Compounds for use as kinase inhibitors and their uses - Patents.com

Abstract

The present invention belongs to the field of drug synthesis of heterocyclic compounds with nitrogen atom as heterocyclic atom, specifically relates to a compound used as a kinase inhibitor and its use, and also relates to the preparation of the free base crystal form of the compound. The compound used as the kinase inhibitor is a compound represented by formula 1, or its deuterated derivative, or a pharma- ceutically acceptable salt, solvate or prodrug. The above compounds have been demonstrated by biological activity experiments to have good inhibitory activity against exon 20 insertion mutation of EGFR and Her2, exon 19 deletion of EGFR, and exon 21 point mutation, and are useful as raw materials for related drugs.

Description

[Technical field]

[0001] The present invention is in the field of drug synthesis of heterocyclic compounds having a nitrogen atom as a heterocyclic atom, and specifically relates to compounds used as kinase inhibitors and their use. [Background technology]

[0002] EGFR belongs to the receptor tyrosine kinase (RTK) family, including EGFR / ERBB1, HER2 / ERBB2 / NEU, HER3 / ERBB3, and HER4 / ERBB4. EGFR activates its tyrosine kinase activity by homodimerization or heterodimerization, and phosphorylates its substrates, activating several downstream pathways associated with it in cells, such as the PI3K-AKT-mTOR pathway involved in cell survival and the RAS-RAF-MEK-ERK pathway involved in cell proliferation. EGFR mutations and amplifications cause activation of heterodimer kinases, which leads to the development of many human diseases, such as malignant tumors. For example, among non-small cell lung cancer patients, more than 10% of patients in the United States have EGFR mutations, and the proportion of patients with EGFR mutations in Asian patients reaches nearly 50%. At the same time, the incidence rate of HER2 mutations in non-small cell lung cancer patients is about 2-4%.

[0003] EGFR mutations mainly include deletions, insertions and point mutations, among which exon 19 deletions and exon 21 L858R point mutations account for nearly 90% of EGFR mutations. For patients with tumors with such EGFR mutations, the currently available EGFR-TKIs include the first generation Iressa, Tarceva, and Commana, the second generation Afatinib and Dacomitinib, and the third generation Osimertinib. The other 10% of EGFR mutations mainly involve EGFR exons 18 and 20, and EGFR exon 20 insertion mutations account for about 9% of all EGFR mutations. For patients with tumors with HER2 mutations, the most common HER2 mutation is the HER2 exon 20 insertion mutation.

[0004] TAK-788 has a therapeutic effect on EGFR and HER2 exon 20 insertion mutations, and the compound has already been marketed in the United States, and the reported clinical trial results show an objective remission rate of 43%. Recently, it has been reported that DZD9008 is effective in treating late-stage non-small cell lung cancer with EGFR or HER2 mutations, but the objective remission rate of EGFR exon 20ins is 40%, and the therapeutic effect is unsatisfactory. Summary of the Invention [Problem to be solved by the invention]

[0005] In order to meet the needs of EGFR and HER2 mutation patients in clinical practice, especially EGFR, HER2 exon 20 insertion mutation patients, WO2021180238 discloses a series of excellent compounds with activity against EGFR and HER2 exon 20 insertion mutations, as well as EGFR exon 19 deletion and exon 21 L858R point mutation, which have high potential potential for development as drugs to treat related diseases. Among them, for compounds with tripentamine structure, further research shows that the configuration of the tripentamine structure is very important, and when it goes from trans to cis, the drug effect in the body is better, and it also has good therapeutic effect on different mutations of related diseases. In the subsequent research and development, in order to find a free base compound with more stability and higher bioavailability, the present invention has studied and screened polymorphisms of free base compounds.

