Novel pyrazine compounds

Abstract

The present invention relates to compounds of general formula (1) and processes for their preparation, as well as the use of compounds of general formula (1) and their isomers, crystalline forms, pharmaceutically acceptable salts, hydrates or solvates as EGFR inhibitors in the preparation of medicaments for EGFR-related diseases such as tumors. [Formula 1] TIFF2023528859000191.tif30151

Description

[Technical Field]

[0001] This application claims priority to Chinese Patent Application No. 202010486394.1, filed on June 1, 2020, Chinese Patent Application No. 202010947590.4, filed on September 10, 2020, and Chinese Patent Application No. 202110587528.3, filed on May 27, 2021, which are incorporated herein by reference in their entireties.

[0002] The present invention relates to the field of medicinal chemistry, in particular to pyrazine compounds, methods for their preparation and the use of the compounds as EGFR inhibitors in the preparation of antitumor drugs. [Background technology]

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

[0004] The EGFR family is a group of protein kinases involved in the transmission of mitogenic signals and plays an important role in growth and development. Extensive analyses and studies of in vitro tumor cells, animal models, and human tumor samples have shown that mutations in EGFR family members may drive human tumor progression and be one of the key causes of the development and progression of many cancers. Therefore, targeting EGFR mutant proteins and inhibiting their activity is an important means for treating related tumors.

[0005] Studies have shown that EGFR gene mutations can be found in approximately 12% to 47% of non-small cell lung cancer (NSCLC). The two most common EGFR gene mutations in NSCLC are exon 19 deletion (del19) and the L858R missense mutation in exon 21. These mutations result in persistent activation of the EGFR protein, independent of ligand. NSCLC patients with EGFR del19 or L858R mutations are more sensitive to targeted therapy with EGFR protein kinase inhibitors (EGFR TKIs), such as erlotinib, gefitinib, afatinib, or osimertinib, and have demonstrated high objective response rates (ORRs) in clinical trials (approximately 60%–85%). However, these responses usually do not last long enough, and most patients treated with first- or second-generation EGFR TKIs experience disease progression within approximately 11 months. Analysis of drug resistance has shown that in approximately 50% to 70% of drug-resistant patients, the molecular mechanism of drug resistance is the acquisition of a secondary mutation, the T790M mutation (T790M+), in the EGFR gene (Cancer discov. 2012, 2:872-5). This secondary mutation causes the loss of inhibitory activity of first- and second-generation EGFR TKIs against mutant tumor cells.

[0006] Osimertinib, a third-generation covalent EGFR TKI, is being developed for the treatment of tumors harboring EGFR del19 and L858R mutations, regardless of the presence or absence of the T790M mutation. Osimertinib demonstrates high response rates despite drug resistance induced by the T790M mutation; however, approximately 70% of patients eventually develop drug resistance and progress after approximately 10 months (Lung Cancer. 2017, 108:228-231). Research into the molecular mechanisms of drug resistance to third-generation EGFR TKIs has revealed that in approximately 20%–40% of patients who relapse after receiving osimertinib, the acquisition of a third mutation in the EGFR gene, the C797S mutation, is one of the primary mechanisms of drug resistance. Furthermore, patients with EGFR del19 / L858R T790M C797S mutations become refractory to first-, second-, and third-generation EGFR TKIs after treatment with a third-generation EGFR TKI. In 2015, Thress et al. reported the first analysis of resistance to osimertinib in 15 patients, revealing that approximately 40% of drug resistance cases were due to the C797S mutation (Nature Medicine, 2015, 21:560-562). At the 2017 American Society of Clinical Oncology (ASCO) Annual Meeting, Piotrowska and Caicun Zhou reported analyses of drug resistance in 23 and 99 cases, respectively, demonstrating that approximately 22% of drug resistance cases were due to the C797S mutation. Therefore, targeting and inhibiting the EGFR del19 / L858R T790M C797S mutation could overcome resistance to osimertinib. However, there are currently no commercially available EGFR TKIs capable of inhibiting the EGFR del19 / L858R T790M C797S mutation, necessitating the urgent research and discovery of fourth-generation EGFR TKIs to address this clinical need.

[0007] The EGFR del19 / L858R T790M C797S mutation is a new EGFR mutation that arose after treatment with third-generation EGFR TKIs and has not been fully interpreted. Currently, only a few fourth-generation EGFR TKIs that inhibit the EGFR del19 / L858R T790M C797S mutation have been reported. For example, Boehringer Ingelheim has reported a class of macrocyclic compounds, BI-4020, that have anti-EGFR del19 / L858R T790MC797S mutant activity and in vivo antitumor activity (J. Med. Chem. 2019, 62:10272-10293). International Publication No. 2019 / 015655 reported a class of aryl-phosphorus-oxy compounds that have anti-EGFR del19 / L858R T790M C797S mutant activity and in vivo antitumor activity. General formula A and its representative compound B (Example 41) are shown in the following structures (see the present invention for the definition of the symbols in the formula). [ka] [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication No. 2019 / 015655 [Non-patent literature]

[0009] [Non-Patent Document 1] Cancer discov.2012,2:872-5 [Non-patent document 2] Nature Medicine,2015,21:560-562 [Non-patent document 3] J.Med.Chem.2019,62:10272-10293 Summary of the Invention [Problem to be solved by the invention]

[0010] Currently, there is an urgent need to search for and discover compounds that have good activity against the EGFR del19 / L858R T790M C797S mutant. [Means for solving the problem]

[0011] Summary of the Invention An object of the present invention is to provide a compound of general formula (1), or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate, or solvate thereof. [ka] Here, in general formula (1), Y is a 3- to 11-membered heterocycloalkyl, a C6 to C14 aryl, or a 5- to 10-membered heteroaryl, wherein the heterocycloalkyl, the aryl, and the heteroaryl are each independently selected from the group consisting of —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)NR 4 R 5 and —O—CH—O—, L 1 is -O- or -NH-, X is a C6-C14 arylene or a 5- to 11-membered heteroarylene, and the arylene and heteroarylene are optionally substituted with one or more groups selected from the group consisting of -H, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, and C1-C6 haloalkoxy; R 1 is -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 to 15-membered heterocycloalkyl or 3 to 15-membered heterocycloalkyl, wherein the alkoxy and the heterocycloalkyl are each independently —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 and optionally substituted with one or more groups of L 2 is -O-, -NH- or a chemical bond, R 2 represents C1-C6 alkyl, C3-C14 cycloalkyl, C6-C14 aryl, 3- to 4-membered heterocycloalkyl, [ka] or a 6- to 11-membered heterocycloalkyl, wherein the alkyl, the cycloalkyl, the aryl, the heterocycloalkyl, [ka] -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)NR 4 R 5 and optionally substituted with one or more groups of R 3 is a 3- to 11-membered heterocycloalkyl, wherein the heterocycloalkyl is selected from the group consisting of -H, -CD3, -R 4 , -OR 4 and -NR 4 R 5 and optionally substituted with one or more groups of R 4 and R 5 are each independently —H, C1-C6 alkyl, or C3-C14 cycloalkyl; R 6 and R 7 are each independently —H, C1-C6 alkyl, or C3-C14 cycloalkyl, or R 6 and R 7 together with the N atom attached thereto form a 3- to 11-membered heterocycloalkyl, wherein said heterocycloalkyl is selected from the group consisting of -H, -CD 3 , halogen, -R 4 AND-OR 4 and optionally substituted with one or more groups of R 0 is C1-C6 alkyl or C3-C14 cycloalkyl, 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.

[0012] In another preferred embodiment, in general formula (1), Y is a 5- to 6-membered heterocycloalkyl, phenyl, or 5- to 9-membered heteroaryl, wherein the heterocycloalkyl, the phenyl, and the heteroaryl may be optionally substituted with one or more groups selected from the group consisting of -H, -F, -Cl, -Br, -CN, -OH, -OCH3, -NH2, -N(CH3)2, -NHCOCH3, -NHSO2CH3, -CH3, -CONH2, -CH2OH, and -O-CH2-O-.

