Novel pyrazine compounds

Abstract

The application relates to a compound represented by a general formula (1) and a preparation method thereof, and the use of the compound represented by the general formula (1) and isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates thereof as EGFR inhibitors in the preparation of medicines for treating EGFR-related diseases such as tumors.

Description

[0001] This application claims priority to Chinese application CN202010486394.1, filed June 1, 2020; Chinese application CN202010947590.4, filed September 10, 2020; and Chinese patent application CN202110587528.3, filed May 27, 2021. The full text of the aforementioned Chinese applications is incorporated herein by reference. Technical Field

[0002] This invention relates to the field of medicinal chemistry, and more specifically, to a class of pyrazine compounds, their preparation methods, and the use of such compounds as EGFR inhibitors in the preparation of antitumor drugs. Background Technology

[0003] Lung cancer is one of the most common malignant tumors, with approximately 1.6 million new cases and 1.4 million deaths worldwide each year. Non-small cell lung cancer (NSCLC) accounts for about 80%-85% of all lung cancers (Nature, 2018, 553, 446-454).

[0004] The EGFR protein family is a class of protein kinases responsible for transmitting mitotic signals and playing a crucial role in growth and development. Extensive analysis and research of in vitro tumor cells, animal models, and human tumor samples have shown that mutations in EGFR family proteins lead to human tumor development and are a significant contributing factor to the occurrence and progression of various cancers. Therefore, targeting and inhibiting the activity of EGFR mutant proteins is an important approach to treating related tumors.

[0005] Studies show that EGFR gene mutations are found in approximately 12% to 47% of non-small cell lung cancers (NSCLC). In NSCLC, the two most common types of EGFR gene mutations are exon 19 deletion (del19) and L858 missense mutation in exon 21. These mutations lead to ligand-independent continuous activation of the EGFR protein. Although NSCLC patients with EGFR protein Del19 or L858R mutations are more sensitive to targeted therapy with EGFR protein kinase inhibitors (EGFR TKIs) such as erlotinib, gefitinib, afatinib, or osimertinib, achieving high clinical objective response rates (ORRs) of approximately 60-85%, this response typically does not last long, with most patients using first- or second-generation EGFR TKIs experiencing disease progression at around 11 months. Drug resistance analysis showed that in approximately 50-70% of resistant patients, the molecular mechanism of resistance was the acquisition of a second mutation in the EGFR gene, known as the T790M mutation (T790M+) (CancerDiscov. 2012, 2, 872-5). This secondary mutation caused first- and second-generation EGFR TKIs to lose their inhibitory activity against mutated tumor cells.

[0006] Osimertinib, a third-generation covalent EGFR TKI, was developed to treat tumors with EGFR del19 and L858R mutations, with or without the T790M mutation. While osimertinib has a high response rate against resistance caused by the T790M mutation, approximately 70% of patients eventually develop resistance, and the disease progresses again after about 10 months (Lung Cancer. 2017, 108, 228-231). Molecular mechanisms of resistance to third-generation EGFR TKIs have shown that a major resistance mechanism in approximately 20-40% of patients who relapse after osimertinib treatment is the acquisition of a third mutation in the EGFR gene, namely the C797S mutation. Furthermore, patients with the EGFR del19 / L858R T790MC797S mutation can no longer respond to first-, second-, or third-generation EGFR TKIs after treatment with a third-generation EGFR TKI. In 2015, Thrress et al. first reported an analysis of osimertinib resistance in 15 patients, finding that approximately 40% of the resistance was due to the C797S mutation (Nature Medicine, 2015, 21, 560-562). In 2017 at ASCO, Piotrowska and Zhou Caicun reported analyses of resistance in 23 and 99 patients respectively, both showing that approximately 22% of the resistance was caused by the C797S mutation. Therefore, targeting and inhibiting the EGFR del19 / L858R T790M C797S mutation could overcome osimertinib resistance. However, currently, there are no marketed EGFR TKIs that can inhibit the EGFR del19 / L858R T790M C797S mutant, making the research and development of fourth-generation EGFR TKIs to meet this unmet clinical need extremely urgent.

[0007] The EGFR del19 / L858R T790M C797S mutant, a newly emerging EGFR mutant after third-generation EGFR TKI treatment, is currently under-researched. Only a few fourth-generation EGFR TKIs have been reported to inhibit the EGFR del19 / L858R T790M C797S mutant. For example, Boehringer Ingelheim reported that a macrocyclic compound, BI-4020, possesses anti-EGFR del19 / L858R T790M C797S mutant activity and in vivo antitumor activity (J MedChem. 2019, 62, 10272-10293). Patent WO2019 / 015655 reports a class of arylphosphooxy compounds with anti-EGFR del19 / L858R T790M C797S mutant activity and in vivo antitumor activity. Its general formula A and representative compound B (Example 41 in the patent) have the following structures (please refer to the patent for the definition of each symbol in the formula):

[0008]

[0009] Currently, there is an urgent need to research and discover compounds with good activity against the EGFR del19 / L858R T790M C797S mutation. Summary of the Invention

[0010] The present invention aims to provide a compound of general formula (1) or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates:

[0011]

[0012] In general formula (1):

[0013] Y is a (3-11 quinone) heterocyclic alkyl, (C6-C14) aryl, or (5-10 quinone) heteroaryl, wherein the heterocyclic alkyl, aryl, or heteroaryl group may optionally be substituted with one or more of the following groups: -H, halogen, -R 4 -OR 4 -(CH2) n OR 4 -(CH2) n NR 4 R 5 -NR 4 R 5 -CN, -C(O)NR 4 R 5 -NR 5 C(O)R 4 -NR 5 S(O)2R 4-S(O) p R 4 -S(O)2NR 4 R 5 and -O-CH2-O-;

[0014] L 1 It can be -O- or -NH-;

[0015] X is (C6-C14) arylene or (5-11) heteroarylene, wherein the arylene and heteroarylene may optionally be substituted by one or more of the following groups: -H, halogen, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) alkoxy and (C1-C6) haloalkoxy;

[0016] R 1 For -H, halogen, -(CH2) n NR 6 R 7 -NR 6 R 7 -O(CH2) m NR 6 R 7 -N(R) 5 (CH2) m NR 6 R 7 (C1-C6)alkoxy, -CH2-(3-15)heterocyclic alkyl, or (3-15)heterocyclic alkyl, wherein the alkoxy and heterocyclic alkyl may optionally be substituted with one or more of the following groups: -H, -R 4 -(CH2) n NR 6 R 7 -NR 6 R 7 -O(CH2) m NR 6 R 7 -N(R) 5 (CH2) m NR 6 R 7 and -R 3 ;

[0017] L 2 It can be -O-, -NH-, or a chemical bond;

[0018] R 2 It is (C1-C6)alkyl, (C3-C14)cycloalkyl, (C6-C14)aryl, (3-4 member) heterocyclic alkyl, Or (6-11 quinary) heterocyclic alkyl; wherein the alkyl, cycloalkyl, aryl, heterocyclic alkyl, It may be optionally substituted with one or more of the following groups: -H, halogen, -R 4 -(CH2) n OR 4 -、-(CH2) n NR 4 R 5 -、-OR 4 -NR 4 R 5 -CN, -C(O)NR 4 R 5 -NR 5 C(O)R 4 -NR 5 S(O)2R 4 -S(O) p R 4 and -S(O)2NR 4 R 5 ;

[0019] R 3 It is a (3-11-membered) heterocyclic alkyl group, wherein the heterocyclic alkyl group may optionally be substituted by one or more of the following groups: -H, -CD3, -R 4 -OR 4 and -NR 4 R 5 ;

[0020] R 4 and R 5 Each is independently -H, (C1-C6)alkyl, or (C3-C14)cycloalkyl;

[0021] R 6 and R 7 Each is independently -H, (C1-C6)alkyl or (C3-C14)cycloalkyl, or R 6 and R 7 The N atom attached to it can together form a (3-11-membered) heterocyclic alkyl group, which may be optionally substituted by one or more of the following groups: -H, -CD3, halogen, -R 4 and -OR 4 ;

[0022] R 0 It is (C1-C6)alkyl or (C3-C14)cycloalkyl; and

[0023] p is an integer of 0, 1, or 2, n is an integer of 0, 1, 2, or 3, and m is an integer of 1, 2, or 3.

[0024] In another preferred embodiment, in the general formula (1), Y is a (5-6-membered) heterocyclic alkyl, phenyl, or (5-9-membered) heteroaryl, wherein the heterocyclic alkyl, phenyl, and heteroaryl may optionally be substituted with one or more of the following groups: -H, -F, -Cl, -Br, -CN, -OH, -OCH3, -NH2, -N(CH3)2, -NHCOCH3, -NHSO2CH3, -CH3, -CONH2, -CH2OH, and -O-CH2-O-.