[0006] The object of the present invention is to provide a compound which is used as a kinase inhibitor, which belongs to cis-tripentamine structure and has good inhibitory activity against EGFR, HER2 exon 20 insertion mutation, EGFR exon 19 deletion and exon 21 point mutation. The compound of the present invention has better drug effect and wider applicability than the corresponding trans-structure compound in patent WO2021180238.

[0007] A second object of the present invention is to provide the use of said compounds in the manufacture of a medicament for the treatment of diseases associated with EGFR mutations and / or HER2 mutations. The present invention also relates to the preparation of crystalline forms of the free base of the compounds, and polymorphic studies are carried out to identify practical crystalline forms with high stability and higher bioavailability. [Means for solving the problem]

[0008] To achieve the above objectives, the technical solutions used in the present invention are as follows: A compound for use as a kinase inhibitor, the compound being represented by formula 1, or a deuterated form, or a pharma- ceutically acceptable salt, solvate or prodrug thereof. [ka] (In formula 1, X is selected from CH and N. R1 is [ka] and R5 is H, a C1-C3 alkyl group, or a C1-C3 fluoroalkyl group. R 20 , R 21 , R 22 are each independently selected from a methyl group or a deuterated methyl group. R3 is selected from a C1-C3 alkyl group and a C1-C3 haloalkyl group. R 40 , R 41 , R 42 are each independently selected from H, D, and F.

[0009] Biological activity experiments have demonstrated that the above compounds have good inhibitory activity against EGFR and HER2 mutations, and are useful as raw materials for related drugs. Based on the above drug substance, a "pharmaceutical acceptable salt" of the drug substance is a salt that is pharmaceutical acceptable and possesses the desired pharmacological activity of the parent compound. Such salts include the following:

[0010] Acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, etc., and typical inorganic acid salts are selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfate salts, hydrogen sulfate salts, nitrate salts, phosphate salts, and acid phosphate salts. Acid addition salts formed with organic acids, such as formic acid, acetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluene ... These include salts of toluenesulfonic acid, camphorsulfonic acid, gluceptic acid, 4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, etc., or salts in which the acidic protons present in the parent compound are coordinated with an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, trometamol, N-methylglucamine, etc.). Exemplary organic acid salts are selected from the group consisting of formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, hydroxyethanesulfonate, benzenesulfonate, salicylate, picrate, glutamate, ascorbate, camphorate, and camphorsulfonate. It is readily understood that the pharma- ceutically acceptable salts are non-toxic.

[0011] A solvate is a compound that contains a solvent, for example, a hydrate, a dimethylsulfoxide solvate, and the like. Prodrugs are compounds which, upon chemical conversion by metabolic or chemical processes, produce the compounds, salts, or solvates of the compounds of the present invention when used to treat the relevant disease.

[0012] Preferably, the compound used as the kinase inhibitor is a compound represented by formula 2, or a deuterated form, or a pharma- ceutically acceptable salt, solvate or prodrug thereof. [ka] More preferably, in formula 2, X is selected from CH and N, R is selected from -CH, -CHCH, -CHCF, and R 40 , R 41 are both H and R 42 is selected from H or F.

[0013] Preferably, the compound used as the kinase inhibitor is a compound represented by formula 3, or a deuterated form thereof, or a pharma- ceutically acceptable salt, solvate or prodrug thereof. [ka] In formula 3, X is selected from CH, R3 is -CH3, -CH2CH3, -CH2CF3, R 40 , R 41 are both H and R 42 is selected from H or F, and R5 is selected from -CH3, -CF3.

[0014] Preferably, the compound of formula 1 is: [ka]

[0015] Use of a compound used as a kinase inhibitor as described above for the manufacture of a drug for treating diseases associated with EGFR mutations and / or HER2 mutations. The above compounds have good inhibitory activity against EGFR and HER2 exon 20 insertion mutations, and also have good inhibitory activity against EGFR exon 19 deletions and exon 21 point mutations.