[0013] In another preferred embodiment, in general formula (1), Y is [ka] [ka] is.

[0014] In another preferred embodiment, in the general formula (1), X is phenylene or 6-membered heteroarylene, wherein the phenylene and the heteroarylene are -H, -F, -CH3, -CH2CH3, -CH(CH3)2, [ka] , -OCH3, -OCF2H and -OCF3.

[0015] In another preferred embodiment, in general formula (1), X is [ka] is.

[0016] In another preferred embodiment, in the general formula (1), R1 is -H, -N(CH3)2, -CH2-6 to 11-membered heterocycloalkyl or 6 to 11-membered heterocycloalkyl, wherein the heterocycloalkyl is [ka] TIFF0007777544000010.tif30151, -H, -CH3, [ka] , -N(CH3)2, [ka] and -CD3.

[0017] In another preferred embodiment, in general formula (1), R1 is -H, -N(CH3) 2、 [ka] is.

[0018] In another preferred embodiment, in general formula (1), L 2 When is -NH-, R 2 teeth, [ka] is.

[0019] In another preferred embodiment, in general formula (1), L 2 When is -O-, R 2 teeth, [ka] is.

[0020] In another preferred embodiment, in general formula (1), L 2 When is a chemical bond, R 2 teeth, [ka] is.

[0021] In various embodiments, representative compounds of the present invention have the following structure: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] It has one of the following.

[0022] The present invention further contemplates providing a pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier and a compound of general formula (1) disclosed herein or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient.

[0023] The present invention further contemplates the use of a compound disclosed herein, or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof, or said pharmaceutical composition, in preparing a medicament for treating a disease associated with an EGFR mutation.

[0024] The present invention further provides a method for treating, regulating and / or preventing a disease associated with an EGFR mutant protein, comprising the step of administering a therapeutically effective amount of the compound, or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof, or the pharmaceutical composition to a subject. Through the synthesis and careful study of various novel compounds having EGFR inhibitory activity, the present inventors have surprisingly found that the compound of general formula (1) inhibits EGFR. del19 / T790M / C797S and EGFRL 858R / T790M / C797S and when Y is heterocycloalkyl, aromatic heterocycle, or aryl, it has high selectivity against wild-type EGFR WT.

[0025] It is to be understood that both the foregoing general description and the following detailed description of the invention are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 1 shows the results of tumor growth inhibition in an in vivo pharmacodynamic test using mice according to Example 4 of the present invention. [Figure 2] FIG. 1 shows the results of tumor growth inhibition in an in vivo pharmacodynamic test using mice according to Example 5 of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0027] Compound synthesis Methods for preparing the compounds of the general formula (1) of the present invention are specifically described below, but these specific methods do not limit the present invention.

[0028] The compounds of formula (1) above can be synthesized using standard synthetic techniques, well-known techniques, in combination with the methods described herein. Furthermore, the solvents, temperatures, and other reaction conditions described herein may vary. Starting materials for the synthesis of the compounds can be obtained synthetically or commercially. The compounds described herein and other related compounds with different substituents are described 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), and Green and Wuts, Protective Groups in Organic Synthesis, 3 rd Ed., (Wiley 1999). General methods for preparing compounds can be modified by using appropriate reagents and conditions to introduce different groups into the formulas provided herein.

[0029] In one embodiment, the compounds described herein are prepared according to methods well known in the art. However, the conditions of the methods, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to the following explanations. In addition, the compounds of the present invention can be easily prepared by any combination of various synthetic methods described herein or known in the art, and such combinations can be easily determined by those skilled in the art to which the present invention pertains. In one embodiment, the present invention also provides a method for preparing a compound of general formula (1) prepared using the following general reaction scheme 1 or general reaction scheme 2.

[0030] General Reaction Scheme 1 [ka]

[0031] Embodiments of compounds of general formula (1) can be prepared according to General Reaction Scheme 1, where R 1 , R 2 , X, Y, L 1 and L 2 is as defined above, H represents hydrogen, and B represents boric acid, borate, or trifluoroborate. As shown in General Reaction Scheme 1, compound 1-1 is reacted with formamide to give compound 1-2, and compound 1-2 can be prepared by reacting R 1 -XL 1 Compound 1-3 is obtained by reacting compound 1-3 with YB to give compound 1-4, and compound 1-4 is reacted with R 2 -L 2 Reaction with —H affords the target compounds 1-5.

[0032] General Reaction Scheme 2 [ka]

[0033] Embodiments of compounds of general formula (1) can be prepared according to General Reaction Scheme 2, where R 1 , R2 , X, Y, L 1 and L 2 is as defined above, and H represents hydrogen. As shown in General Reaction Scheme 2, compound 2-1 is reacted with formamide to give compound 2-2, which can be prepared by reacting compound 2-2 with R 1 -XL 1 -H to give compound 2-3, which is then reacted with R 2 -L 2 -H to give compound 2-4, which is then reacted with YH under basic conditions to give the target compound 2-5.

[0034] Further forms of the compound As used herein, "pharmaceutically acceptable" refers to a substance, such as a carrier or diluent, that does not cause a compound to lose its biological activity or properties, that is relatively non-toxic, e.g., that does not cause undesired biological effects or adverse interactions with any of its components when the substance is administered to an individual.

[0035] The term " pharmaceutically acceptable salt " refers to the form of a compound that does not cause significant stimulation to organisms due to drug administration or does not eliminate the biological activity and properties of the compound.In certain embodiments, pharmaceutically acceptable salts can be obtained by reacting the compound of general formula (1) with an acid, for example, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid and nitric acid, organic acids such as 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, acidic amino acids such as aspartic acid and glutamic acid.

[0036] It should be understood that pharmaceutically acceptable salts include solvent addition forms or crystalline forms, particularly solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and are selectively formed upon crystallization with pharmaceutically acceptable solvents such as water and ethanol. Hydrates are formed when the solvent is water, and alcoholates are formed when the solvent is ethanol. Solvates of compounds of general formula (1) are conveniently prepared or formed according to the methods described herein. For example, hydrates of compounds of general formula (1) are conveniently prepared by recrystallization from a water / organic solvent mixture, where the organic solvent used includes, but is not limited to, tetrahydrofuran, acetone, ethanol, or methanol. Furthermore, the compounds referred to herein can exist in both unsolvated and solvated forms. In general, solvated forms are considered equivalent to unsolvated forms for the purposes of the compounds and methods provided herein.

[0037] In other specific examples, compounds of general formula (1) are prepared in different forms, including, but not limited to, amorphous, crushed, and nanoparticle forms. Furthermore, compounds of formula (1) may include crystalline forms and may also be polymorphs. Polymorphs contain different lattice arrangements of the same elements of a compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystalline forms, optical properties, electrical properties, stability, and solubility. Different factors, such as recrystallization solvents, crystallization rates, and storage temperatures, may result in the predominance of a single crystal.

[0038] In another embodiment, the compounds of general formula (1) may have chiral centers and / or axial chirality and therefore may exist in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds, single diastereomers, and cis-trans isomers. Each chiral center or axial chirality independently produces two optical isomers, and all possible optical isomers, diastereomeric mixtures, and pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomers of these compounds.

[0039] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be labeled with radioactive isotopes such as tritium (H), iodine-125 (I), and C-14 (C). As another example, deuterium can be used in place of hydrogen atoms to form deuterated compounds. The bond formed between deuterium and carbon is stronger than the bond formed between normal hydrogen and carbon. Compared to non-deuterated pharmaceuticals, deuterated pharmaceuticals generally have advantages such as reduced toxicity and side effects, increased pharmaceutical stability, improved therapeutic effects, and prolonged in vivo half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

[0040] Terminology Unless otherwise indicated, terms used in the specification and claims are defined as follows: In this specification and the appended claims, the singular forms "a" and "an" include the plural reference unless the context clearly dictates otherwise. Conventional methods such as mass spectrometry, nuclear magnetic resonance spectroscopy, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are employed unless otherwise indicated. As used herein, "or" means "and / or" unless otherwise indicated.

