediate A (11.30 g, 66.00 mmol) was dissolved in N,N-dimethylformamide (100 mL), and anhydrous potassium carbonate (13.71 g, 99.00 mmol) and iodomethane (14.12 g, 99.30 mmol) were added. The mixture was stirred at room temperature overnight. After the reaction, ice water (220 mL) was added to precipitate a solid. The mixture was stirred at room temperature for 1 hour, allowed to stand for 20 minutes, and filtered. The filter cake was washed with water (50 mL) and dried in a vacuum oven at 50°C overnight to obtain an off-white solid, Intermediate B (11.61 g), with a yield of 94.6%. The solid was used directly in the next reaction without further treatment. 1 H-NMR (400MHz, DMSO-d6) δ: 7.94 (d, J = 8.0 Hz, 1H), 7.27 (d, J = 11.7 Hz, 1H), 3.91 (s, 3H), 2.21 (d, J = 2.0 Hz, 3H). ESI-HRMS(m / z):186.0627[M+H] + .

[0114] Intermediate C

[0115] 1-(1-(5-methoxy-2-methyl-4-nitrophenyl)piperidin-4-yl)-4-methylpiperazine

[0116]

[0117] Intermediate B (11.61 g, 62.52 mmol) was dissolved in N,N-dimethylformamide (200 mL), and anhydrous potassium carbonate (17.33 g, 125.04 mmol) and 1-methyl-4-(piperidin-4-yl)piperazine (17.24 g, 93.73 mmol) were added. The mixture was stirred at 120 ° C for 4 h. After cooling to room temperature after the reaction, the reaction solution was slowly added to ice water (400 mL). Solid precipitated, stirred at room temperature for 1.5 h, and then allowed to stand for 30 min. The filter cake was washed with water (60 mL) and dried in a vacuum drying oven at 50 ° C overnight to obtain yellow solid intermediate C (20.02 g) with a yield of 92.1%. It can be directly used in the next reaction without post-treatment. 1 H-NMR (400MHz, CDCl3-d) δ: 7.81 (s, 1H), 6.54 (s, 1H), 3.93 (s, 3H), 3.34 (d, J = 11.9Hz, 2H), 2.96 (s,4H),2.88(s,4H),2.73~2.59(m,5H),2.30(s,3H),2.23(s,3H),1.71(qd,J=12.0,3.8Hz,2H). ESI-HRMS(m / z):349.2045[M+H] + .

[0118] Intermediate D

[0119] 2-Methoxy-5-methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline

[0120]

[0121] Intermediate C (20.02 g, 57.51 mmol) was added to a mixed solution of ethanol and water [210 mL, EtOH:H2O=2:1 (V / V)], and ammonium chloride (21.52 g, 403.07 mmol) was added. The mixture was refluxed and stirred for 1 h to fully dissolve it. Iron powder (16.13 g, 288.05 mmol) was then added, and the reaction was continued at 80°C for 4 h. After the reaction was completed, the reaction solution was filtered through diatomaceous earth to remove the iron powder while it was hot. The filter cake was washed with ethanol until no fluorescence was detected by TLC. The filtrate was collected and concentrated under reduced pressure. The product was dried in a vacuum drying oven at 50°C overnight to obtain a black solid intermediate D (17.94 g) with a yield of 97.8%. The product was directly used in the next reaction without post-treatment. 1H-NMR (400MHz, CD3OD-d4) δ: 6.62 (s, 1H), 6.60 (s, 1H), 3.80 (s, 3H), 3.05 (d, J = 11.6Hz, 2H), 2.81~2.44 (m, 10 H), 2.36 (dt, J = 11.5, 3.8Hz, 1H), 2.31 (s, 3H), 2.14 (s, 3H), 2.00 ~ 1.92 (m, 2H), 1.66 (qd, J = 12.0, 3.9Hz, 2H). ESI-HRMS(m / z):319.2355[M+H] + .