[0016] Preferably, the EGFR mutation and / or HER2 mutation includes one or more of the following combinations: EGFR exon 20 insertion mutation, HER2 exon 20 insertion mutation, EGFR exon 19 deletion, EGFR exon 20 point mutation, and EGFR exon 21 point mutation. Biological activity experiments have demonstrated that the compound has good inhibitory effects on the above types of mutation.

[0017] More preferably, the EGFR mutation and / or HER2 mutation is selected from EGFR Del 19 / T790M / C797S mutation, EGFR L858R / T790M / C797S mutation, and the compound has good inhibitory effect on the above mutation types.

[0018] Preferably, the disease is cancer caused by the EGFR mutation and / or HER2 mutation.The above compound may be used in combination with other drugs to treat cancer.The other drugs used in combination may be ERK inhibitors or MEK inhibitors. Preferably, the compound is in a crystalline form, an amorphous form or a solvate, and the solvent contained in the solvate is a non-aqueous solvent or a mixed solvent consisting of a non-aqueous solvent and water.

[0019] In order to further improve the stability and activity of the compound, a study on the crystal form of a selected compound represented by general formula 1 was carried out. Furthermore, a screening study of crystal polymorphism was carried out on the compound of Example 1 in the present invention to find a stable and reliable crystal form, so that the drug can exert a better effect in clinical treatment while ensuring the stability of the quality of the compound. The product of Example 1 was found to be a crystal with good crystallinity, and is an anhydrous crystal form, named free base crystal form I. The compound of Example 1 has the compound represented by formula A, [ka] The crystalline form is free base crystalline form I, and the powder X-ray diffraction spectrum of said crystalline form has characteristic diffraction peaks at 9.76°±0.2°, 10.45°±0.2°, 16.54°±0.2°, 18.66°±0.2°, 20.07°±0.2°, and 25.90°±0.2°.

[0020] Additionally, the powder X-ray diffraction spectrum of the free base crystalline Form I contains characteristic diffraction peaks at 9.07°±0.2°, 9.76°±0.2°, 10.45°±0.2°, 11.53°±0.2°, 11.80°±0.2°, 12.91°±0.2°, 13.79°±0.2°, 14.67°±0.2°, 15.08°±0.2°, 15.63°±0.2°, 16.54°±0.2°, 17.50°±0.2°, 18.66°±0.2°, 20.07°±0.2°, 21.10°±0.2°, 23.29°±0.2°, 24.16°±0.2°, and 25.90°±0.2°.

[0021] The free base crystalline Form I of the compound of Example 1 can be prepared by methods such as suspension crystallization, crude solvent precipitation, high-temperature circulation and evaporative crystallization in many solvents, including, but not limited to, dichloromethane, 1,4-dioxane, dichloroethane, methyl-t-butyl ether, N-methylpyrrolidone, ethyl acetate, acetone, methanol, dimethylsulfoxide, isopropyl acetate, butanone, cyclohexane, tetrahydrofuran, water, acetonitrile, isopropanol, ethanol, n-heptane, or a mixture of two or more thereof.

[0022] In the present invention, the compound of Example 1 was found to have two anhydrous crystalline forms (free base crystalline Form I and free base crystalline Form V) and three solvates, free base crystalline Form II is a solvate with ethanol, free base crystalline Form III is a solvate with isopropanol, and free base crystalline Form IV is a solvate with dichloroethane and water, and all of these solvates were characterized. Competitive suspension experiments were carried out against the two anhydrous crystalline forms, free base crystalline Form I and free base crystalline Form V, and it was predicted that free base crystalline Form I is a thermodynamically stable crystalline form and is more suitable for further development.