[0041] Unless otherwise specified, "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight-chain and branched groups, having 1 to 14 carbon atoms. Lower alkyls having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, or tert-butyl, are preferred. As used herein, "alkyl" includes unsubstituted and substituted alkyls, particularly alkyls substituted with one or more halogens. Preferred alkyls are CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, i Pr, n Pr, iBu, n Bu and t Bu.

[0042] Unless otherwise specified, "alkenyl" refers to an unsaturated aliphatic hydrocarbon group having a carbon-carbon double bond, including straight-chain or branched groups containing 1 to 14 carbon atoms. Lower alkenyl groups containing 1 to 4 carbon atoms, such as vinyl, 1-propenyl, 1-butenyl, or 2-methylpropenyl, are preferred.

[0043] Unless otherwise specified, "alkynyl" refers to an unsaturated aliphatic hydrocarbon group having a carbon-carbon triple bond, including straight-chain and branched groups containing 1 to 14 carbon atoms. Lower alkynyl groups containing 1 to 4 carbon atoms, such as ethynyl, 1-propynyl, or 1-butynyl, are preferred.

[0044] Unless otherwise specified, "cycloalkyl" refers to a 3- to 14-membered all-carbon monocyclic aliphatic hydrocarbon group in which one or more rings may contain one or more double bonds, but none has a completely conjugated pi-electron system, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexane, and cyclohexadiene.

[0045] Unless otherwise specified, "alkoxy" refers to an alkyl group attached to the remainder of the molecule via an ether oxygen atom. Representative alkoxy groups are those having 1 to 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, particularly alkoxy substituted with one or more halogens. Preferred alkoxy are OCH3, OCF3, CHF2O, CF3CHO, i- PrO, n- PrO, i- BuO, n- BuO and t- BuO.

[0046] Unless otherwise specified, "aryl" refers to a monocyclic or polycyclic aromatic hydrocarbon group. For example, a monocyclic aryl ring may be fused with one or more carbocyclic aromatic groups. Examples of aryl include, but are not limited to, phenyl, naphthyl, and phenanthryl.

[0047] Unless otherwise specified, "arylene" refers to a divalent aryl as defined above. Examples of arylene include, but are not limited to, phenylene, naphthylene, phenanthrylene, and the like.

[0048] Unless otherwise specified, "heteroaryl" refers to a monocyclic or polycyclic aromatic group containing one or more heteroatoms (O, S, or N); for example, a monocyclic heteroaryl ring can be fused with one or more carbocyclic aromatic groups or other monocyclic heterocyclyl groups. Examples of heteroaryl include, but are not limited to, pyridinyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, benzopyridyl, and pyrrolopyrimidinyl.

[0049] Unless otherwise specified, "heteroarylene" refers to a divalent heteroaryl as defined above.

[0050] Unless otherwise specified, "heterocycloalkyl" refers to a saturated or partially unsaturated ring system radical 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 specified, a "heterocycloalkyl" ring system can be a monocyclic, bicyclic, spirocyclic, or polycyclic ring system. A "heterocycloalkyl" can be attached to the remainder of the molecule through one or more ring carbons or heteroatoms. Examples of "heterocycloalkyl" include, but are not limited to, pyrrolidine, piperidine, N-methylpiperidine, tetrahydroimidazole, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, pyrimidine-2,4(1H,3H)-dione, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, 2-azaspiro[3.3]heptane, and the like.

[0051] Unless otherwise specified, "halogen" (or halo) refers to fluorine, chlorine, bromine, or iodine. The term "halo" (or "halogenated") before a group name indicates that the group is partially or fully halogenated, i.e., substituted by F, Cl, Br, or I, preferably F or Cl, in any combination.

[0052] "Optional" or "optionally" means that the subsequently described event or circumstance may occur, but does not necessarily occur, and the description includes instances in which the event or circumstance occurs and instances in which it does not occur.

[0053] The substituent "-O-CH-O-" means that the two oxygen atoms in the substituent are attached to two adjacent carbon atoms in a heterocycloalkyl, aryl, or heteroaryl, for example: [ka]

[0054] When the number of a linker group is 0, such as -(CH2)0-, it means that the linker group is a single bond.

[0055] When one of the variables is selected from a chemical bond, it means that the two groups linked by this variable are directly linked. For example, when L in XLY represents a chemical bond, it actually means that the structure is XY.

[0056] Specific pharmaceutical and medical terms As used herein, the term "acceptable" means that the formulation or active ingredient does not have excessively deleterious effects on the health of the general subject being treated.

[0057] As used herein, the terms "treatment," "course of treatment," or "treatment" include alleviating, suppressing, or ameliorating a disease symptom or condition, inhibiting the development of a complication, improving or preventing underlying metabolic syndrome, inhibiting the development of a disease or condition (e.g., controlling the progression of a disease or condition), alleviating a disease or symptom, relieving a disease or symptom, alleviating a complication caused by a disease or condition, or preventing or treating a symptom caused by a disease or condition. As used herein, a compound or pharmaceutical composition, when administered, can ameliorate a disease, symptom, or condition, and in particular, can reduce the severity, delay the onset, slow the progression, or shorten the duration of a disease. Fixed or temporary administration, or continuous or intermittent administration, can result from or relate to administration.

[0058] The term "active ingredient" refers to compounds of formulas (1) to (3) and pharmaceutically acceptable inorganic or organic salts of compounds of formulas (1) to (3). The compounds of the present invention may contain one or more asymmetric centers (axial chirality) and therefore may exist in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds, and single diastereomers. The possible asymmetric centers depend on the properties of the various substituents on the molecule. Each of these asymmetric centers independently generates two optical isomers, and all possible optical isomers, diastereomeric mixtures, and pure or partially pure compounds are within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

[0059] As used herein, terms such as "compound," "composition," "drug," or "medicine or pharmaceutical agent" are used interchangeably and refer to any compound or composition that, when administered to an individual (human or animal), is capable of eliciting a desired pharmacological and / or physiological response through local and / or systemic action.

[0060] The terms "administered, administering, or administration" as used herein refer to direct administration of a compound or composition, or the administration of a prodrug, derivative, analog, etc. of an active compound.

[0061] While the numerical ranges and parameters defining the broad scope of the present invention are approximations, the relevant values ​​set forth in specific embodiments are set forth herein as precisely as possible. However, any numerical value inherently contains standard deviations necessarily resulting from certain testing methods. Herein, the term "about" generally means that the actual value is within ±10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "about" indicates that the actual value falls within an acceptable standard error of the mean, as would be expected by one of ordinary skill in the art. Except in experimental examples or unless otherwise indicated, all ranges, amounts, values, and percentages used herein (e.g., to describe amounts of materials, lengths of time, temperatures, operating conditions, proportions of amounts, etc.) are understood to be modified by the term "about." Accordingly, unless otherwise indicated, all numerical parameters set forth in this specification and the appended claims are approximations that may vary, if desired. At the very least, these numerical parameters should be construed as representing significant digits or as derived using conventional rounding rules.

[0062] Scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art, unless otherwise defined herein. Furthermore, as used herein, singular nouns include their plural forms, and plural nouns as used herein also include their singular forms, unless contradictory to the context.

[0063] therapeutic use The present invention provides methods of treating diseases, including but not limited to diseases associated with EGFR mutations (eg, cancer), using the compounds or pharmaceutical compositions disclosed herein.

[0064] In some embodiments, a method for treating cancer is provided, comprising administering to an individual in need thereof an effective amount of any of the aforementioned pharmaceutical compositions comprising a compound of structural formula (1). 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 cancer, prostate cancer, ovarian cancer, head and neck cancer, gastric cancer, mesothelioma, or all cancer metastases.