[0122] Intermediate E

[0123] N-(2-Methoxy-5-methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)formamide

[0124]

[0125] Acetic anhydride (1.51 g, 14.70 mmol) and formic acid (0.76 g, 16.51 mmol) were thoroughly mixed and stirred at reflux for 4 h. The reaction was then cooled to room temperature and added to Intermediate D (1.50 g, 4.71 mmol) dissolved in tetrahydrofuran (15 mL). The mixture was stirred at room temperature for 2 h. After the reaction, saturated sodium bicarbonate solution was added to adjust the pH to 8.0-9.0. The mixture was extracted with dichloromethane (25 mL x 4). The organic phases were combined, washed with saturated sodium chloride (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain Intermediate E (1.48 g) as a pale yellow solid in a yield of 90.5%. The product was used directly in the next step without post-treatment. ESI-HRMS (m / z): 347.2331 [M+H] + .

[0126] Intermediate F

[0127] 1-(2-(Methylthio)pyrimidin-4-yl)-3-phenyl-1H-pyrazole-4-carbaldehyde

[0128]

[0129] 2-Methylthio-4-chloropyrimidine (5.11 g, 32.02 mmol) was fully dissolved in N,N-dimethylformamide (35 mL). Anhydrous potassium carbonate (8.04 g, 58.22 mmol) and 3-phenyl-1H-pyrazole-4-carbaldehyde (L-2, 5.00 g, 29.13 mmol) were added and stirred at 50°C for 12 h. After the reaction was completed and cooled to room temperature, the reaction solution was slowly added to ice water (80 mL). After stirring for 3 h, a solid precipitated. After standing for 20 min, the resulting solid was filtered, the filter cake was washed with water (10 mL), and dried in a vacuum oven at 50°C overnight to obtain an off-white solid intermediate F (7.01 g) with a yield of 81.4%. The product was used directly in the next reaction without post-treatment. ESI-HRMS (m / z): 297.0735 [M+H] + .

[0130] Intermediate G

[0131] 1-(2-(Methylsulfonyl)pyrimidin-4-yl)-3-phenyl-1H-pyrazole-4-carbaldehyde

[0132]

[0133] Intermediate F (7.01 g, 23.72 mmol) and ammonium molybdate tetrahydrate (2.92 g, 2.37 mmol) were fully dissolved in ethanol (50 mL). 30% hydrogen peroxide solution (26.82 g, 236.53 mmol) was slowly added dropwise at room temperature. The mixture was stirred for 2 h. After the reaction, water (70 mL) was added and the mixture was extracted with dichloromethane (70 mL × 3). The organic phases were combined and washed with 10% sodium sulfite solution (30 mL × 2), and finally with water (70 mL) and saturated sodium chloride (60 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain Intermediate G (6.53 g) as a white solid in an 84.3% yield. The product was used directly in the next step without post-treatment. ESI-HRMS (m / z): 328.1768 [M+H] + .

[0134] Intermediate H

[0135] 1-(2-((2-methoxy-5-methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)-3-phenyl-1H-pyrazole-4-carbaldehyde

[0136]

[0137] Under nitrogen, add Intermediate E (1.48 g, 4.27 mmol) to ultra-dry tetrahydrofuran (9 mL), followed by ultra-dry N,N-dimethylacetamide (6 mL) to fully dissolve the mixture. The temperature is then lowered to 5°C, and a solution of ultra-dry sodium tert-butoxide (6 mL) dissolved in 1.0 mol / L tetrahydrofuran is slowly added dropwise. Stir for 2 h after the addition is complete. The reaction temperature was then adjusted to room temperature, and Intermediate G (1.68 g, 5.12 mmol) was added. Stirring was continued for 1 h. After the reaction, saturated sodium bicarbonate solution was added to the reaction solution to adjust the pH to 8.0-9.0. The mixture was extracted with dichloromethane (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride (40 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was separated and purified by column chromatography [DCM:MeOH = 15:1 (v / v)] to obtain Intermediate H (1.06 g) as a pale yellow solid in a yield of 43.9%. ESI-HRMS (m / z): 567.3224 [M+H]. + .