[0023] The evaluation of the crystal form of the free base crystalline form I was carried out, including dry grinding, wet grinding, tableting (30 MPa), stability and hygroscopicity. After 5 min of dry grinding, it became amorphous. After 5 min of wet grinding with water, it remained almost unchanged, and after 5 min of wet grinding with ethanol, the crystallinity was slightly decreased. In the tableting test, the crystallinity was decreased to a certain extent when the pressure was 30 MPa. In terms of stability, the free base crystalline form I was relatively stable at different temperatures and humidities, and the liquid phase detection results did not change. The dynamic vapor sorption (DVS) test showed that at 80% RH, the weight increase due to water absorption was 0.66%, and the free base crystalline form I was slightly hygroscopic. In other words, the free base crystalline form I is a relatively stable anhydrous crystalline form, has stable solid properties, and is slightly hygroscopic, which is useful for subsequent drug development. [Brief description of the drawings]

[0024] [Figure 1] FIG. 1 shows the change in tumor volume (mm3) in the LU0387 model. [Diagram 2] FIG. 2 shows the change in tumor volume (mm3) in the Ba / F3 EGFR D770_N771 ins SVD model. [Diagram 3] FIG. 3 is a single crystal diffraction spectrum of the single crystal of Example 1. [Figure 4] FIG. 4 is an XRPD spectrum of free base crystalline Form I. [Diagram 5]FIG. 5 is an overlay of DSC and TGA of free base crystalline Form I. [Figure 6] FIG. 6 shows the XRPD spectrum of an amorphous sample obtained by the dry grinding method. [Figure 7] FIG. 7 is an XRPD spectrum of the free base crystalline form II. [Figure 8] FIG. 8 is an overlay of DSC and TGA of the free base crystalline Form II. [Figure 9] FIG. 9 is an XRPD spectrum of the free base crystalline Form III. [Figure 10] FIG. 10 is an overlay of DSC and TGA of the free base crystalline Form III. [Figure 11] FIG. 11 is an XRPD spectrum of the free base crystalline Form IV. [Figure 12] FIG. 12 is an overlay of DSC and TGA of the free base crystalline Form IV. [Figure 13] FIG. 13 is an XRPD spectrum of free base crystalline form V. [Figure 14] FIG. 14 is an overlay of DSC and TGA of free base crystalline form V. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] Compared with the trans-tripentamine structure in Patent WO2021180238, the present invention demonstrates that the cis-tripentamine structure has better medicinal effect in the body and also has excellent therapeutic effect in different mutations of related diseases.

[0026] In the preparation method of the compound of the present invention when X is CH, the reaction pathway is as follows. [ka]

[0027] The process includes the following steps: (1) Compound A and compound B are subjected to a Friedel-Crafts reaction in an organic solvent to obtain compound C; (2) Compound C and compound D are subjected to a substitution reaction in an organic solvent under an acid catalyst to obtain compound E; (3) Compound E and compound F are subjected to a substitution reaction in an organic solvent under the catalysis of a base to obtain compound G; (4) reducing compound G to obtain compound H; (5) Compound H is subjected to a condensation reaction in the presence of a base catalyst to obtain a product.

[0028] R1 is [ka] and R5 is -CF3, the synthesis of compounds of formula 1 may be via the following reaction pathway: [ka]

[0029] The process includes the following steps: (1) Compound a and compound b are subjected to a substitution reaction in an organic solvent to obtain compound c; (2) Hydrolyzing compound c to obtain compound d; (3) Compound d is subjected to a condensation reaction to obtain compound e; (4) Compound e is deprotected to obtain compound f; (5) Compound f is subjected to a ring-closing reaction with trifluoroacetic anhydride in the presence of a base catalyst to obtain compound g; (6) reducing compound g to obtain compound h; (7) Compound h is subjected to a condensation reaction in the presence of a base catalyst to obtain a product.

[0030] In the preparation method of the compound of the present invention when X is N, the reaction pathway is as follows. [ka]

[0031] The method includes the following steps: Compound A' and Compound B' are subjected to a substitution reaction to obtain a product. Here, compound A' is obtained by synthesis with reference to WO2021180238, and compound B' is obtained by the Friedel-Crafts reaction.