[0065] Administration route The compounds of the present invention and their pharmaceutically acceptable salts can be prepared into various formulations containing the compounds disclosed herein or their pharmaceutically acceptable salts and pharmaceutically acceptable excipients or carriers within a safe and effective amount, where "safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount of the compound is determined according to the age, condition, course of treatment, and other specific conditions of the subject to be treated.

[0066] A "pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gel substances that must be suitable for human use and have sufficient purity and low toxicity. "Compatible" here means that the components of the composition can be intermixed with the compounds of the present invention without significantly reducing the pharmaceutical efficacy of the compounds. Examples of pharmaceutically acceptable excipients or carriers include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, or cellulose acetate), gelatin, talc, solid lubricants (e.g., stearic acid or magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween®), wetting agents (e.g., sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0067] The compounds of the present invention can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously) or topically.

[0068] Solid dosage forms for oral administration include capsules, tablets, pills, pulvises, 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 the following ingredients: (a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants such as glycerol; (d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retardants such as paraffin; (f) absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glycerol monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, and sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, said dosage forms may also comprise buffering agents.

[0069] Solid dosage forms such as tablets, sugar-coated tablets, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other materials known in the art.These may contain opacifying agents, and the active compound or compounds in such compositions can be delayed in certain parts of the digestive tract.The examples of embedding components that can be used include polymeric materials and wax-based materials.If necessary, the active compound can be formed into microcapsules with one or more of the above-mentioned excipients.

[0070] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, etc. In addition to the active compound, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents 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.

[0071] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0072] Suspensions may contain, in addition to the active compound, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methylate, and agar-agar, or mixtures of these substances.

[0073] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or non-aqueous 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.

[0074] Dosage forms for topical administration of the compounds of the present invention include ointments, pulps, patches, sprays, inhalants, etc. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier, and any preservatives, buffers, or propellants that may be required as needed.

[0075] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0076] When the pharmaceutical composition of the present invention is used, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., human) to be treated, and the dosage is a pharmaceutically effective dosage. For a human weighing 60 kg, the daily dosage is usually 1 to 2,000 mg, preferably 50 to 1,000 mg. The specific dosage is determined taking into consideration factors such as the route of administration and the patient's health condition, which are well known to those skilled in the art.

[0077] The above-described features of the present invention or the features described in the embodiments can be arbitrarily combined. All features disclosed herein can be used in any composition, and various features disclosed herein can be replaced with any alternative features that serve the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely generic examples of equivalent or similar features.

[0078] (Detailed explanation) Various specific aspects, features and advantages of the above compounds, methods and pharmaceutical compositions are described in detail below, thereby clarifying the present invention. It should be understood that the following detailed description and examples describe specific embodiments for reference. After reading the description of the present invention, those skilled in the art may make various changes or modifications to the present invention, and such equivalents also fall within the scope of the present invention as defined herein.

[0079] In all examples, 1H-NMR spectra were recorded on a Vian Mercury 400 nuclear magnetic resonance spectrometer, and chemical shifts were expressed in δ (ppm). Unless otherwise specified, 200-300 mesh silica gel was used for separation, and the ratio of eluents was by volume.

[0080] In the present invention, the following abbreviations are used: CDCl3 is deuterated chloroform; CD3OD is deuterated methanol; DMSO-d6 is deuterated dimethyl sulfoxide; EtOAc is ethyl acetate; hexane is n-hexane; MeCN is acetonitrile; DCM is dichloromethane; DIPEA is diisopropylethylamine; NMP is 1-methylpyrrolidin-2-one; dioxane is 1,4-dioxane; DMF is N,N-dimethylformamide; DMSO is dimethyl sulfoxide; h is hour; K3PO4 is potassium phosphate; min: minute, MS: mass spectrometry, NaH: sodium hydride, NMR: nuclear magnetic resonance, Pd2(dba)3: tris(dibenzylideneacetone)dipalladium, Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, TFA (CF3COOH): trifluoroacetic acid, TLC: thin-layer chromatography, THF: tetrahydrofuran, and Xantphos: 4,5-bis(diphenylphosphane)-9,9-dimethylxanthene.

[0081] Synthesis method A: Synthesis of Compound 135 (5-((3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide) and its optical isomers (Compounds 136, 137, 138, and 139) using Synthetic Method A

[0082] [ka]

[0083] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2) 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-neck flask, and the mixture was heated to 90 °C with stirring. Solid (NH)SO (25 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 2 h. Solid KSO (30 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 20 h. The reaction was monitored by LC-MS, showing remaining starting material. EtOAc (150 mL) and water (300 mL) were added to the mixture, stirred, and separated. The aqueous phase was again extracted with EtOAc (150 mL). The combined organic phase was washed with saturated aqueous sodium chloride (150 mL), concentrated, and the residue was purified by column chromatography (Et0Ac:Hexane = 0:1 to 1:5 to 1:2) to give the product (1.82 g, 10.5% yield). The remaining starting material was recovered (10.3 g, 68.7% yield). 1 H NMR (400 MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H]+.

[0084] 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): 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 one-neck flask. The mixture was purged with argon, stirred, and heated at reflux for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was concentrated, and the residue was purified by column chromatography to give the product (368 mg, 75% yield). 1H NMR (400 MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J = 3.8 Hz, 1H), 7.51-7.44 (m, 2H), 6.99-6.88 (m, 2H), 5.67 (d, J = 3.9 Hz, 1H), 3.80-3.63 (m, 2H), 2.84-2.42 (m, 10H), 2.39 (ddt, J = 11.4, 7.3, 3.7 Hz, 1H), 1.96 (dt, J = 12.2, 3.0 Hz, 2H), 1.70 (qd, J = 12.1, 4.0 Hz, 2H); MS (ESI): 556 [M+H] + .

[0085] 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): 5-Chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (167 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), phenylboronic acid (40.23 mg, 0.33 mmol), dioxane / HO (10 mL / 2 mL), and Pd(dppf)Cl (22 mg) were added to a 50 mL single-neck flask. The mixture was purged with argon, rapidly heated to 105 °C, and incubated for 30 min. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (115 mg, 75.6% yield). 1H NMR (400 MHz, CDCl3) δ: 10.74 (s, 1H), 7.70 (d, J = 6.5 Hz, 3H), 7.56 (d, J = 8.5 Hz, 2H), 7.44 (p, J = 6.8 Hz, 3H), 6.94 (d, J = 8.6 Hz, 2H), 5.67 (s, 1H), 3.70 (d, J = 11.9 Hz, 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 = 12.4 Hz, 2H), 1.67 (tt, J= 12.5, 6.8 Hz, 2H); LC-MS: 506 [M+H] + .

[0086] 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): 5-Chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (133.6 mg, 0.27 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (31 mg, 0.54 mmol), DMSO (5 mL), and 4 Å molecular sieves (200 mg, powder) were added to a 50 mL one-neck flask. The mixture was purged with argon and stirred at room temperature for 15 minutes. Next, 3-aminocyclopentanol hydrochloride (45 mg, 0.32 mmol) was added, and the mixture was purged with argon, heated to 120 °C, and stirred for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (92 mg, 59.7% yield). MS (ESI): 571 [M+H] + .

[0087] Chiral separation afforded four optically pure chiral isomers. [ka]

[0088] 5-(((1R,3S)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 136): 1 H NMR (400 MHz, 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 = 6.5 Hz, 1H), 5.13 (s, 1H), 4.56 (h, J= 7.1 Hz, 1H), 4.39 (tt, J = 5.8, 2.9 Hz, 1H), 3.69 (d, J = 12.0 Hz, 2H), 2.80-2.58 (m, 5H), 2.50 (s, 3H), 2.35 (d, J = 9.8 Hz, 1H), 2.30 (s, 3H), 2.29-2.22 (m, 1H), 2.18 (dd, J = 13.8, 7.2 Hz, 1H), 2.06-1.96 (m, 1H), 1.92 (d, J = 12.4 Hz, 2H), 1.71-1.54 (m, 6H), 1.43 (ddd, J= 13.0, 9.1, 6.6 Hz, 1H); MS (ESI): 571 [M+H] + .