[0025] In another preferred embodiment, Y in the general formula (1) is:

[0026]

[0027]

[0028] In another preferred embodiment, in the general formula (1), X is a phenylene or a 6-membered heteroaryl group, wherein the phenylene and heteroaryl group may optionally be substituted by one or more of the following groups: -H, -F, -CH3, -CH2CH3, -CH(CH3)2, -OCH3, -OCF2H and -OCF3.

[0029] In another preferred embodiment, X in the general formula (1) is:

[0030]

[0031] In another preferred embodiment, in the general formula (1), R 1 It is: -H, -N(CH3)2, -CH2-(6-11-membered)heterocyclic alkyl or (6-11-membered)heterocyclic alkyl, wherein the heterocyclic alkyl is: The heterocyclic alkyl group may optionally be substituted with one or more of the following groups: -H, -CH3, ... -N(CH3)2、 And -CD3.

[0032] In another preferred embodiment, in the general formula (1), R 1 for:

[0033] -H, -N(CH3)2,

[0034] In another preferred embodiment, wherein in the general formula (1), when L 2 When R is -NH-, 2 for:

[0035]

[0036] In another preferred embodiment, wherein in the general formula (1), when L 2 When R is -O-, 2 for:

[0037]

[0038] In another preferred embodiment, wherein in the general formula (1), when L 2 For: chemical bonds, R 2 for:

[0039]

[0040] In various embodiments, the representative compounds of the present invention have one of the following structures:

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059] Another object of the present invention is to provide a pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier, and a compound of general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates as active ingredients.

[0060] Another object of the present invention is to provide the use of the above-described compounds, or their isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, or the above-described pharmaceutical compositions, in the preparation of medicaments for treating EGFR mutation-related diseases.

[0061] Another object of the present invention is to provide a method for treating, modulating, and / or preventing diseases associated with EGFR mutant proteins, comprising administering to a subject a therapeutically effective amount of the above-described compound, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates, or solvates, or the above-described pharmaceutical compositions. Through the synthesis and careful study of various classes of novel compounds involving EGFR inhibition, the inventors discovered that in compounds of general formula (1), when Y is a heterocyclic alkyl, aromatic heterocyclic, or aryl group, the compound unexpectedly exhibits strong EGFR inhibition. del19 / T790M / C797S and EGFR L858R / T790M / C797S It inhibits activity and has high selectivity for wild-type EGFR WT.

[0062] It should be understood that the foregoing general description of the invention and the following detailed description are exemplary and illustrative, and are intended to provide further explanation of the claimed invention. Attached Figure Description

[0063] Figure 1 This is the tumor growth inhibition rate result of the in vivo drug efficacy study in mice in Example 4 of this invention;

[0064] Figure 2 This is the tumor growth inhibition rate result of the in vivo drug efficacy study in mice in Example 5 of this invention.

[0065] Compound Synthesis

[0066] The preparation method of the compound of general formula (1) of the present invention is described in detail below, but these specific methods do not constitute any limitation on the present invention.

[0067] The compounds of general formula (1) described above can be synthesized using standard synthetic techniques or known techniques combined with the methods described herein. Furthermore, the solvents, temperatures, and other reaction conditions mentioned herein can be varied. Starting materials used for the synthesis of the compounds can be obtained synthetically or from commercial sources. The compounds described herein and other related compounds with different substituents can be synthesized using known techniques and starting materials, including those discovered in March, ADVANCED ORGANIC CHEMISTRY 4. th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4 th Ed., Vols.A and B (Plenum 2000, 2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3 rd The method described in Ed. (Wiley 1999) can be used to prepare compounds by employing appropriate reagents and by introducing different groups into the molecular formulas provided herein.

[0068] On the one hand, the compounds described herein are prepared according to methods known in the art. However, the conditions of the method, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to the explanations below. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, such combinations being readily performed by those skilled in the art. On the other hand, the present invention also provides a method for preparing the compounds of the general formula (1) described herein, which is prepared using the following general reaction procedure 1 or general reaction procedure 2:

[0069] General reaction process 1

[0070]

[0071] Compounds of general formula (1) can be prepared according to general reaction procedure 1, wherein R 1 R 2 X, Y, L 1 and L 2 As defined above, H represents hydrogen, and B represents boric acid, borate ester, or trifluoroborate. As shown in general reaction flow 1, compound 1-1 reacts with formamide to produce compound 1-2, and compound 1-2 reacts with R... 1 -XL 1 -H reacts under alkaline conditions to form compound 1-3. Compound 1-3 undergoes a coupling reaction with YB to form compound 1-4. Compound 1-4 then reacts with R... 2 -L2 -H reacts under alkaline conditions to generate target compounds 1-5.

[0072] General reaction process 2

[0073]

[0074] Compounds of general formula (1) can be prepared according to general reaction procedure 2, wherein R 1 R 2 X, Y, L 1 and L 2 As defined above, H represents hydrogen. As shown in general reaction flow 2, compound 2-1 reacts with formamide to produce compound 2-2, and compound 2-2 reacts with R... 1 -XL 1 -H reacts under alkaline conditions to form compound 2-3, compound 2-3 and R 2 -L 2 -H reacts under alkaline conditions to form compound 2-4, and compound 2-4 reacts with YH under alkaline conditions to form the target compound 2-5.

[0075] Further forms of the compound

[0076] "Pharmaceutical acceptable" here means that a substance, such as a carrier or diluent, will not destroy the biological activity or properties of a compound and is relatively non-toxic. For example, when given to an individual, a substance will not cause unwanted biological effects or interact with any of its components in a harmful manner.

[0077] The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the administered organism and does not diminish the compound's biological activity and properties. In some specific respects, pharmaceutically acceptable salts are obtained by reacting compounds of formula (1) with acids, such as inorganic acids like hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, and phosphoric acid; organic acids like formic acid, acetic acid, propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid; and acidic amino acids like aspartic acid and glutamic acid.

[0078] It should be understood that references to pharmaceutically acceptable salts include solvent-added forms or crystalline forms, especially solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric solvents and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, etc. A hydrate is formed when the solvent is water, or an alcohol is formed when the solvent is ethanol. Solvates of compounds of general formula (1) are readily prepared or formed according to the methods described herein. For example, hydrates of compounds of general formula (1) are readily prepared by recrystallization from a mixture of water and organic solvents, including but not limited to tetrahydrofuran, acetone, ethanol, or methanol. Furthermore, the compounds mentioned herein can exist in both solvated and non-solvated forms. In summary, for the purposes of the compounds and methods provided herein, the solvated form is considered equivalent to the non-solvated form.

[0079] In other specific embodiments, compounds of general formula (1) are prepared in various forms, including but not limited to amorphous, pulverized, and nano-particle forms. Furthermore, compounds of general formula (1) include crystalline forms and can also be polymorphs. Polymorphs comprise different lattice arrangements of the same elemental composition of the compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability, and solubility. Different factors such as recrystallization solvents, crystallization rates, and storage temperatures may cause a single crystal form to dominate.

[0080] In another aspect, compounds of general formula (1) may possess a chiral center and / or axial chirality, and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers, and cis-trans isomers. Each chiral center or axial chirality will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of this invention. This invention means including all such isomeric forms of these compounds.

[0081] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 ( 125 I) and C-14 14 C). For example, deuterium can be used to replace hydrogen atoms to form deuterated compounds. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs generally have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug half-life in vivo. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0082] the term

[0083] Unless otherwise specified, the terms used in this application, including the specification and claims, are defined as follows. It must be noted that in the specification and appended claims, unless otherwise clearly indicated, the singular form "a" includes the plural meaning. Unless otherwise specified, conventional methods such as mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are used. In this application, unless otherwise specified, "or" or "and" refers to "and / or".

[0084] Unless otherwise specified, "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 14 carbon atoms. Lower alkyl groups containing 1 to 4 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, and tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyl groups, especially alkyl groups substituted with one or more halogens. Preferred alkyl groups are selected from CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, etc. i Pr、 n Pr、 i Bu、 n Bu or t Bu.

[0085] Unless otherwise specified, "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon double bond, including straight-chain or branched groups with 1 to 14 carbon atoms. Lower alkenyl groups containing 1 to 4 carbon atoms are preferred, such as vinyl, 1-propenyl, 1-butenyl, or 2-methylpropenyl.