[0032] Hereinafter, the implementation process of the present invention will be described in detail with specific examples. 1. Specific Examples of Synthesis of Compounds Used as Kinase Inhibitors

[0033] Example 1 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0034] The synthetic route of the compound of this example is as follows. [ka]

[0035] Synthesis of compound 3: In a 250mL three-neck flask, under the protection of nitrogen gas, compound 1 (2.01g, 8.5mmol) was added, dissolved completely in 100mL of tetrahydrofuran, anhydrous aluminum trichloride (2.26g, 17mmol) was added, and the mixture was stirred at 70°C for 1 hour. Compound 2 (1.34g, 10.2mmol) was added dropwise, and after the addition was completed, the mixture was reacted at 70°C, and the reaction was monitored by sampling on a plate. After 4 hours, the reaction was almost complete. After processing, 1.31g of product was obtained, and the yield was 46.4%.

[0036] Synthesis of compound 5: Compound 3 (1.30 g, 3.9 mmol), compound 4 (0.88 g, 4.68 mmol), and p-toluenesulfonic acid (1.35 g, 7.8 mmol) were added to 65 mL of 1,4-dioxane, and the mixture was heated to 80° C. under N2 protection and reacted overnight. The reaction was monitored by sampling on a plate, and the reaction was completed and treated to obtain 1.50 g of product, with a yield of 79.7%.

[0037] Synthesis of compound 7: Compound 5 (6.0 g, 12.5 mmol), compound 6 (6.3 g, 100 mmol) and N,N-diisopropylethylamine (3.2 g, 50 mmol) were added to 120 mL of N,N-dimethylacetamide, reacted at 90° C. overnight, sampled on a plate for monitoring, and treated after the reaction was complete to obtain 8.0 g of the product. 1 H NMR (400 MHz, chloroform-d) δ 9.53 (s, 1H), 8.97 - 8.75 (m, 1H), 7.96 (s, 1H), 7.84 (s, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.38 (d, J = 8.2 Hz, 1H), 7.29 (d, J = 7.2 Hz, 1H), 7.20 (t, J = 7.6 Hz, 1H), 6.81 (s, 1H), 5.09 (p, J = 6.4 Hz, 1H), 4.02 (s, 3H), 3.92 (s, 3H), 3.84 - 3.63 (m, 1H), 3.02 (t, J = 6.4 Hz, 1H), 2.84 (s, 3H), 2.70 (s, 3H), 2.43 (s, 2H), 2.32 (s, 1H), 1.27 (d, J = 7.2 Hz, 2H), 1.15 (d, J = 6.4 Hz, 6H).

[0038] Synthesis of compound 8: Compound 7 (7.7 g, 41.6 mmol) and palladium carbon (2.4 g, wet palladium carbon 55%) were placed in a mixed solvent of 100 mL of methanol and 100 mL of ethyl acetate, and reacted at room temperature for 4 hours in a hydrogen gas atmosphere. The mixture was sampled and monitored on a plate. After the raw materials were completely reacted, the mixture was treated to obtain 7.0 g of the product.