[0089] 5-(((1R,3R)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 137): 1H NMR (400 MHz, CDCl3) δ: 10.78 (s, 1H), 7.60 (dd, J = 8.6, 6.7 Hz, 4H), 7.44 (t, J = 7.5 Hz, 3H), 7.36 (t, J = 7.3 Hz, 1H), 6.91 (d, J = 8.9 Hz, 2H), 6.04 (d, J = 7.4 Hz, 1H), 5.10 (s, 1H), 4.54 (d, J = 7.8 Hz, 1H), 4.44 (s, 1H), 3.68 (d, J = 11.9 Hz, 2H), 2.77-2.57 (m, 5H), 2.48 (s, 3H), 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] + .

[0090] 5-(((1S,3S)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 138): 1H NMR (400 MHz, 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 = 6.5 Hz, 1H), 5.13 (s, 1H), 4.56 (h, J= 7.1 Hz, 1H), 4.39 (tt, J = 5.8, 2.9 Hz, 1H), 3.69 (d, J = 12.0 Hz, 2H), 2.80-2.58 (m, 5H), 2.50 (s, 3H), 2.35 (d, J = 9.8 Hz, 1H), 2.30 (s, 3H), 2.29-2.22 (m, 1H), 2.18 (dd, J = 13.8, 7.2 Hz, 1H), 2.06-1.96 (m, 1H), 1.92 (d, J = 12.4 Hz, 2H), 1.71-1.54 (m, 6H), 1.43 (ddd, J= 13.0, 9.1, 6.6 Hz, 1H); MS (ESI): 571 [M+H] + .

[0091] 5-(((1S,3R)-3-hydroxycyclopentyl)amino)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (compound 139): 1H NMR (400 MHz, CDCl3) δ: 10.78 (s, 1H), 7.60 (dd, J = 8.6, 6.7 Hz, 4H), 7.44 (t, J = 7.5 Hz, 3H), 7.36 (t, J = 7.3 Hz, 1H), 6.91 (d, J = 8.9 Hz, 2H), 6.04 (d, J = 7.4 Hz, 1H), 5.10 (s, 1H), 4.54 (d, J = 7.8 Hz, 1H), 4.44 (s, 1H), 3.68 (d, J = 11.9 Hz, 2H), 2.77-2.57 (m, 5H), 2.48 (s, 3H), 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] + .

[0092] Synthesis of 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)pyrazine-2-carboxamide) using Synthesis Method A [ka]

[0093] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2) 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-neck flask, and the mixture was heated to 90 °C with stirring. Solid (NH)SO (25 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 2 h. Solid KSO (30 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 20 h. The reaction was monitored by LC-MS, showing remaining starting material. EtOAc (150 mL) and water (300 mL) were added to the mixture, stirred, and separated. The aqueous phase was again extracted with EtOAc (150 mL). The combined organic phase was washed with saturated aqueous sodium chloride (150 mL), concentrated, and the residue was purified by column chromatography (Et0Ac:Hexane = 0:1 to 1:5 to 1:2) to give the product (1.82 g, 10.5% yield). The remaining starting material was recovered (10.3 g, 68.7% yield). 1 H NMR (400 MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0094] 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): 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 one-neck flask. The mixture was purged with argon, stirred, and heated at reflux for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was concentrated, and the residue was purified by column chromatography to give the product (368 mg, 75% yield). 1H NMR (400 MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J = 3.8 Hz, 1H), 7.51-7.44 (m, 2H), 6.99- 6.88 (m, 2H), 5.67 (d, J = 3.9 Hz, 1H), 3.80-3.63 (m, 2H), 2.84-2.42 (m, 10H), 2.39 (ddt, J = 11.4, 7.3, 3.7 Hz, 1H), 1.96 (dt, J = 12.2, 3.0 Hz, 2H), 1.70 (qd, J = 12.1, 4.0 Hz, 2H); MS (ESI): 556 [M+H] + .

[0095] Step 3: Synthesis of compound 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(pyridin-4-yl)pyrazine-2-carboxamide (compound int_6) 5-Chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (166.76 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), (pyridin-4-yl)boronic acid (40.56 mg, 0.33 mmol), dioxane / HO (10 mL / 2 mL), and Pd(dppf)Cl (22 mg) were added to a 50 mL single-neck flask. The mixture was purged with argon, rapidly heated to 105 °C, and incubated for 60 min. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (126 mg, 82.8% yield). MS (ESI): 507 [M+H]+.

[0096] 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)pyrazine-2-carboxamide (compound 39). 5-Chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(pyridin-4-yl)pyrazine-2-carboxamide (152.1 mg, 0.3 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (35 mg, 0.6 mmol), DMSO (5 mL), and 4 Å molecular sieves (200 mg, powder) were added to a 50 mL single-neck flask. The mixture was purged with argon and stirred at room temperature for 15 minutes. Next, 3-tetrahydro-2H-pyran-4-amine (33.4 mg, 0.33 mmol) was added, and the mixture was purged with argon, heated to 120 °C, and stirred for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (130 mg, 75.8% yield). 1 H NMR (400 MHz, CDCl3) δ 10.86 (s, 1H), 8.71 (d, J = 5.1 Hz, 2H), 7.55 (t, J = 6.8 Hz, 4H), 7.42 (s, 1H), 6.91 (d, J = 8.8 Hz, 2H), 5.22 (s, 1H), 5.14 (d, J= 7.0 Hz, 1H), 4.17 (m, 1H), 4.01 (d, J = 11.7 Hz, 2H), 3.69 (d, J= 9.2 Hz, 2H), 3.52 (t, J = 11.6 Hz, 2H), 2.69 (m, 11H), 2.38 (s, 3H), 2.09-1.96 (m, 4H), 1.76-1.64 (m, 4H); MS (ESI): 572 [M+H] + .

[0097] Synthesis of 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)pyrazine-2-carboxamide) using Synthesis Method A [ka]

[0098] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2): 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-neck flask, and the mixture was heated to 90 °C with stirring. Solid (NH)SO (25 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 2 h. Solid KSO (30 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 20 h. The reaction was monitored by LC-MS, showing remaining starting material. EtOAc (150 mL) and water (300 mL) were added to the mixture, stirred, and separated. The aqueous phase was again extracted with EtOAc (150 mL). The combined organic phase was washed with saturated aqueous sodium chloride (150 mL), concentrated, and the residue was purified by column chromatography (Et0Ac:Hexane = 0:1 to 1:5 to 1:2) to give the product (1.82 g, 10.5% yield). The remaining starting material was recovered (10.3 g, 68.7% yield). 1 H NMR (400 MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0099] 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): 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 one-neck flask. The mixture was purged with argon, stirred, and heated at reflux for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was concentrated, and the residue was purified by column chromatography to give the product (368 mg, 75% yield). 1 H NMR (400 MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J = 3.8 Hz, 1H), 7.51-7.44 (m, 2H), 6.99- 6.88 (m, 2H), 5.67 (d, J = 3.9 Hz, 1H), 3.80-3.63 (m, 2H), 2.84-2.42 (m, 10H), 2.39 (ddt, J = 11.4, 7.3, 3.7 Hz, 1H), 1.96 (dt, J = 12.2, 3.0 Hz, 2H), 1.70 (qd, J = 12.1, 4.0 Hz, 2H); MS (ESI): 556 [M+H] + .