[0086] Unless otherwise specified, "alkynyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon triple bond, including straight-chain and branched groups with 1 to 14 carbon atoms. Lower alkenyl groups containing 1 to 4 carbon atoms are preferred, such as ethynyl, 1-propynyl or 1-butynyl.

[0087] Unless otherwise specified, "cycloalkyl" refers to a 3- to 14-membered monocyclic aliphatic hydrocarbon group, in which one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexane, and cyclohexadiene.

[0088] Unless otherwise specified, "alkoxy" refers to an alkyl group bonded to the remainder of the molecule via an ether oxygen atom. Representative alkoxy groups are those having 1-6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy. As used herein, "alkoxy" includes unsubstituted and substituted alkoxy groups, particularly those substituted with one or more halogens. Preferred alkoxy groups are selected from OCH3, OCF3, CHF2O, CF3CH2O, etc.i- PrO, n- PrO, i- BuO、 n- BuO or t- BuO.

[0089] Unless otherwise specified, "aryl" refers to a hydrocarbon aromatic group, which can be monocyclic or polycyclic, such as a monocyclic aryl ring fused with one or more carbocyclic aromatic groups. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and phenanthrene.

[0090] Unless otherwise specified, "arylene" refers to a divalent aryl group as defined above. Examples of arylene groups include, but are not limited to, phenylene, naphthylene, and phenanthrene.

[0091] Unless otherwise specified, "heteroaryl" refers to an aromatic group containing one or more heteroatoms (O, S, or N), which may be monocyclic or polycyclic, such as a monocyclic heteroaryl ring fused with one or more carbocyclic aromatic groups or other monocyclic heterocyclic groups. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, furanyl, thiopheneyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrroleyl, indolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothiopheneyl, benzooxazolyl, benzopyridinyl, and pyrrolopyrimidinyl.

[0092] Unless otherwise specified, “hybrid aryl” refers to a divalent heteroaryl group as defined above.

[0093] Unless otherwise specified, "heterocyclic alkyl" refers to a saturated or partially unsaturated cyclic group containing one or more heteroatoms (O, S, or N), wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized as a ring atom. Unless otherwise stated, the cyclic system of "heterocyclic alkyl" can be monocyclic, bicyclic, spirocyclic, or polycyclic. "Heterocyclic alkyl" can be attached to the remainder of the molecule by more than one ring carbon or heteroatom. Examples of "heterocyclic alkyl" include, but are not limited to, pyrrolidine, piperidine, N-methylpiperidine, tetrahydroimidazolium, pyrazolidine, butyrolactam, valproic acid, imidazolinone, hydantoin, dioxolane, phthalimide, piperidine, pyrimidine-2,4(1H,3H)-dione, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrrolidine, thiaran, pyranone, tetrahydrofuran, tetrahydrothiophene, quinine ring, 2-azaspiro[3.3]heptane, etc.

[0094] Unless otherwise specified, "halogen" (or halogenated group) means fluorine, chlorine, bromine or iodine. The term "halogenated" (or "halogen substituted") appearing before the group name indicates that the group is partially or completely halogenated, that is, substituted by F, Cl, Br or I in any combination, preferably substituted by F or Cl.

[0095] "Optional" or "optionally" means that the event or condition described below may, but is not required, occur, and the description includes both the scenario in which the event or condition occurs and the scenario in which the event or condition does not occur.

[0096] The substituent "-O-CH2-O-" indicates that the two oxygen atoms in the substituent are connected to two adjacent carbon atoms of a heterocyclic alkyl, aryl, or heteroaryl group. For example:

[0097] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a single bond.

[0098] When one of the variables is selected as a chemical bond, it means that the two groups connected are directly linked. For example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0099] Specific pharmaceutical and medical terminology

[0100] The term “acceptable,” as used herein, means that a prescription component or active ingredient does not have an excessively harmful effect on health for general therapeutic purposes.

[0101] The terms “treatment,” “treatment process,” or “therapy” as used herein include alleviating, suppressing, or improving symptoms or conditions of a disease; suppressing the development of complications; improving or preventing underlying metabolic syndromes; suppressing the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; alleviating a disease or symptom; reducing complications arising from a disease or symptom; or preventing or treating signs arising from a disease or symptom. As used herein, a compound or pharmaceutical composition, when administered, may improve a disease, symptom, or condition, particularly by improving its severity, delaying its onset, slowing its progression, or reducing its duration. Whether administered regularly or intermittently, continuously or intermittently, it may be attributable to or related to the administration.

[0102] "Active ingredient" refers to the compound represented by general formula (1), and pharmaceutically acceptable inorganic or organic salts of compounds of general formula (1). The compounds of the present invention may contain one or more asymmetric centers (chiral centers or axial chirality), and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers. The asymmetric centers that may exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of the present invention. The present invention means including all such isomeric forms of these compounds.

[0103] The terms “compound,” “composition,” “agent,” or “medicine or medicament” may be used interchangeably here, and all refer to a compound or composition that, when applied to an individual (human or animal), can induce a desired pharmaceutical and / or physiological response through local and / or systemic action.

[0104] The term “administered, administering, or administration” here refers to the direct application of the compound or composition described herein, or the application of a prodrug, derivative, or analog of the active compound.

[0105] While the numerical ranges and parameters used to define the broader scope of this invention are approximate values, the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "approximately" may mean that the actual value falls within the acceptable standard error of the mean, as determined by those skilled in the art. Except for experimental examples, or unless explicitly stated otherwise, it is understood that all ranges, quantities, values, and percentages used herein (e.g., to describe material usage, duration, temperature, operating conditions, quantity ratios, and others similar) are modified with "approximately". Therefore, unless otherwise stated, the numerical parameters disclosed in this specification and the accompanying claims are approximate values ​​and are subject to change as needed. At a minimum, these numerical parameters should be understood as the indicated significant digits and values ​​obtained using general rounding.

[0106] Unless otherwise defined in this specification, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art. Furthermore, unless conflicting with the context, singular nouns used herein include their plural forms, and vice versa.

[0107] Therapeutic uses

[0108] This invention provides methods for treating diseases using the compounds or pharmaceutical compositions of this invention, including but not limited to conditions involving EGFR mutations (e.g., cancer).

[0109] In some embodiments, a method for treating cancer is provided, comprising administering to an individual in need an effective amount of any of the aforementioned protective formula (1) compounds. In some embodiments, the cancer is mediated by an EGFR mutation. In other embodiments, the cancer is lung cancer, pancreatic cancer, colon cancer, bladder cancer, brain cancer, breast cancer, urothelial carcinoma, prostate cancer, ovarian cancer, head and neck cancer, gastric cancer, mesothelioma, or metastatic cancer.

[0110] route of administration

[0111] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into various preparations, comprising, within a safe and effective range, the compounds of this invention or their pharmaceutically acceptable salts and pharmacologically acceptable excipients or carriers. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective range of the compound is determined based on the age, condition, and duration of treatment of the patient.

[0112] "Pharmaceutically acceptable excipients or carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), and emulsifiers (such as Tween). Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0113] When applying the compounds of this invention, they can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), or topically.

[0114] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0115] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0116] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0117] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0118] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0119] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0120] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0121] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0122] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 50–1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0123] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features. Detailed Implementation

[0124] The following description will elaborate on the specific aspects, characteristics, and advantages of the aforementioned compounds, methods, and pharmaceutical compositions, making the content of this invention readily apparent. It should be understood that the detailed descriptions and examples described below are specific embodiments and are for reference only. After reading this description, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by this application.

[0125] In all embodiments, 1 H-NMR was recorded using a Varian Mercury 400 NMR spectrometer, and chemical shifts are expressed as δ (ppm). Unless otherwise specified, the silica gel used for separation was 200-300 mesh, and all eluent ratios were by volume.

[0126] The following abbreviations are used in this invention: CDCl3 represents deuterated chloroform; CD3OD represents deuterated methanol; DMSO-d6 represents deuterated dimethyl sulfoxide; EtOAc represents ethyl acetate; Hexane represents n-hexane; MeCN represents acetonitrile; DCM represents dichloromethane; DIPEA represents diisopropylethylamine; NMP represents 1-methylpyrrolidone-2-one; Dioxane represents 1,4-dioxane; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; h represents hour. K3PO4 represents potassium phosphate; min represents minutes; MS represents mass spectrometry; NaH represents sodium hydride; NMR represents nuclear magnetic resonance; Pd2(dba)3 represents tris(dibenzylacetone)palladium; Pd(dppf)Cl2 represents 1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride; TFA(CF3COOH) represents trifluoroacetic acid; TLC represents thin-layer chromatography; THF represents tetrahydrofuran; Xantphos represents 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene.