[0039] Synthesis of Example 1: Compound 8 (7.00g, 12.6mmol) and triethylamine (3.82g, 37.8mmol) were dissolved in dichloromethane, and the temperature was lowered to 0°C under the protection of nitrogen gas. Then, a solution of acrylic chloride (1.72g, 18.9mmol) in dichloromethane was added dropwise, and the mixture was reacted for 1 hour while maintaining the temperature at 0°C. The mixture was sampled and monitored on a plate, and the raw material was completely reacted before being processed. That is, an aqueous solution of sodium bicarbonate and dichloromethane were added, and the mixture was separated by stirring. The aqueous phase was further extracted once with dichloromethane, and the organic phase was combined, dried, spun dry, and passed through a column. The obtained crude product was first dissolved in a small amount of dichloromethane, and then petroleum ether was added dropwise to precipitate a large amount of solid. The mixture was filtered, and the cake was directly put into methanol, mixed, and dried, and finally 3.5g of the product was obtained. [M+H] + :610.8; 1 H NMR (400 MHz, chloroform-d) δ 10.38 (s, 1H), 9.60 (s, 1H), 8.87 (s, 1H), 8.61 (s, 1H), 7.89 (s, 1H), 7.58 (s, 1H), 7.32 (d, J = 8.2 Hz, 1H), 7.20 (t, J = 7.6 Hz, 1H), 7.13 (t, J = 7.6 Hz, 1H), 6.80 (s, 1H), 6.44 (m, 2H), 6.32(m,1H), 5.68 (m, 1H), 5.00 (p, J = 6.2 Hz, 1H), 3.94 (s, 3H), 3.88 (s, 3H), 3.06 (s, 2H), 2.66 (s, 3H), 2.61 (m, 3H), 2.36 (s, 3H), 1.67 (s, 2H), 1.03 (d, J = 6.4 Hz, 6H).

[0040] With reference to Example 1, three deuterated compounds were synthesized in Examples 2, 3, and 4, which are specifically shown in Table 1 below. Table 1. Structures and Characterization of Compounds of Examples 2-4 [Table 1]

[0041] Example 5 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0042] The synthetic route of the compound of this example is as follows. [ka]

[0043] The synthesis process is as follows. Synthesis of compound 2: In a 2000mL single-neck flask, compound 1 (54.6g, 303mmol), acetohydrazide (26.64g, 395mmol), and 1000mL of 1N aqueous sodium hydroxide solution were added, and the mixture was reacted at 80℃ for 4 hours, resulting in the precipitation of a large amount of solid. The temperature was lowered, and 80mL of concentrated hydrochloric acid was added, followed by stirring at a low temperature of about 0℃ for 30 minutes, followed by suction filtration, stirring the cake once with water, and vacuum drying at 55℃ overnight to obtain 32g of product. 1 H NMR (400 MHz, DMSO-d6) δ 11.75 (s, 1H), 11.59 (s, 1H), 8.13 (s, 1H), 2.52 (s, 3H).

[0044] Synthesis of compound 3: In a 2000mL single-neck flask, compound 2 (50.0g, 258mmol) was placed, followed by 1500mL of toluene, phosphoryl chloride (237g, 1550mmol), and N,N-diisopropylethylamine (133g, 1031mmol) at room temperature, generating a white mist. The mixture was heated to 80°C and stirred overnight under nitrogen gas protection. The next day, the mixture was cooled and treated to obtain 30g of the product. 1 H NMR (400 MHz, chloroform-d) δ 9.19 (s, 1H), 2.71 (s, 3H).

[0045] Synthesis of compound 5: In a 2000mL single-neck flask, compound 3 (30.0g, 130mmol) was placed and dissolved completely in 1500mL of 1,2-dichloroethane. Anhydrous aluminum trichloride (29.3g, 220mmol) was added and stirred at room temperature for 30 minutes. The temperature was lowered to 0°C, compound 4 (22.1g, 169mmol) was added dropwise, and the mixture was stirred at low temperature for 30 minutes, then heated to 60°C for reaction, and sampled on a plate for monitoring. After reacting for 4 hours, the mixture was cooled and processed to obtain 20.0g of product. 1 H NMR (400 MHz, chloroform-d) δ 8.84 (s, 1H), 8.02 (dt, J = 7.8, 1.0 Hz, 1H), 7.97 (s, 1H), 7.40-7.27 (m, 3H), 3.86 (s, 3H), 2.50 (s, 3H).

[0046] Synthesis of compound 7: Compound 5 (2 g, 6.15 mmol), compound 6 (1.37 g, 7.38 mmol), and p-toluenesulfonic acid (2.11 g, 12.3 mmol) were added to 100 mL of dioxane, and the mixture was heated to 80° C. under nitrogen gas protection and reacted overnight. The mixture was sampled on a plate for monitoring, and after the reaction was completed, it was treated to obtain 0.65 g of the product.