[0100] Step 3: Synthesis of compound 5-chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(1H-pyrazol-3-yl)pyrazine-2-carboxamide (compound int_7) 5-Chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (166.76 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), (1H-pyrazol-3-yl)boronic acid (37 mg, 0.33 mmol), dioxane / HO (10 mL / 2 mL), and Pd(dppf)Cl (22 mg) were added to a 50 mL single-neck flask. The mixture was purged with argon, rapidly heated to 105 °C, and incubated for 60 min. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (119 mg, 80% yield). MS (ESI): 496 [M+H] + .

[0101] 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)pyrazine-2-carboxamide (compound 55): 5-Chloro-3-((4-4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-(1H-pyrazol-3-yl)pyrazine-2-carboxamide (148.8 mg, 0.3 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (35 mg, 0.6 mmol), DMSO (5 mL), and 4 Å molecular sieves (200 mg, powder) were added to a 50 mL single-neck flask. The mixture was purged with argon and stirred at room temperature for 15 minutes. Next, 3-tetrahydro-2H-pyran-4-amine (33.4 mg, 0.33 mmol) was added, and the mixture was purged with argon, heated to 120 °C, and stirred for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (115 mg, 68.3% yield). Compound 55 fumarate: 1 H NMR (400 MHz, DMSO-d6) δ 13.05 (s, 1H), 11.21 (s, 1H), 8.90 (s, 1H), 8.00-7.92 (m, 1H), 7.81 (d, J = 2.4 Hz, 1H), 7.55-7.47 (m, 2H), 7.31 (d, J = 2.8 Hz, 1H), 7.21 (d, J = 2.4 Hz, 1H), 6.89 (d, J = 8.9 Hz, 2H), 6.54 (s, 4H), 4.11 (m, J = 6.2 Hz, 1H), 3.89 (dt, J= 11.5, 3.7 Hz, 2H), 3.62 (d, J = 11.8 Hz, 2H), 3.49 (td, J = 11.5, 2.4 Hz, 2H), 2.88-2.52 (m, 9H), 2.42 (s, 3H), 2.10-1.99 (m, 2H), 1.84 (d, J = 11.1 Hz, 2H), 1.61-1.40 (m, 4H); MS (ESI): 561 [M+H]+ .

[0102] Synthesis of 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) using synthesis method A [ka]

[0103] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2): 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-neck flask, and the mixture was heated to 90 °C with stirring. Solid (NH)SO (25 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 2 h. Solid KSO (30 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 20 h. The reaction was monitored by LC-MS, showing remaining starting material. EtOAc (150 mL) and water (300 mL) were added to the mixture, stirred, and separated. The aqueous phase was again extracted with EtOAc (150 mL). The combined organic phase was washed with saturated aqueous sodium chloride (150 mL), concentrated, and the residue was purified by column chromatography (Et0Ac:Hexane = 0:1 to 1:5 to 1:2) to give the product (1.82 g, 10.5% yield). The remaining starting material was recovered (10.3 g, 68.7% yield). 1 H NMR (400 MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0104] 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): 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 one-neck flask. The mixture was purged with argon, stirred, and heated at reflux for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was concentrated, and the residue was purified by column chromatography to give the product (368 mg, 75% yield). 1 H NMR (400 MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J = 3.8 Hz, 1H), 7.51-7.44 (m, 2H), 6.99- 6.88 (m, 2H), 5.67 (d, J = 3.9 Hz, 1H), 3.80-3.63 (m, 2H), 2.84-2.42 (m, 10H), 2.39 (ddt, J = 11.4, 7.3, 3.7 Hz, 1H), 1.96 (dt, J = 12.2, 3.0 Hz, 2H), 1.70 (qd, J = 12.1, 4.0 Hz, 2H); MS (ESI): 556 [M+H] + .

[0105] 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): 5-Chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (166.76 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), (1H-indol-4-yl)boronic acid (53.12 mg, 0.33 mmol), dioxane / HO (10 mL / 2 mL), and Pd(dppf)Cl (22 mg) were added to a 50 mL single-neck flask. The mixture was purged with argon, rapidly heated to 105 °C, and incubated for 60 min. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (131 mg, 80% yield). MS (ESI): 545 [M+H] + .

[0106] 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)pyrazine-2-carboxamide (compound 511): 5-Chloro-6-(1H-indol-4-yl)-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (164 mg, 0.3 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (35 mg, 0.6 mmol), DMSO (5 mL), and 4 Å molecular sieves (200 mg, powder) were added to a 50 mL single-neck flask. The mixture was purged with argon and stirred at room temperature for 15 minutes. Next, 3-tetrahydro-2H-pyran-4-amine (33.4 mg, 0.33 mmol) was added, and the mixture was purged with argon, heated to 120 °C, and stirred for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (127 mg, 69.4% yield). 1H NMR (400 MHz, CDCl3) δ 10.81 (s, 1H), 8.39 (s, 1H), 7.62 (d, J = 8.9 Hz, 2H), 7.55-7.45 (m, 2H), 7.30 (td, J = 6.3, 5.7, 4.0 Hz, 3H), 6.92 (d, J = 8.9 Hz, 2H), 6.54 (d, J = 2.8 Hz, 1H), 5.25 (d, J = 7.2 Hz, 1H), 5.13 (s, 1H), 4.23-4.14 (m, 1H), 3.96 (d, J = 11.7 Hz, 2H), 3.69 (d, J = 11.9 Hz, 2H), 3.56-3.47 (m, 2H), 2.77-2.37 (m, 11H), 2.32 (s, 3H), 2.00 (dd, J= 26.0, 11.4 Hz, 4H), 1.72 (dd, J = 11.8, 3.8 Hz, 2H), 1.49-1.42 (m, 2H); MS (ESI): 610 [M+H] + .

[0107] Synthesis method B: Synthesis of Compound 19 (5-Methoxy-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide) using Synthesis Method B [ka] TIFF0007777544000045.tif61151

[0108] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2): 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-neck flask, and the mixture was heated to 90 °C with stirring. Solid (NH)SO (25 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 2 h. Solid KSO (30 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 20 h. The reaction was monitored by LC-MS, showing remaining starting material. EtOAc (150 mL) and water (300 mL) were added to the mixture, stirred, and separated. The aqueous phase was again extracted with EtOAc (150 mL). The combined organic phase was washed with saturated aqueous sodium chloride (150 mL), concentrated, and the residue was purified by column chromatography (Et0Ac:Hexane = 0:1 to 1:5 to 1:2) to give the product (1.82 g, 10.5% yield). The remaining starting material was recovered (10.3 g, 68.7% yield). 1 H NMR (400 MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0109] 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): 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 one-neck flask. The mixture was purged with argon, stirred, and heated at reflux for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was concentrated, and the residue was purified by column chromatography to give the product (368 mg, 75% yield). 1H NMR (400 MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J = 3.8 Hz, 1H), 7.51-7.44 (m, 2H), 6.99- 6.88 (m, 2H), 5.67 (d, J = 3.9 Hz, 1H), 3.80-3.63 (m, 2H), 2.84-2.42 (m, 10H), 2.39 (ddt, J = 11.4, 7.3, 3.7 Hz, 1H), 1.96 (dt, J = 12.2, 3.0 Hz, 2H), 1.70 (qd, J = 12.1, 4.0 Hz, 2H); MS (ESI): 556 [M+H] + .

[0110] 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): 5-Chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (167 mg, 0.30 mmol), anhydrous potassium phosphate (160 mg, 0.75 mmol), phenylboronic acid (40.23 mg, 0.33 mmol), dioxane / HO (10 mL / 2 mL), and Pd(dppf)Cl (22 mg) were added to a 50 mL single-neck flask. The mixture was purged with argon, rapidly heated to 105 °C, and incubated for 30 min. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (115 mg, 75.6% yield). 1H NMR (400 MHz, CDCl3) δ: 10.74 (s, 1H), 7.70 (d, J = 6.5 Hz, 3H), 7.56 (d, J = 8.5 Hz, 2H), 7.44 (p, J = 6.8 Hz, 3H), 6.94 (d, J = 8.6 Hz, 2H), 5.67 (s, 1H), 3.70 (d, J = 11.9 Hz, 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 = 12.4 Hz, 2H), 1.67 (tt, J= 12.5, 6.8 Hz, 2H); LC-MS: 506 [M+H] + .