[0127] Synthesis Method A:

[0128] The specific compounds 135 (5-((3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide) and their optical isomers (compounds 136, 137, 138 and 139) were synthesized using synthetic method A.

[0129]

[0130] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2):

[0131] 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-necked flask. The mixture was stirred and heated to 90 °C. Then, (NH4)2S2O8 (25 g, 109.1 mmol) solid was added in portions. The mixture was kept at 90 °C and stirred for 2 h. Then, K2S2O8 (30 g, 109.1 mmol) solid was added in portions again, and the mixture was kept at 90 °C and stirred for 20 h. LC-MS monitoring showed that the reaction contained product and the remaining reactants. Add EtOAc (150 mL) and water (300 mL) to the mixture, stir, separate the liquids, extract the aqueous phase with EtOAc (150 mL), combine the organic phases, wash and concentrate with saturated sodium chloride solution (150 mL), and purify the residue by column chromatography (EtOAc∶Hexane=0∶1 to 1∶5 to 1∶2) to obtain the product (1.82 g, yield: 10.5%), and recover the starting material (10.3 g, yield: 68.7%).

[0132] 1 H NMR (400MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0133] Step 2: Synthesis of compound 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_3):

[0134] 3,5-Dichloro-6-iodopyrazine-2-carboxamide (280 mg, 0.883 mmol), 4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (267 mg, 0.971 mmol), Dioxane (20 mL), and DIPEA (228 mg, 1.766 mmol) were added to a 50 mL single-necked flask. After purging the mixture with argon, the mixture was stirred and heated to reflux for 2 h. After the reaction was completed by LC-MS monitoring, the mixture was concentrated, and the residue was purified by column chromatography to obtain the product (368 mg, yield: 75%).

[0135] 1 H NMR (400MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J=3.8Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J=3.9Hz, 1H), 3.80-3.63 (m, 2H) ), 2.84-2.42 (m, 10H), 2.39 (ddt, J=11.4, 7.3, 3.7Hz, 1H), 1.96 (dt, J=12.2, 3.0Hz, 2H), 1.70 (qd, J=12.1, 4.0Hz, 2H); MS (ESI): 556[M+H] + .

[0136] Step 3: Synthesis of compound 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound int_4):

[0137] 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (167 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), phenylboronic acid (40.23 mg, 0.33 mmol), dioxane / H₂O (10 mL / 2 mL), and Pd(dppf)₂Cl₂ (22 mg) were added to a 50 mL single-necked flask. After purging the mixture with argon, the temperature was rapidly increased to 105 °C and the reaction was carried out for 30 min. The reaction was monitored by LC-MS until completion. After cooling, the mixture was purified by column chromatography to obtain the product (115 mg, yield: 75.6%).

[0138] 1 H NMR (400MHz, CDCl3) δ: 10.74 (s, 1H), 7.70 (d, J=6.5Hz, 3H), 7.56 (d, J=8.5Hz, 2H), 7.44 (p, J=6.8Hz, 3H), 6.94 (d, J=8.6Hz, 2H), 5.67 (s, 1H), 3.70 (d, J=11.9Hz, 2H), 2.83-2.55(m, 7H), 2.47(s, 3H), 2.40-2.32(m, 1H), 2.28(s, 3H), 1.93 (d, J=12.4Hz, 2H), 1.67 (tt, J=12.5, 6.8Hz, 2H); LC-MS: 506[M+H] + .

[0139] Step 4: Synthesis of compound 5-((3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 135):

[0140] Add 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazin-2-carboxamide (133.6 mg, 0.27 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (31 mg, 0.54 mmol), and DMSO (5 mL) to a 50 mL single-necked flask. Molecular sieve (200 mg powder) was added, and the mixture was stirred at room temperature for 15 min after purging with argon. Then, 3-aminocyclopentanol hydrochloride (45 mg, 0.32 mmol) was added, and the mixture was stirred and heated to 120 °C for 2 h after purging with argon. The reaction was monitored by LC-MS to ensure completion. After cooling, the mixture was purified by column chromatography to obtain the product (92 mg, yield: 59.7%).

[0141] MS(ESI): 571[M+H] + .

[0142] Using chiral separation technology, four optically pure chiral isomers can be obtained:

[0143]

[0144] 5-(((1R,3S)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 136):

[0145] 1 H NMR (400MHz, CDCl3) δ: 10.71 (s, 1H), 7.65-7.53 (m, 4H), 7.53-7.41 (m, 3H), 7.41-7.31 (m, 1H), 6.98-6.80 (m, 2H), 5 .21 (d, J=6.5Hz, 1H), 5.13 (s, 1H), 4.56 (h, J=7.1Hz, 1H), 4.39 (tt, J=5.8, 2.9Hz, 1H), 3.69 (d, J=12.0Hz, 2H), 2.80 -2.58 (m, 5H), 2.50 (s, 3H), 2.35 (d, J=9.8Hz, 1H), 2.30 (s, 3H), 2.29-2.22 (m, 1H), 2.18 (dd, J=13.8, 7.2Hz, 1H), 2. 06-1.96 (m, 1H), 1.92 (d, J=12.4Hz, 2H), 1.71-1.54 (m, 6H), 1.43 (ddd, J=13.0, 9.1, 6.6Hz, 1H); MS (ESI): 571[M+H] + .

[0146] 5-(((1R,3R)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 137):

[0147] 1H NMR (400MHz, CDCl3) δ: 10.78 (s, 1H), 7.60 (dd, J=8.6, 6.7Hz, 4H), 7.44 (t, J=7.5Hz, 3H ), 7.36 (t, J=7.3Hz, 1H), 6.91 (d, J=8.9Hz, 2H), 6.04 (d, J=7.4Hz, 1H), 5.10 (s, 1H), 4. 54 (d, J=7.8Hz, 1H), 4.44 (s, 1H), 3.68 (d, J=11.9Hz, 2H), 2.77-2.57 (m, 5H), 2.48 (s, 3 H), 2.36(s, 1H), 2.29(s, 3H), 2.17-1.99(m, 3H), 1.99-1.63(m, 9H); MS(ESI): 571[M+H] + .

[0148] 5-(((1S,3S)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 138):

[0149] 1 H NMR (400MHz, CDCl3) δ: 10.71 (s, 1H), 7.65-7.53 (m, 4H), 7.53-7.41 (m, 3H), 7.41-7.31 (m, 1H), 6.98-6.80 (m, 2H) , 5.21 (d, J=6.5Hz, 1H), 5.13 (s, 1H), 4.56 (h, J=7.1Hz, 1H), 4.39 (tt, J=5.8, 2.9Hz, 1H), 3.69 (d, J=12.0Hz, 2H), 2.80-2.58 (m, 5H), 2.50 (s, 3H), 2.35 (d, J=9.8Hz, 1H), 2.30 (s, 3H), 2.29-2.22 (m, 1H), 2.18 (dd, J=13.8, 7.2Hz, 1H), 2.06-1.96 (m, 1H), 1.92 (d, J=12.4Hz, 2H), 1.71-1.54 (m, 6H), 1.43 (ddd, J=13.0, 9.1, 6.6Hz, 1H); MS (ESI): 571[M+H] + .

[0150] 5-(((1S,3R)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 139):

[0151] 1H NMR (400MHz, CDCl3) δ: 10.78 (s, 1H), 7.60 (dd, J=8.6, 6.7Hz, 4H), 7.44 (t, J=7.5Hz, 3H ), 7.36 (t, J=7.3Hz, 1H), 6.91 (d, J=8.9Hz, 2H), 6.04 (d, J=7.4Hz, 1H), 5.10 (s, 1H), 4. 54 (d, J=7.8Hz, 1H), 4.44 (s, 1H), 3.68 (d, J=11.9Hz, 2H), 2.77-2.57 (m, 5H), 2.48 (s, 3 H), 2.36(s, 1H), 2.29(s, 3H), 2.17-1.99(m, 3H), 1.99-1.63(m, 9H); MS(ESI): 571[M+H] + .

[0152] The specific compound 39(3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(pyridin-4-yl)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide) was synthesized using synthetic method A.

[0153]

[0154] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2):

[0155] 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-necked flask. The mixture was stirred and heated to 90 °C. Then, (NH4)2S2O8 (25 g, 109.1 mmol) solid was added in portions. The mixture was kept at 90 °C and stirred for 2 h. Then, K2S2O8 (30 g, 109.1 mmol) solid was added in portions again, and the mixture was kept at 90 °C and stirred for 20 h. LC-MS monitoring showed that the reaction contained product and the remaining reactants. Add EtOAc (150 mL) and water (300 mL) to the mixture, stir, separate the liquids, extract the aqueous phase with EtOAc (150 mL), combine the organic phases, wash and concentrate with saturated sodium chloride solution (150 mL), and purify the residue by column chromatography (EtOAc∶Hexane=0∶1 to 1∶5 to 1∶2) to obtain the product (1.82 g, yield: 10.5%), and recover the starting material (10.3 g, yield: 68.7%).