[0047] Synthesis of compound 9: Compound 7 (950 mg, 2 mmol), compound 8 (1.0 g, 8 mmol) and N,N-diisopropylethylamine (516 mg, 4 mmol) were added to 20 mL of N,N-dimethylacetamide and reacted at 100 ° C. overnight. The mixture was sampled on a plate for monitoring, and after the raw materials were completely reacted, it was treated to obtain 1.0 g of the product. 1H NMR (400 MHz, chloroform-d) δ 9.56 (s, 1H), 8.88 (s, 1H), 7.92 (d, J = 14.6Hz, 2H), 7.54 (s, 1H), 7.40 (d, J = 8.2 Hz, 1H), 7.32 (s, 1H), 7.15 (t, J = 7.6 Hz, 1H), 6.84 (s, 1H), 4.06 (s, 3H), 3.93 (s, 3H), 3.04 (t, J = 6.0 Hz, 1H), 2.88 (s, 3H), 2.72 (s, 3H), 2.44 (s, 2H), 2.38 (s, 3H), 1.28 (s, 4H).

[0048] Synthesis of compound 10: Compound 9 (1.0 g, 1.72 mmol) and palladium carbon (500 mg, wet palladium carbon 55%) were placed in 50 mL of methanol and reacted at room temperature for 4 hours in a hydrogen gas atmosphere. The mixture was sampled and monitored on a plate, and after the raw materials were completely reacted, 900 mg of the product was obtained.

[0049] Synthesis of Example 5: Compound 10 (900 mg, 1.63 mmol) and triethylamine (495 mg, 4.90 mmol) were dissolved in dichloromethane, and the temperature was lowered to 0°C in a nitrogen gas atmosphere. A dichloromethane solution of acrylic chloride (223 mg, 2.45 mmol) was then added dropwise, and the reaction was carried out for 2 hours while maintaining the temperature at 0°C. The mixture was sampled on a plate for monitoring, and after the raw materials had completely reacted, it was processed to obtain 200 mg of the product. [M+H] + :606.8.

[0050] With reference to Example 5, three deuterated compounds in Examples 6, 7, and 8 were synthesized, and the details are shown in Table 2 below. Table 2. Structures and Characterization of Compounds of Examples 6-8 [Table 2]

[0051] Example 9 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0052] The synthetic route of the compound of this example is as follows. [ka]

[0053] The synthesis process of the compound of this example is as follows. Synthesis of compound 3: Compound 1 (6.0 g, 13.30 mmol), compound 2 (5.03 g, 39.91 mmol) and N,N-diisopropylethylamine (3.43 g, 26.6 mmol) were added to 60 mL of N,N-dimethylacetamide and reacted at 80 ° C. overnight. The mixture was sampled on a plate for monitoring, and after the raw materials were completely reacted, it was treated to obtain 6.17 g of the product. 1 H NMR (400 MHz, chloroform-d)-δ 9.50 (s, 1H), 8.90 (s, 1H), 8.24 -7.99 (m, 1H), 7.80 (s, 1H), 7.67 (s, 1H), 7.38 (dt, J = 8.2, 1.0 Hz, 1H), 7.31-7.23 (m, 1H), 7.23-7.16 (m, 1H), 6.65 (s, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 3.70 (s, 3H), 2.95 (s, 3H), 2.89 (t, J = 6.6 Hz, 1H), 2.85-2.76 (m, 2H), 2.27 (m, 2H), 2.15 (s, 3H), 1.91 - 1.81 (m, 2H).