[0138] Example 1

[0139] 4-(4-((dimethylamino)methyl)-3-phenyl-1H-pyrazol-1-yl)-N-(2-methoxy-5-methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)pyrimidin-2-amine

[0140]

[0141] Intermediate H (1.00 g, 1.80 mmol) was added to methanol (20 mL), followed by dimethylamine (0.24 g, 5.31 mmol), glacial acetic acid (2 drops), and anhydrous magnesium sulfate (1.73 g, 14.12 mmol). The mixture was stirred at 70°C for 2 h. After the reaction, the temperature was adjusted to room temperature, sodium cyanoborohydride (3.31 g, 5.32 mmol) was added, and the mixture was stirred overnight. After the reaction, water (30 mL) was added to the reaction mixture to quench the reaction, followed by extraction with dichloromethane (30 mL x 6). All organic phases were combined, washed with saturated sodium chloride (60 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This was separated and purified by column chromatography [DCM:MeOH = 10:1 (v / v)] to obtain the desired product, Example 1 (0.21 g), in a yield of 19.5%. 1H NMR (400MHz, DMSO-d6) δ: 8.49 (s, 1H), 8.30 (s, 1H), 7.94 (d, J = 7.7Hz, 2H), 7.74 (s, 1H), 7.50 (t, J=7.5Hz,3H),7.44(d,J=7.2Hz,1H),7.25(d,J=5.4Hz,1H),6.73(s,1H),3.81(s,3H),3.42(s,2H ),3.11(d,J=11.1Hz,2H),2.62(dd,J=24.5,12.7Hz,8H),2.33(dd,J=6.9,3.6Hz,3H),2.23(s,6 H),2.21(s,6H),1.87(d,J=12.1Hz,2H),1.59(d,J=13.8Hz,2H).ESI-HRMS(m / z):596.3786[M+H] + .

[0142] Salt formation:

[0143] 4-(4-((dimethylamino)methyl)-3-phenyl-1H-pyrazol-1-yl)-N-(2-methoxy-5-methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)pyrimidin-2-amine hydrochloride

[0144]

[0145] Example 1 (0.21 g) was dissolved in acetonitrile (2 mL), and 1N HCl (2 mL) was slowly added with stirring. After stirring for a while, the solvent was removed by freeze-drying to obtain a yellow solid, which was the hydrochloride salt of Example 1 (0.26 g). 1 H-NMR(400MHz,D2O)δ:8.35(s,1H),8.21(s,1H),7.79(d,J=7.7Hz,2H),7.64(s,1H),7.42(t,J= 7.5Hz,3H),7.33(d,J=7.2Hz,1H),7.11(d,J=5.4Hz,1H),6.58(s,1H),3.59(s,3H),3.31(s,2H) ,3.03(d,J=11.1Hz,2H),2.52(dd,J=24.5,12.7Hz,8H),2.22(dd,J=6.9,3.6Hz,3H),2.12(s,6H ),2.10(s,6H),1.67(d,J=12.1Hz,2H),1.46(d,J=13.8Hz,2H).ESI-HRMS(m / z):596.3786[M+H] + .

[0146] Examples 2-20 (see Table 1) were synthesized by operations similar to those of Example 1 to obtain the desired products.

[0147] Table 1 Structure and mass spectrometry data of Examples 2-20

[0148]

[0149]

[0150]

[0151] Note: Preparation of Example 7: Using racemic DL-aminopropanol as the substrate, Intermediate H (1.00 g, 1.80 mmol) was added to methanol (20 mL), followed by DL-aminopropanol (0.39 g, 5.20 mmol), glacial acetic acid (2 drops), and anhydrous magnesium sulfate (1.73 g, 14.12 mmol). The mixture was stirred at 70°C for 2 h. After the reaction, the temperature was adjusted to room temperature, sodium cyanoborohydride (3.31 g, 5.32 mmol) was added, and the mixture was stirred overnight. After the reaction, water (30 mL) was added to the reaction solution to quench the reaction, and then dichloromethane (30 mL×6) was added for extraction. All organic phases were combined, washed with saturated sodium chloride (60 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was separated and purified by column chromatography [DCM:MeOH=10:1 (v / v)] to obtain the corresponding racemic product (0.15 g) in a yield of 13.54%.