[0156] 1 H NMR (400MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] +.

[0157] Step 2: Synthesis of compound 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_3):

[0158] 3,5-Dichloro-6-iodopyrazine-2-carboxamide (280 mg, 0.883 mmol), 4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (267 mg, 0.971 mmol), Dioxane (20 mL), and DIPEA (228 mg, 1.766 mmol) were added to a 50 mL single-necked flask. After purging the mixture with argon, the mixture was stirred and heated to reflux for 2 h. After the reaction was completed by LC-MS monitoring, the mixture was concentrated, and the residue was purified by column chromatography to obtain the product (368 mg, yield: 75%).

[0159] 1 H NMR (400MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J=3.8Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J=3.9Hz, 1H), 3.80-3.63 (m, 2H) ), 2.84-2.42 (m, 10H), 2.39 (ddt, J=11.4, 7.3, 3.7Hz, 1H), 1.96 (dt, J=12.2, 3.0Hz, 2H), 1.70 (qd, J=12.1, 4.0Hz, 2H); MS (ESI): 556[M+H] + .

[0160] Step 3: Synthesis of compound 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(pyridin-4-yl)pyrazin-2-carboxamide (compound int_6):

[0161] In a 50 mL single-necked flask, 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (166.76 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), (pyridin-4-yl)boric acid (40.56 mg, 0.33 mmol), Dioxane / H₂O (10 mL / 2 mL), and Pd(dppf)₂Cl₂ (22 mg) were added. After purging the mixture with argon, the temperature was rapidly increased to 105 °C and the reaction was carried out for 60 min. The reaction was monitored by LC-MS until completion. After cooling, the mixture was purified by column chromatography to obtain the product (126 mg, yield: 82.8%).

[0162] MS(ESI): 507[M+H] + .

[0163] Step 4: Synthesis of compound 3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(pyridin-4-yl)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide (compound 39)

[0164] Add 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(pyridin-4-yl)pyrazin-2-carboxamide (152.1 mg, 0.3 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (35 mg, 0.6 mmol), and DMSO (5 mL) to a 50 mL single-necked flask. Molecular sieve (200 mg powder) was added, and the mixture was stirred at room temperature for 15 min after purging with argon. Then, 3-tetrahydro-2H-pyran-4-amine (33.4 mg, 0.33 mmol) was added, and the mixture was stirred and heated to 120 °C for 2 h after purging with argon. The reaction was monitored by LC-MS to ensure completion. After cooling, the mixture was purified by column chromatography to obtain the product (130 mg, yield: 75.8%).

[0165] 1 H NMR (400MHz, CDCl3) δ10.86 (s, 1H), 8.71 (d, J=5.1Hz, 2H), 7.55 (t, J=6.8Hz, 4H) , 7.42 (s, 1H), 6.91 (d, J = 8.8Hz, 2H), 5.22 (s, 1H), 5.14 (d, J = 7.0Hz, 1H), 4.17 (m , 1H), 4.01 (d, J=11.7Hz, 2H), 3.69 (d, J=9.2Hz, 2H), 3.52 (t, J=11.6Hz, 2H), 2.6 9(m, 11H), 2.38(s, 3H), 2.09-1.96(m, 4H), 1.76-1.64(m, 4H); MS(ESI): 572[M+H] + .

[0166] The specific compound 55(3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(1H-pyrazol-3-yl)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide) was synthesized using synthetic method A.

[0167]

[0168] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2):

[0169] 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-necked flask. The mixture was stirred and heated to 90 °C. Then, (NH4)2S2O8 (25 g, 109.1 mmol) solid was added in portions. The mixture was kept at 90 °C and stirred for 2 h. Then, K2S2O8 (30 g, 109.1 mmol) solid was added in portions again, and the mixture was kept at 90 °C and stirred for 20 h. LC-MS monitoring showed that the reaction contained product and the remaining reactants. Add EtOAc (150 mL) and water (300 mL) to the mixture, stir, separate the liquids, extract the aqueous phase with EtOAc (150 mL), combine the organic phases, wash and concentrate with saturated sodium chloride solution (150 mL), and purify the residue by column chromatography (EtOAc∶Hexane=0∶1 to 1∶5 to 1∶2) to obtain the product (1.82 g, yield: 10.5%), and recover the starting material (10.3 g, yield: 68.7%).

[0170] 1 H NMR (400MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0171] Step 2: Synthesis of compound 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_3):

[0172] 3,5-Dichloro-6-iodopyrazine-2-carboxamide (280 mg, 0.883 mmol), 4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (267 mg, 0.971 mmol), Dioxane (20 mL), and DIPEA (228 mg, 1.766 mmol) were added to a 50 mL single-necked flask. After purging the mixture with argon, the mixture was stirred and heated to reflux for 2 h. After the reaction was completed by LC-MS monitoring, the mixture was concentrated, and the residue was purified by column chromatography to obtain the product (368 mg, yield: 75%).

[0173] 1H NMR (400MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J=3.8Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J=3.9Hz, 1H), 3.80-3.63 (m, 2H) ), 2.84-2.42 (m, 10H), 2.39 (ddt, J=11.4, 7.3, 3.7Hz, 1H), 1.96 (dt, J=12.2, 3.0Hz, 2H), 1.70 (qd, J=12.1, 4.0Hz, 2H); MS (ESI): 556[M+H] + .

[0174] Step 3: Synthesis of compound 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(1H-pyrazol-3-yl)pyrazin-2-carboxamide (compound int_7):

[0175] In a 50 mL single-necked flask, 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (166.76 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), (1H-pyrazol-3-yl)boric acid (37 mg, 0.33 mmol), Dioxane / H₂O (10 mL / 2 mL), and Pd(dppf)₂Cl₂ (22 mg) were added. After purging the mixture with argon, the temperature was rapidly increased to 105 °C and the reaction was carried out for 60 min. The reaction was monitored by LC-MS until completion. After cooling, the mixture was purified by column chromatography to obtain the product (119 mg, yield: 80%).

[0176] MS(ESI): 496 [M+H] + .

[0177] Step 4: Synthesis of compound 3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(1H-pyrazol-3-yl)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide (compound 55)

[0178] Add 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(1H-pyrazol-3-yl)pyrazin-2-carboxamide (148.8 mg, 0.3 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (35 mg, 0.6 mmol), and DMSO (5 mL) to a 50 mL single-necked flask. Molecular sieve (200 mg powder) was added, and the mixture was stirred at room temperature for 15 min after purging with argon. Then, 3-tetrahydro-2H-pyran-4-amine (33.4 mg, 0.33 mmol) was added, and the mixture was stirred and heated to 120 °C for 2 h after purging with argon. The reaction was monitored by LC-MS to ensure completion. After cooling, the mixture was purified by column chromatography to obtain the product (115 mg, yield: 68.3%).

[0179] Compound 55 fumarate: 1 H NMR (400MHz, DMSO-d6) δ13.05 (s, 1H), 11.21 (s, 1H), 8.90 (s, 1H), 8.00-7.92 (m, 1H), 7.81 (d, J=2.4Hz, 1H), 7.55-7.47 (m, 2H), 7.31 (d, J=2.8Hz, 1H), 7. 21 (d, J=2.4Hz, 1H), 6.89 (d, J=8.9Hz, 2H), 6.54 (s, 4H), 4.11 (m, J=6.2Hz, 1H), 3.89 (dt, J=1 1.5, 3.7Hz, 2H), 3.62 (d, J=11.8Hz, 2H), 3.49 (td, J=11.5, 2.4Hz, 2H), 2.88-2.52 (m, 9H), 2. 42(s, 3H), 2.10-1.99(m, 2H), 1.84(d, J=11.1Hz, 2H), 1.61-1.40(m, 4H); MS(ESI): 561[M+H] + .

[0180] The specific compound 511 (6-(1H-indol-4-yl)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide) was synthesized using synthetic method A.