[0111] 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): 5-Chloro-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenylpyrazine-2-carboxamide (50 mg, 0.10 mmol), DMF (5 mL), and 4 Å molecular sieves (200 mg, powder) were added to a 50 mL single-neck flask. The mixture was purged with argon and stirred at room temperature for 15 minutes. Sodium methoxide (16 mg, 0.3 mmol) was then added, and the mixture was purged with argon, heated to 80° C., and stirred for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (21 mg, 42% yield). 1H NMR (400 MHz, CDCl3) δ: 10.81 (s, 1H), 7.99-7.87 (m, 2H), 7.65 (d, J = 4.4 Hz, 1H), 7.61-7.55 (m, 2H), 7.41 (d, J = 7.8 Hz, 2H), 6.94 (dd, J = 9.0, 3.6 Hz, 2H), 5.42-5.33 (m, 1H), 4.05 (s, 3H), 3.70 (d, J = 12.0 Hz, 2H), 2.78-2.41 (m, 11H), 2.41-2.32 (m, 1H), 2.30 (s, 3H), 1.94 (d, J = 12.4 Hz, 2H), 1.69 (qd, J = 11.8, 3.7 Hz, 2H); MS (ESI): 502 [M+H] + .

[0112] Synthesis method C: Synthesis of Compound 116 (3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-6-phenyl-5-((tetrahydro-2H-pyran-4-yl)oxo)pyrazine-2-formamide) using Synthetic Method C [ka]

[0113] Step 1: Synthesis of compound 3,5-dichloro-6-iodopyrazine-2-carboxamide (compound int_2) 3,5-Dichloro-2-iodopyrazine (15 g, 54.57 mmol) and formamide (300 mL) were added to a 500 mL single-neck flask, and the mixture was heated to 90 °C with stirring. Solid (NH)SO (25 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 2 h. Solid KSO (30 g, 109.1 mmol) was added in batches, and the mixture was stirred at 90 °C for 20 h. The reaction was monitored by LC-MS, showing remaining starting material. EtOAc (150 mL) and water (300 mL) were added to the mixture, stirred, and separated. The aqueous phase was again extracted with EtOAc (150 mL). The combined organic phase was washed with saturated aqueous sodium chloride (150 mL), concentrated, and the residue was purified by column chromatography (Et0Ac:Hexane = 0:1 to 1:5 to 1:2) to give the product (1.82 g, 10.5% yield). The remaining starting material was recovered (10.3 g, 68.7% yield). 1 H NMR (400 MHz, CDCl3) δ: 7.28 (s, 1H), 5.78 (s, 1H); MS (ESI): 317 [M+H] + .

[0114] 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): 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 one-neck flask. The mixture was purged with argon, stirred, and heated at reflux for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was concentrated, and the residue was purified by column chromatography to give the product (368 mg, 75% yield). 1H NMR (400 MHz, CDCl3) δ: 10.69 (s, 1H), 7.53 (d, J = 3.8 Hz, 1H), 7.51-7.44 (m, 2H), 6.99- 6.88 (m, 2H), 5.67 (d, J = 3.9 Hz, 1H), 3.80-3.63 (m, 2H), 2.84-2.42 (m, 10H), 2.39 (ddt, J = 11.4, 7.3, 3.7 Hz, 1H), 1.96 (dt, J = 12.2, 3.0 Hz, 2H), 1.70 (qd, J = 12.1, 4.0 Hz, 2H); MS (ESI): 556 [M+H] + .

[0115] 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)pyrazine-2-carboxamide (compound int_5): 5-Chloro-6-iodo-3-((4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrazine-2-carboxamide (150 mg, 0.27 mmol), anhydrous potassium carbonate (186 mg, 1.35 mmol), anhydrous potassium fluoride (31 mg, 0.54 mmol), DMSO (5 mL), and 4 Å molecular sieves (200 mg, powder) were added to a 50 mL single-neck flask. The mixture was purged with argon and stirred at room temperature for 15 minutes. Next, 3-tetrahydro-2H-pyran-4-amine (32 mg, 0.32 mmol) was added, and the mixture was purged with argon, heated to 120° C., and stirred for 2 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (110 mg, 65.7% yield). 1H NMR (400 MHz, 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, 3.7 Hz, 3H), 3.67 (d, J = 12.0 Hz, 2H), 3.51 (td, J = 11.6, 2.2 Hz, 2H), 2.84-2.50 (m, 10H), 2.44 (d, J= 11.4 Hz, 1H), 2.37 (s, 3H), 2.11-1.89 (m, 4H), 1.78-1.51 (m, 4H); MS (ESI): 621 [M+H] + .

[0116] 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)pyrazine-2-carboxamide (compound 116): 6-Iodo-3-((4-(4-4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-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 hours. After completion of the reaction as indicated by LC-MS, the mixture was cooled and purified by column chromatography to give the product (32 mg, 57.4% yield). 1H NMR (400 MHz, CDCl3) δ: 10.57 (s, 1H), 7.55-7.48 (m, 2H), 6.90-6.82 (m, 2H), 5.51 (d, J = 7.3 Hz, 1H), 5.06 (s, 1H), 4.13-4.04 (m, 1H), 4.02-3.96 (m, 2H), 3.64 (d, J= 11.7 Hz, 2H), 3.55 (td, J = 11.5, 2.3 Hz, 2H), 2.87 (t, J = 5.3 Hz, 4H), 2.71-2.58 (m, 5H), 2.48 (s, 3H), 2.36 (t, J = 11.4 MS (ESI): 578 [M+H] + .

[0117] Example 1: Synthesis of Compound 1-645 Target compounds 1 to 18, compounds 20 to 115, compounds 117 to 135, and compounds 140 to 645 in Table 2 were obtained using synthesis method A, synthesis method B, or synthesis method C, while changing the starting material.

[0118] The LC-MS analysis process is as follows. Equipment: Agilent 6125B Chromatography column: Core-shell 2.7 μm 4.3 × 50 mm Column temperature: 30℃ Wavelength: 254nm / 214nm Mobile phase A: HO (0.1% formic acid) Mobile phase B: Acetonitrile (0.1% formic acid) gradient

[0119] [Table 1]

[0120] [Table 2-1] Table 2-2 Table 2-3 Table 2-4 Table 2-5 Table 2-6 Table 2-7 Table 2-8 Table 2-9 Table 2-10 Table 2-11 Table 2-12 Table 2-13 Table 2-14 Table 2-15 Table 2-16 Table 2-17 Table 2-18 Table 2-19 Table 2-20 Table 2-21 Table 2-22 Table 2-23 Table 2-24 Table 2-25 Table 2-26 Table 2-27 Table 2-28 Table 2-29 Table 2-30 Table 2-31 Table 2-32 Table 2-33 Table 2-34 Table 2-35 Table 2-36 Table 2-37 Table 2-38 Table 2-39 Table 2-40 Table 2-41 Table 2-42 Table 2-43 Table 2-44 Table 2-45 Table 2-46 Table 2-47 Table 2-48 Table 2-49 Table 2-50 Table 2-51 Table 2-52 Table 2-53 Table 2-54 Table 2-55 Table 2-56 Table 2-57 Table 2-58 Table 2-59 Table 2-60 Table 2-61 Table 2-62 Table 2-63 Table 2-64 Table 2-65 Table 2-66 Table 2-67 Table 2-68 Table 2-69 Table 2-70 Table 2-71 Table 2-72 Table 2-73 Table 2-74 Table 2-75 Table 2-76 Table 2-77 Table 2-78

[0121] Table 3-1 Table 3-2 Table 3-3 Table 3-4 Table 3-5 Table 3-6 Table 3-7 [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11] [Table 3-12] [Table 3-13]

[0122] Example 2: Inhibitory activity assay of compounds disclosed herein against EGFR(del19 / T790M / C797S), EGFR(L858R / T790M / C797S) or EGFR(WT) enzymes The inhibitory effect of compounds on the enzyme activity of EGFR(del19 / T790M / C797S), EGFR(L858R / T790M / C797S), or EGFR(WT) was determined using HTRF. The procedure was as follows.