[0152] The preparation process of Examples 9 and 10 refers to Example 7, and D-aminopropanol and L-aminopropanol substrates are respectively used to replace DL-aminopropanol to prepare the corresponding chiral products.

[0153] Example 20 Activity Test

[0154] Assay 1: Lance screening to determine EGFR inhibitory activity

[0155] IC was determined by Lance Ultra assay 50 The assay consists of two steps: an enzymatic reaction and a detection step. First, EGFR L858R / T790M / C797S A mixture of the test compound, various concentrations of the test compound, a peptide substrate, and adenosine triphosphate (ATP) is incubated in assay buffer. When EGFR is phosphorylated by the peptide substrate, phosphorylation can be detected using specific antibodies and TR-FRET assay technology.

[0156] Detailed description: The Lance screening assay was performed as a 384-well TR-FRET-based assay. In the first step, EGFR L858R / T790M / C797SIncubate for 15 minutes at 22°C in assay buffer containing different concentrations of test compound or without test compound (as negative control). The assay buffer contains 50mM HEPES pH 7.5, 10mM MgCl2, 0.01% BSA, 0.01% Tween-20 and 2mM dithiothreitol (DTT). Echo 555 (Labcyte) is used to distribute the compound solution. Then, in a second step, purified ULight-labeled peptide substrate and ATP are added and the reaction mixture is incubated at 22°C for 25 minutes. The pharmacologically relevant assay volume is 5μl. During the incubation period of the reaction mixture, the final concentration in the assay is 0.3nM EGFR L858R / T790M / C797S , 50 nM peptide substrate and 0.5 μM ATP. The enzymatic reaction is terminated by adding EDTA. Phosphorylation of EGFR mediated by the reaction in the presence of ATP can be detected using a specific antibody labeled with the fluorophore europium (Eu). For this purpose, 2 μl of a stop solution containing the antibody (12.5 mM HEPES pH 7.5, 125 mM EDTA, 30 mM sodium hydroxide, 300 mM potassium fluoride, 0.006% Tween-20, 0.21 nM anti-tyrosine phosphorylation-LANCE-Eu antibody (PT66), 15 nM ULight-peptide substrate) were added to the reaction mixture. After 2 hours of signal development, the plate was analyzed in an EnVision (PerkinElmer) microplate reader using TRF mode and laser excitation. After excitation of the donor europium at 340 nm, the fluorescence emitted by ULight at 665 nm and at 615 nm from the donor Eu was measured. The amount of phosphorylated peptide substrate is directly proportional to the ratio of the amount of light emitted at 665 nm and at 615 nm, i.e. the ratio of relative fluorescence units (rfu). The data were processed using Genedata Screener software. In particular, the IC was determined in a conventional manner by fitting a dose-response curve to the data points using nonlinear regression analysis. 50 value.

[0157] The structural formula of the reference compound (commercially available Lazertinib):

[0158] The experimental results are shown in Table 2.

[0159] Table 2 Data of activity determination of Examples of the present invention and reference compound (commercially available Lazertinib)

[0160]

[0161]

[0162] Assay 2: Evaluation of Compound Stability Using Human Liver Microsomes

[0163] The liver microsomal enzyme stability of the example compounds was compared with that of lazertinib.

[0164] Assay System: The metabolic stability of the compounds of this invention was tested using liver microsomes collected from male and female individuals using 1 mM NADPH. Samples were analyzed using a mass spectrometer. HRMS was used to determine the peak area response ratio (peak area corresponding to the test compound or control divided by the peak area of ​​the analytical internal standard) without running a standard curve. HRMS scans were performed over the appropriate m / z range to detect all possible metabolites.