[0181]

[0182] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2):

[0183] 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-necked flask. The mixture was stirred and heated to 90 °C. Then, (NH4)2S2O8 (25 g, 109.1 mmol) solid was added in portions. The mixture was kept at 90 °C and stirred for 2 h. Then, K2S2O8 (30 g, 109.1 mmol) solid was added in portions again, and the mixture was kept at 90 °C and stirred for 20 h. LC-MS monitoring showed that the reaction contained product and the remaining reactants. Add EtOAc (150 mL) and water (300 mL) to the mixture, stir, separate the liquids, extract the aqueous phase with EtOAc (150 mL), combine the organic phases, wash and concentrate with saturated sodium chloride solution (150 mL), and purify the residue by column chromatography (EtOAc∶Hexane=0∶1 to 1∶5 to 1∶2) to obtain the product (1.82 g, yield: 10.5%), and recover the starting material (10.3 g, yield: 68.7%).

[0184] 1 H NMR (400MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0185] Step 2: Synthesis of compound 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_3):

[0186] 3,5-Dichloro-6-iodopyrazine-2-carboxamide (280 mg, 0.883 mmol), 4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (267 mg, 0.971 mmol), Dioxane (20 mL), and DIPEA (228 mg, 1.766 mmol) were added to a 50 mL single-necked flask. After purging the mixture with argon, the mixture was stirred and heated to reflux for 2 h. After the reaction was completed by LC-MS monitoring, the mixture was concentrated, and the residue was purified by column chromatography to obtain the product (368 mg, yield: 75%).

[0187] 1H NMR (400MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J=3.8Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J=3.9Hz, 1H), 3.80-3.63 (m, 2H) ), 2.84-2.42 (m, 10H), 2.39 (ddt, J=11.4, 7.3, 3.7Hz, 1H), 1.96 (dt, J=12.2, 3.0Hz, 2H), 1.70 (qd, J=12.1, 4.0Hz, 2H); MS (ESI): 556[M+H] + .

[0188] Step 3: Synthesis of compound 5-chloro-6-(1H-indol-4-yl)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_8):

[0189] 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (166.76 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), (1H-indol-4-yl)boric acid (53.12 mg, 0.33 mmol), Dioxane / H₂O (10 mL / 2 mL), and Pd(dppf)₂Cl₂ (22 mg) were added to a 50 mL single-necked flask. After purging the mixture with argon, the temperature was rapidly increased to 105 °C and the reaction was carried out for 60 min. The reaction was monitored by LC-MS until completion. After cooling, the mixture was purified by column chromatography to obtain the product (131 mg, yield: 80%).

[0190] MS(ESI): 545 [M+H] + .

[0191] Step 4: Synthesis of compound 6-(1H-indol-4-yl)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide (compound 511)

[0192] Add 5-chloro-6-(1H-indol-4-yl)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (164 mg, 0.3 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (35 mg, 0.6 mmol), and DMSO (5 mL) to a 50 mL single-necked flask. Molecular sieve (200 mg powder) was added, and the mixture was stirred at room temperature for 15 min after purging with argon. Then, 3-tetrahydro-2H-pyran-4-amine (33.4 mg, 0.33 mmol) was added, and the mixture was stirred and heated to 120 °C for 2 h after purging with argon. The reaction was monitored by LC-MS to ensure completion. After cooling, the mixture was purified by column chromatography to obtain the product (127 mg, yield: 69.4%).

[0193] 1 H NMR (400MHz, CDCl3) δ10.81 (s, 1H), 8.39 (s, 1H), 7.62 (d, J=8.9Hz, 2H), 7.55-7.45 (m , 2H), 7.30 (td, J=6.3, 5.7, 4.0Hz, 3H), 6.92 (d, J=8.9Hz, 2H), 6.54 (d, J=2.8Hz, 1H), 5.25 (d, J=7.2Hz, 1H), 5.13 (s, 1H), 4.23-4.14 (m, 1H), 3.96 (d, J=11.7Hz, 2H), 3.69 (d, J=11.9Hz, 2H), 3.56-3.47 (m, 2H), 2.77 -2.37 (m, 11H), 2.32 (s, 3H), 2.00 (dd, J=26.0, 11.4Hz, 4H), 1.72 (dd, J=11.8, 3.8Hz, 2H), 1.49-1.42 (m, 2H); MS (ESI): 610[M+H] + .

[0194] Synthesis Method B:

[0195] The specific compound 19 (5-methoxy-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide) was synthesized using synthetic method B:

[0196]

[0197] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2):

[0198] 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-necked flask. The mixture was stirred and heated to 90 °C. Then, (NH4)2S2O8 (25 g, 109.1 mmol) solid was added in portions. The mixture was kept at 90 °C and stirred for 2 h. Then, K2S2O8 (30 g, 109.1 mmol) solid was added in portions again, and the mixture was kept at 90 °C and stirred for 20 h. LC-MS monitoring showed that the reaction contained product and the remaining reactants. Add EtOAc (150 mL) and water (300 mL) to the mixture, stir, separate the liquids, extract the aqueous phase with EtOAc (150 mL), combine the organic phases, wash and concentrate with saturated sodium chloride solution (150 mL), and purify the residue by column chromatography (EtOAc∶Hexane=0∶1 to 1∶5 to 1∶2) to obtain the product (1.82 g, yield: 10.5%), and recover the starting material (10.3 g, yield: 68.7%).

[0199] 1 H NMR (400MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0200] Step 2: Synthesis of compound 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_3):

[0201] 3,5-Dichloro-6-iodopyrazine-2-carboxamide (280 mg, 0.883 mmol), 4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (267 mg, 0.971 mmol), Dioxane (20 mL), and DIPEA (228 mg, 1.766 mmol) were added to a 50 mL single-necked flask. After purging the mixture with argon, the mixture was stirred and heated to reflux for 2 h. After the reaction was completed by LC-MS monitoring, the mixture was concentrated, and the residue was purified by column chromatography to obtain the product (368 mg, yield: 75%).

[0202] 1H NMR (400MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J=3.8Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J=3.9Hz, 1H), 3.80-3.63 (m, 2H) ), 2.84-2.42 (m, 10H), 2.39 (ddt, J=11.4, 7.3, 3.7Hz, 1H), 1.96 (dt, J=12.2, 3.0Hz, 2H), 1.70 (qd, J=12.1, 4.0Hz, 2H); MS (ESI): 556[M+H] + .

[0203] Step 3: Synthesis of compound 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound int_4):

[0204] 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (167 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), phenylboronic acid (40.23 mg, 0.33 mmol), dioxane / H₂O (10 mL / 2 mL), and Pd(dppf)₂Cl₂ (22 mg) were added to a 50 mL single-necked flask. After purging the mixture with argon, the temperature was rapidly increased to 105 °C and the reaction was carried out for 30 min. The reaction was monitored by LC-MS until completion. After cooling, the mixture was purified by column chromatography to obtain the product (115 mg, yield: 75.6%).

[0205] 1 H NMR (400MHz, CDCl3) δ: 10.74 (s, 1H), 7.70 (d, J=6.5Hz, 3H), 7.56 (d, J=8.5Hz, 2H), 7.44 (p, J=6.8Hz, 3H), 6.94 (d, J=8.6Hz, 2H), 5.67 (s, 1H), 3.70 (d, J=11.9Hz, 2H), 2.83-2.55(m, 7H), 2.47(s, 3H), 2.40-2.32(m, 1H), 2.28(s, 3H), 1.93 (d, J=12.4Hz, 2H), 1.67 (tt, J=12.5, 6.8Hz, 2H); LC-MS: 506[M+H] + .

[0206] Step 4: Synthesis of compound 5-methoxy-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 19):

[0207] Add 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (50 mg, 0.10 mmol) and DMF (5 mL) to a 50 mL single-necked flask. Molecular sieve (200 mg, powder) was added, and after purging the mixture with argon, the mixture was stirred at room temperature for 15 min. Then, sodium methoxide (16 mg, 0.3 mmol) was added, and after purging the mixture with argon, the mixture was stirred and heated to 80 °C for 2 h. The reaction was monitored by LC-MS until it was complete. After cooling the mixture, the product was purified by column chromatography (21 mg, yield: 42%).

[0208] 1 H NMR (400MHz, CDCl3) δ: 10.81 (s, 1H), 7.99-7.87 (m, 2H), 7.65 (d, J=4.4Hz, 1H), 7.61-7 .55 (m, 2H), 7.41 (d, J=7.8Hz, 2H), 6.94 (dd, J=9.0, 3.6Hz, 2H), 5.42-5.33 (m, 1H), 4.05 (s, 3H), 3.70 (d, J=12.0Hz, 2H), 2.78-2.41 (m, 11H), 2.41-2.32 (m, 1H), 2.30 (s , 3H), 1.94 (d, J=12.4Hz, 2H), 1.69 (qd, J=11.8, 3.7Hz, 2H); MS (ESI): 502[M+H] + .