[0123] WT or mutant EGFR protein was incubated with serially diluted compounds at 28°C for 10 minutes, followed by the addition of biotin-labeled general tyrosine kinase (TK) substrate and ATP. The mixture was incubated at room temperature for 40 minutes to allow for reaction. After the reaction was complete, Eu3+-Cryptate-labeled antibody against TK and streptavidin-XL665 were added, and the mixture was incubated at room temperature for 60 minutes. Emissions at 615 nm and 665 nm were detected, and the 665 / 615 ratio was calculated to quantify the level of TK substrate phosphorylation. The compound inhibition rate and IC 50 was calculated in comparison with the control group. The results are shown in Table 4 below. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] [Table 4-6] [Table 4-7]

[0124] As can be seen from the data in Table 4, the compounds disclosed herein have superior inhibitory activity against EGFR(del19 / T790M / C797S) and EGFR(L858R / T790M / C797S) enzymatic activity and superior selectivity against EGFR(WT).

[0125] Example 3: Antiproliferative activity of compounds disclosed herein against Ba / F3 (EGFRdel19 / T790M / C797S) triple mutant cells and A431 (EGFRWT) cells Three thousand Ba / F3 cells or 2000 A431 cells bearing EGFR (del19 / T790M / C797S) were seeded in a 384-well plate. One day later, serially diluted compounds were added (up to 500 nM for Ba / F3 cells and up to 10 μM for A431 cells). Three days after compound addition, cell proliferation was assessed by adding Cell TiterGlow, and the cell growth inhibition rate and IC40 induced by the compounds were determined. 50 The values ​​were calculated and the results are shown in Table 5 below.

[0126] [Table 5-1] [Table 5-2] [Table 5-3]

[0127] As can be seen from the data in Table 5, Ba / F3(EGFR del19 / T790M / C797S The antiproliferative activity of most compounds disclosed herein against Ba / F3 (EGFR) triple mutant cells was less than 100 nM, but del19 / T790M / C797S The antiproliferative activity of gilteritinib against Ba / F3 (EGFR triple mutant) cells is greater than 500 nM, and when Y is aryl, heteroaryl, or heterocycloalkyl, the compound del19 / T790M / C797S ) are shown to have strong antiproliferative activity against triple mutant cells.

[0128] Example 4: In Vivo Pharmacodynamics Study - Mouse H1975 Subcutaneous Xenograft Tumor Model 5 × 10 EGFR T790M mutation-carrying mice were placed on the left dorsum of BALB / c nude mice. 6 The cells were subcutaneously transplanted. The tumor size was 100-150 mm. 3 After the mice reached adulthood, they were randomly divided into the following groups and administered compound 511 intragastrically once daily: Group 1: vehicle control, Group 2: compound 511 (60 mg / kg), and Group 3: compound 511 (80 mg / kg). Tumor volumes were measured twice a week and at the end of treatment. The tumor growth inhibition of the compound was calculated according to the following formula: tumor growth inhibition (TGI) = 1 - (tumor volume in the treatment group on day 28 - tumor volume in the treatment group on day 1) / (tumor volume in the vehicle control group on day 28 - tumor volume in the treatment group on day 1). The results are shown in Figure 1 and Table 6.

[0129] [Table 6]

[0130] As can be seen from Figure 1 and Table 6, compound 511 was able to inhibit tumor growth at doses of 60 mg / kg and 80 mg / kg in the H1975 mouse subcutaneous xenograft tumor model harboring the EGFR T790M mutation.

[0131] Example 5: In Vivo Pharmacodynamic Study - Mouse PC9 (EGFR Del19 / T790M / C797S) Subcutaneous Xenograft Tumor Model 5 × 10 EGFR Del19 / T790M / C797S-overexpressing cells were injected into the left dorsal region of BALB / c nude mice. 6 PC9 cells were subcutaneously transplanted. The tumor size was 100-150 mm. 3 After the mice reached adulthood, they were randomly divided into the following groups and administered compound 511 intragastrically once daily: Group 1: vehicle control, Group 2: compound 511 (60 mg / kg), and Group 3: compound 511 (80 mg / kg). Tumor volumes were measured twice a week and at the end of treatment. The tumor growth inhibition of the compound was calculated according to the following formula: tumor growth inhibition (TGI) = 1 - (tumor volume in the treatment group on day 28 - tumor volume in the treatment group on day 1) / (tumor volume in the vehicle control group on day 28 - tumor volume in the treatment group on day 1). The results are shown in Figure 2.

[0132] [Table 7]

[0133] As can be seen from Figure 2 and Table 7, compound 511 was able to inhibit tumor growth at doses of 60 mg / kg and 80 mg / kg in a PC9 mouse subcutaneous xenograft tumor model overexpressing EGFR Del19 / T790M / C797S.

Claims

1. A compound having a structure of general formula (1), or an optical isomer, pharmaceutically acceptable salt, hydrate, or solvate thereof: 【Chemistry 1】 (wherein, in general formula (1), Y is, 【Chemistry 2】 【change】 and L １ is —NH—, X is 【Transformation 3】 and R １ is -N(CH ３ ) ２ , -CH ２ - a 6- to 11-membered heterocycloalkyl or a 6- to 11-membered heterocycloalkyl, wherein said heterocycloalkyl is 【Chemistry 4】 and -H, -CH 3 , 【Transformation 5】 -N(CH3)2, 【Transformation 6】 and -CD3, which may be optionally substituted with one or more groups; L ２ is —O—, —NH— or a chemical bond, R ２ represents C1-C6 alkyl, C3-C14 cycloalkyl, C6-C14 aryl, 3- to 4-membered heterocycloalkyl, 【Transformation 7】 or a 6- to 11-membered heterocycloalkyl, wherein said alkyl, said cycloalkyl, said aryl, said heterocycloalkyl, 【Transformation 8】 is -H, halogen, -R ４ , -(CH ２ ) ｎ OR4, -(CH ２ ) ｎ NR ４ R5, -OR ４ , -NR ４ R ５ , -CN, -C(O)NR ４ R ５ , -NR ５ C(O)R ４ , -NR ５ S (O) ２ R ４ , -S(O) ｐ R ４ and -S(O) ２ NR ４ R ５ and R ４ and R ５ are each independently —H, C1-C6 alkyl, or C3-C14 cycloalkyl; R ０ is C1-C6 alkyl or C3-C14 cycloalkyl, 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.

2. In general formula (1), R １ is -H, -N(CH ３ ) ２、 【Chemistry 9】 2. The compound of claim 1, wherein:

3. In general formula (1), L ２ When is -NH-, R ２ teeth 【Chemistry 10】 2. The compound of claim 1, wherein:

4. In general formula (1), L ２ When is -O-, R ２ teeth, 【Chemistry 11】 2. The compound of claim 1, wherein:

5. In general formula (1), L ２ When is a chemical bond, R ２ teeth, 【Chemistry 12】 2. The compound of claim 1, wherein:

6. The compound has the following structure: 【Chemistry 13】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 or an optical isomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, of the compound of claim 1 having one of the following formulas:

7. A pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier and, as an active ingredient, the compound of claim 1 or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof.

8. 10. Use of the compound of claim 1, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, in preparing a medicament for treating a disease associated with an EGFR mutation.

9. 8. The pharmaceutical composition according to claim 7 for treating, regulating and / or preventing a disease associated with an EGFR mutein, comprising:

10. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof.

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