[0165] Assay Conditions: This assay was performed using a single incubation (N=1). Test compounds were incubated at 37°C in a buffer containing 0.5 mg / mL liver microsomal protein. Reactions were initiated by the addition of cofactors and samples were taken at 0, 2, 4, 8, 16, 24, 36, and 48 hours. A positive control (5 μM testosterone) was incubated in parallel and samples were taken at 0, 2, 4, 8, 16, 24, 36, and 48 hours.

[0166] Assay quality control: A control compound, testosterone, was run in parallel to confirm (liver) microsomal enzyme activity. Fluorescence was used to confirm the addition of NADPH to the reaction mixture after the final time point. The T1 / 2 of the control met the acceptable internal standard.

[0167] Analytical methods:

[0168] Liquid chromatography column: Thermo BDS Hypersil C18 30 x 2.0 mm, 3 μm, with MP guard column; buffer: 25 mM formic acid buffer, pH 3.5; aqueous phase (A): 90% water, 10% buffer; organic phase (B): 90% acetonitrile, 10% buffer; flow rate: 300 μl / min; autosampler: injection volume: 10 μl. Gradient program: see Table 3.

[0169] Table 3 Gradient program

[0170] Time (minutes) ％A ％B 0.0 100 0 1.5 0 100 2.0 0 100 2.1 100 0 3.5 100 0

[0171] By using human liver microsomes, as described herein, Examples 4, 7, 9, and 10 exhibited metabolic half-lives greater than 24 hours, and Examples 3, 5, 8, and 11 exhibited metabolic half-lives between 16 and 24 hours, significantly greater than the 15-hour metabolic half-life of lazertinib. This relatively long metabolic half-life gives the present invention the potential to reduce medical dosages and extend dosing intervals.

[0172] The embodiments provided above are not intended to limit the scope of the present invention, nor are the steps described to limit their execution order. Any obvious improvements to the present invention made by those skilled in the art in combination with existing common knowledge shall fall within the scope of protection defined by the claims of the present invention.

Claims

1. A 2-arylaminopyrimidine compound having a structure represented by general formula (I) or a pharmaceutically acceptable salt thereof, in: X is CH; R 1 C 1-6 alkyl; R 2 is a 3- to 14-membered heterocyclic group; wherein the 3- to 14-membered heterocyclic group is optionally substituted by 1, 2 or 3 R's; R 3 C 1-6 alkyl; R 4 is H; R 5 and R 6 are independently selected from H, C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy; wherein the C 1-14 Alkyl, C 1-14 Alkoxy, C 3-6 Cycloalkyl and C 3-6 Cycloalkyloxy is optionally substituted with 1, 2 or 3 R""; R', R"" are independently selected from H, OH, C 1-6 alkyl; "Hetero" refers to a heteroatom or heteroatom group, and the "hetero" of the 3- to 14-membered heterocyclic group and the 3- to 6-membered heterocyclic group are independently selected from -N(R b )-、-NH-、-O-;R b Independently selected from C 1-6 alkyl.

2. The 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 Selected from C 1-3 alkyl.

3. The 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 2 It is a piperidine ring, piperazine ring or morpholine ring substituted with 1 to 3 R's.

4. The 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein R 5 and R 6 are independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl, wherein The C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl is optionally substituted with 1-3 R"".

5. The 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 5 and R 6 are independently selected from H, C 2-3 Alkyl; C 2-3 The alkyl group is optionally substituted with 1-3 R"".

6. The 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, characterized in that: The pharmaceutically acceptable salt is an inorganic salt or an organic salt. The inorganic salt includes hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, succinate, glutarate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, p-toluenesulfonate, and ascorbate.

7. A 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof, characterized in that: The 2-arylaminopyrimidine compound is specifically selected from:

8. A pharmaceutical composition comprising the 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, and a pharmaceutically acceptable carrier, excipient or diluent.

9. Use of the 2-arylaminopyrimidine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 in the preparation of a medicament for treating cancer mediated by EGFR activating or drug-resistant mutants in mammals.

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