[0209] Synthesis method C:

[0210] The specific compound 116(3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenyl-5-((tetrahydro-2H-pyran-4-yl)oxo)pyrazin-2-carboxamide) was synthesized using synthetic method C.

[0211]

[0212] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2):

[0213] 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-necked flask. The mixture was stirred and heated to 90 °C. Then, (NH4)2S2O8 (25 g, 109.1 mmol) solid was added in portions. The mixture was kept at 90 °C and stirred for 2 h. Then, K2S2O8 (30 g, 109.1 mmol) solid was added in portions again, and the mixture was kept at 90 °C and stirred for 20 h. LC-MS monitoring showed that the reaction contained product and the remaining reactants. Add EtOAc (150 mL) and water (300 mL) to the mixture, stir, separate the liquids, extract the aqueous phase with EtOAc (150 mL), combine the organic phases, wash and concentrate with saturated sodium chloride solution (150 mL), and purify the residue by column chromatography (EtOAc∶Hexane=0∶1 to 1∶5 to 1∶2) to obtain the product (1.82 g, yield: 10.5%), and recover the starting material (10.3 g, yield: 68.7%).

[0214] 1 H NMR (400MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0215] Step 2: Synthesis of compound 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (compound int_3):

[0216] 3,5-Dichloro-6-iodopyrazine-2-carboxamide (280 mg, 0.883 mmol), 4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (267 mg, 0.971 mmol), Dioxane (20 mL), and DIPEA (228 mg, 1.766 mmol) were added to a 50 mL single-necked flask. After purging the mixture with argon, the mixture was stirred and heated to reflux for 2 h. After the reaction was completed by LC-MS monitoring, the mixture was concentrated, and the residue was purified by column chromatography to obtain the product (368 mg, yield: 75%).

[0217] 1H NMR (400MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J=3.8Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J=3.9Hz, 1H), 3.80-3.63 (m, 2H) ), 2.84-2.42 (m, 10H), 2.39 (ddt, J=11.4, 7.3, 3.7Hz, 1H), 1.96 (dt, J=12.2, 3.0Hz, 2H), 1.70 (qd, J=12.1, 4.0Hz, 2H); MS (ESI): 556[M+H] + .

[0218] Step 3: Synthesis of compound 6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide (compound int_5):

[0219] Add 5-chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazin-2-carboxamide (150 mg, 0.27 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (31 mg, 0.54 mmol), and DMSO (5 mL) to a 50 mL single-necked flask. Molecular sieve (200 mg, powder) was mixed with argon gas and stirred at room temperature for 15 min. Then, 3-tetrahydro-2H-pyran-4-amine (32 mg, 0.32 mmol) was added, and the mixture was purged with argon gas and stirred until heated to 120 °C for 2 h. The reaction was monitored by LC-MS until completion. After cooling, the mixture was purified by column chromatography to obtain the product (110 mg, yield: 65.7%). 1 H NMR (400MHz, CDCl3) δ: 10.64 (s, 1H), 7.62-7.39 (m, 2H), 7.21 (s, 1H), 6.96-6 .76 (m, 2H), 5.41-5.12 (m, 2H), 4.03 (dq, J=11.4, 3.7Hz, 3H), 3.67 (d, J=12.0 Hz, 2H), 3.51 (td, J=11.6, 2.2Hz, 2H), 2.84-2.50 (m, 10H), 2.44 (d, J=11.4Hz , 1H), 2.37(s, 3H), 2.11-1.89(m, 4H), 1.78-1.51(m, 4H); MS(ESI): 621[M+H] + .

[0220] Step 4: Synthesis of compound 3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(piperidin-1-yl)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide (compound 116):

[0221] 6-Iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-carboxamide (57 mg, 0.10 mmol), piperidine (34 mg, 0.40 mmol), anhydrous cesium fluoride (45 mg, 0.30 mmol), and NMP (5 ml) were added to a 10 ml microwave reactor. The mixture was heated to 180 °C and stirred for 8 h. After the reaction was completed, the mixture was cooled by LC-MS and purified by column chromatography to obtain the product (32 mg, yield: 57.4%).

[0222] 1 H NMR (400MHz, CDCl3) δ: 10.57 (s, 1H), 7.55-7.48 (m, 2H), 6.90-6.82 (m, 2H), 5.51 (d, J=7. 3Hz, 1H), 5.06 (s, 1H), 4.13-4.04 (m, 1H), 4.02-3.96 (m, 2H), 3.64 (d, J=11.7Hz, 2H), 3.5 5(td, J=11.5, 2.3Hz, 2H), 2.87(t, J=5.3Hz, 4H), 2.71-2.58(m, 5H), 2.48(s, 3H), 2.36(t , J=11.4Hz, 1H), 2.29 (s, 3H), 2.06 (d, J=13.1Hz, 2H), 1.93 (d, J=12.3Hz, 2H), 1.73-1.52 (m, 12H); MS(ESI): 578[M+H] + .

[0223] Example 1: Synthesis of Compound 1-645

[0224] Using synthesis method A, synthesis method B, or synthesis method C, and with different raw materials, the target compounds 1-18, 20-115, 117-135, and 140-645 in Table 2 can be obtained.

[0225] The LC-MS analysis method is as follows:

[0226] Instrument: Agilent 6125B

[0227] Chromatographic column: Core-shell 2.7um 4.3*50mm

[0228] Column temperature: 30℃

[0229] Wavelength: 254nm / 214nm

[0230] Mobile phase A: H2O (0.1% formic acid)

[0231] Mobile phase B: Acetonitrile (0.1% formic acid)

[0232] gradient:

[0233] Table 1

[0234] Time (min) Flow rate (mL / min) Mobile phase B% Mobile phase A% 0 2 5 95 0.1 2 5 95 2.2 2 95 5 2.7 2 95 5 2.71 2 5 95 3.2 2 5 95

[0235] Table 2

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318] Table 3. NMR data of some compounds in Table 2

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330]

[0331]

[0332] Example 2: Detection of the inhibitory activity of the compounds of the present invention against EGFR (del19 / T790M / C797S), EGFR (L858R / T790M / C797S), or EGFR (WT) enzymes.

[0333] The inhibitory effects of the compounds on the activities of EGFR (del19 / T790M / C797S), EGFR (L858R / T790M / C797S), or EGFR (WT) enzymes were determined using the HTRF method. Details are as follows.

[0334] After incubating WT or mutant EGFR protein with serially diluted compounds at 28°C for 10 minutes, biotin-labeled universal tyrosine kinase substrate (TK) and ATP were added, and the reaction was allowed to proceed at room temperature for 40 minutes. Upon termination of the reaction, Eu3+-Cryptate-labeled antibody against TK and streptavidin-XL665 were added, and the mixture was incubated at room temperature for 60 minutes. The phosphorylation level of the TK substrate was quantified by calculating the 665 / 615 ratio based on the emission at 615 nm and 665 nm. The percentage of inhibition and IC50 of the compound were calculated compared to the control group. 50 The results are shown in Table 4 below.

[0335] Table 4. Inhibitory activity of the compounds of the present invention against EGFR (del19 / T790M / C797S), EGFR (L858R / T790M / C797S), or EGFR (WT).

[0336]

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343] + indicates an inhibition rate of 20% or less.

[0344] ++ indicates an inhibition rate of 20% to 50%.

[0345] +++ indicates an inhibition rate greater than 50%.

[0346] ND indicates that the activity was not measured.

[0347] As shown in Table 4, the compounds of the present invention have good inhibitory activity against the enzyme activities of EGFR(del19 / T790M / C797S) and EGFR(L858R / T790M / C797S), and good selectivity for EGFR(WT).

[0348] Example 3: The effect of the compound of the present invention on Ba / F3 (EGFR) del19 / T790M / C797S Antiproliferative activity of triple mutant cells and A431 (EGFR WT) cells

[0349] 3000 Ba / F3 cells carrying EGFR (del19 / T790M / C797S) or 2000 A431 cells were seeded in 384-well plates. After one day of growth, serially diluted compounds were added (maximum 500 nM for Ba / F3 cells, maximum 10 μM for A431 cells). Three days after compound addition, Cell Titer Glow was added to evaluate cell growth, and the percentage of cell growth inhibition and IC50 of the compounds were calculated. 50 The values ​​are shown in Table 5 below.

[0350] Table 5. Effects of the compounds of the present invention on Ba / F3 (EGFR) del19 / T790M / C797S Antiproliferative activity of triple mutant cells and A431 wild-type (EGFRWT) cells

[0351]

[0352]

[0353]

[0354]

[0355] As shown in Table 5, the vast majority of compounds in this invention are effective against Ba / F3 (EGFR). del19 / T790M / C797S The antiproliferative activity of the triple mutant cells was less than 100 nM, while Gilteritinib showed antiproliferative activity against Ba / F3 (EGFR) cells. del19 / T790M / C797S The antiproliferative activity of the triple mutant cells is greater than 500 nM, indicating that when Y is aryl, heteroaryl, or heterocyclic alkyl, the compound exhibits strong Ba / F3 (EGFR) activity. del19 / T790M / C797S Antiproliferative activity of triple mutant cells.

[0356] Example 4: In vivo efficacy study - mouse H1975 subcutaneous xenograft model

[0357] BALB / c nude mice were subcutaneously injected with 5*10 slits on the left back. 6H1975 cells carrying the EGFR T790M mutation were cultured until the tumor grew to 100-150 mm. 3 After randomization, participants were administered compound 511 via gavage: Group 1: solvent control group; Group 2: compound 511 (60 mg / kg); Group 3: compound 511 (80 mg / kg), once daily. Tumor volume was measured twice weekly and at the dosing endpoint. The tumor growth inhibition rate (TGI) was calculated as: TGI = 1 - (tumor volume on day 28 in the treated group - tumor volume on day 1 in the treated group) / (tumor volume on day 28 in the solvent control group - tumor volume on day 1 in the treated group). Results are shown below. Figure 1 See Table 6.

[0358] Table 6. Growth inhibition rate of H1975 subcutaneous xenograft tumors in mice

[0359] compound dose TGI Comparison not applicable not applicable Compound 511 60mg / kg 99％ Compound 511 80mg / kg 104％

[0360] Depend on Figure 1 As shown in Table 6, compound 511 inhibited tumor growth in a subcutaneous xenograft model of H1975 mice carrying the EGFR T790M mutation at doses of 60 mg / kg and 80 mg / kg.

[0361] Example 5: In vivo efficacy study - mouse PC9 (EGFR Del19 / T790M / C797S) subcutaneous xenograft model

[0362] BALB / c nude mice were subcutaneously injected with 5 x 10 dextrose tablets on the left back. 6 PC9 cells carrying EGFR Del19 / T790M / C797S were used until the tumor grew to 100-150 mm. 3 After randomization, participants were administered compound 511 via gavage: Group 1: solvent control group; Group 2: compound 511 (60 mg / kg); Group 3: compound 511 (80 mg / kg), once daily. Tumor volume was measured twice weekly and at the dosing endpoint. The tumor growth inhibition rate (TGI) was calculated as: TGI = 1 - (tumor volume on day 28 in the treated group - tumor volume on day 1 in the treated group) / (tumor volume on day 28 in the solvent control group - tumor volume on day 1 in the treated group). Results are shown below. Figure 2 .

[0363] Table 7. Growth inhibition rate of subcutaneous xenografts of mouse PC9 (EGFR Del19 / T790M / C797S)

[0364] compound dose TGI Comparison not applicable not applicable Compound 511 60mg / kg 87.41％ Compound 511 80mg / kg 93.17％

[0365] Depend on Figure 2As shown in Table 7, compound 511 inhibited tumor growth in a PC9 mouse subcutaneous xenograft model carrying the EGFRDel19 / T790M / C797S mutation at doses of 60 mg / kg and 80 mg / kg.

Claims

1. A compound with a structure as shown in general formula (1), or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof: In general formula (1): Y is (C6-C14) aryl or (5-10-membered) heteroaryl; the aryl group is phenyl or naphthyl, wherein the aryl and heteroaryl groups may optionally be substituted by one or more of the following groups: halogen, -R 4 -OR 4 -NR 4 R 5 In -CN;Y, R 4 H or (C1-C6) alkyl, R 5 It is H or (C1-C6) alkyl; L 1 It is -NH-; X is (C6-C14) arylene; the arylene is phenylene, wherein the arylene may optionally be substituted by one or more of the following groups: -H, halogen, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) alkoxy and (C1-C6) haloalkoxy; R 1 It is: -H, -N(CH3)2, -CH2-(6-11-membered)heterocyclic alkyl or (6-11-membered)heterocyclic alkyl, wherein the heterocyclic alkyl is: , , , , , , , , , , , , The heterocyclic alkyl group may optionally be substituted with one or more of the following groups: -H, -CH3, -N(CH3)2, , , , , , , , , and -CD3; L 2 When it is -NH-; R 2 for , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ; L 2 When it is -O-; R 2 (C1-C6)alkyl, (C3-C14)cycloalkyl, (3-4 member) heterocyclic alkyl, , Or a 6-membered heterocyclic alkyl group; wherein the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, and wherein the (C1-C6)alkyl, (C3-C14)cycloalkyl, (3-4-membered) heterocyclic alkyl, and 6-membered heterocyclic alkyl group are optionally substituted with one or more of the following groups: halogen, -R 4 -OR 4 and -S(O) p R 4 ;R 2 In the middle, R 4 It is -H or (C1-C6) alkyl; p is 2, and m is an integer of 1, 2, or 3.

2. A compound with a structure as shown in general formula (1), or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof: In general formula (1): Y is a (5-6-membered) heterocyclic alkyl, phenyl, or (5-9-membered) heteroaryl, wherein the (5-6-membered) heterocyclic alkyl, phenyl, or heteroaryl may optionally be substituted by one or more of the following groups: -H, -F, -Cl, -Br, -CN, -OH, -OCH3, -NH2, -N(CH3)2, -NHCOCH3, -NHSO2CH3, -CH3, -CONH2, or -CH2OH; L 1 It is -NH-; X is (C6-C14) arylene; the arylene is phenylene, wherein the arylene may optionally be substituted by one or more of the following groups: -H, halogen, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) alkoxy and (C1-C6) haloalkoxy; R 1 It is -H, -N(CH3)2, -CH2-(6-11-membered) heterocyclic alkyl or (6-11-membered) heterocyclic alkyl, wherein the heterocyclic alkyl is: , , , , , , , , , , , , The heterocyclic alkyl group may optionally be substituted with one or more of the following groups: -H, -CH3, -N(CH3)2, , , , , , , , , and -CD3; L 2 When it is -NH-; R 2 for , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or ; L 2 When it is -O-; R 2 It is (C1-C6)alkyl, (C3-C14)cycloalkyl, (C6-C14)aryl, (3-4 member) heterocyclic alkyl, , , , Or a 6-membered heterocyclic alkyl group; wherein the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, and the aryl group is phenyl or naphthyl, wherein the (C1-C6)alkyl, (C3-C14)cycloalkyl, (C6-C14) aryl, (3-4-membered) heterocyclic alkyl, and 6-membered heterocyclic alkyl group may optionally be substituted by one or more of the following groups: halogen, -R 4 -OR 4 or -S(O) p R 4 ;R 2 In the middle, R 4 It is -H or (C1-C6) alkyl; L 2 When it is a chemical bond; R 2 for , , , , , or ; R 0 It is (C1-C6) alkyl; and p is 2, and m is an integer of 1, 2, or 3; The heterocyclic alkyl group is a saturated or partially unsaturated cyclic system group containing one or more heteroatoms, wherein the heteroatoms are O, S or N.

3. The compound of claim 1 or a single enantiomer, a single diastereomer or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), Y is a phenyl or (5-9) heteroaryl group, wherein the phenyl and heteroaryl group may optionally be substituted with one or more of the following groups: -H, -F, -Cl, -Br, -CN, -OH, -OCH3, -NH2, -N(CH3)2 and -CH3.

4. The compound of claim 3 or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), Y is: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

5. The compound of claim 2 or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), Y is: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

6. The compound of any one of claims 1-5, or a single enantiomer, a non-single enantiomer, or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), X is a phenylene, wherein the phenylene may optionally be substituted with one or more of the following groups: -H, -F, -CH3, -CH2CH3, -CH(CH3)2, -OCH3, -OCF2H and -OCF3.

7. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), X is: , , , , , , , , , , , , , , , or .

8. The compound of claim 1 or 2, or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), R 1 for: -H, -N(CH3)2, , , , , , , , , , , , , , or .

9. The compound of claim 1 or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), when L 2 When it is -O-, R 2 for: , , , or .

10. The compound of claim 2 or a pharmaceutically acceptable salt thereof, wherein in the general formula (1), when L 2 When it is -O-, R 2 for: , , , , , , , , , or .

11. A compound as described below, or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof, wherein said compound has one of the following structures: 。 12. A pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier, and a compound as described in any one of claims 1-11, or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof as an active ingredient.

13. The use of a compound as described in any one of claims 1-11, or a single enantiomer, a single diastereomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 12, in the preparation of a medicament for treating lung cancer.

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