[0054] Synthesis of compound 4: Compound 3 (6.17 g, 11.08 mmol) and lithium hydroxide monohydrate (2.33 g, 55.39 mmol) were added to a mixed solvent of tetrahydrofuran / methanol / water with a volume ratio of 6:3:1, and reacted at 40° C. for 16 hours. The mixture was sampled on a plate for monitoring, and after the raw materials were completely reacted, it was treated to obtain 6.02 g of product.

[0055] Synthesis of compound 5: Compound 4 (6.02g, 11.08mmol), t-butyl carbazate (4.39g, 33.26mmol), N,N-diisopropylethylamine (8.58g, 66.48mmol) were added to 160 mL N,N-dimethylformamide and stirred for 15 min, after which 1H-benzotriazol-1-yloxytripyrrolidinohexafluorophosphate (6.90g, 13.30mmol) was added and reacted at 25°C for 16 hours. The mixture was sampled on a plate for monitoring, and after the raw materials had completely reacted, it was processed to obtain 1.67g of product.

[0056] Synthesis of compound 6: Compound 5 (1.67g, 2.54mmol) was dissolved in 15mL tetrahydrofuran, slowly added to 15mL of 4M hydrogen chloride in dioxane solution, and reacted at 40°C for 5 hours under nitrogen gas protection. After the raw materials were completely reacted, the reaction was monitored by liquid chromatography-mass spectrometry, and 1.20g of the product was obtained.

[0057] Synthesis of compound 7: Compound 6 (1.0g, 1.68mmol) and triethylamine (679 mg, 6.72mmol) were added to 20 mL of dichloromethane and stirred for 15 minutes. Trifluoroacetic anhydride (1.59g, 7.58mmol) was then added in portions and reacted at 40°C for 8 hours to complete the reaction. After treatment, 521mg of product was obtained.

[0058] Synthesis of compound 8: Compound 7 (521 mg, 0.82 mmol) and palladium carbon (156 mg, 10%) were dissolved in 20 mL of methanol, and reacted for 3 hours under stirring at 25° C. in a hydrogen gas atmosphere. The reaction was monitored by liquid chromatography-mass spectrometry. After the raw materials were completely reacted, the product was treated to obtain 412 g.

[0059] Synthesis of Example 9: Compound 8 (412 mg, 0.68 mmol) and triethylamine (206 mg, 2.04 mmol) were dissolved in 15 mL of dichloromethane, and the temperature was lowered to 0°C in a nitrogen gas atmosphere. A dichloromethane solution of acrylic chloride (93 mg, 1.02 mmol) was then added dropwise, and the mixture was allowed to react for 2 hours while maintaining the temperature at 0°C. The mixture was sampled and monitored on a plate, and after the raw materials had completely reacted, it was processed to obtain 122 mg of the product. [M+H] + :660.5; 1 H NMR (400 MHz, chloroform-d) δ 11.36 (m, 1H), 9.87 (m, 1H), 9.09 (m, 1H), 8.92 (s, 1H), 8.37 (m, 1H), 7.95 (m, 1H), 7.33 (m, 1H), 7.18 (m, 1H), 7.01 (m, 2H), 6.80 (m, 1H), 6.46 (d, J = 16.6 Hz, 1H), 5.74 (d, J = 10.2 Hz, 1H), 3.92 (d, J = 13.5 Hz, 6H), 3.29 (m, 2H), 3.04 (m, 1H), 2.78 (m, 8H), 2.21 (m, 2H).

[0060] With reference to Example 9, three deuterated compounds in Examples 10, 11, and 12 were synthesized, and the details are shown in Table 3 below. Table 3. Structures and Characterization of Compounds of Examples 10-12 [Table 3]

[0061] Example 13 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0062] The synthetic route of the compound of this example is as follows. [ka]

[0063] Synthesis of compound 2: In a 500mL single-neck flask, compound 1 (10.00g, 62.88mmol) was added and completely dissolved in 100mL tetrahydrofuran. Cesium carbonate (20.50g, 62.88mmol) and trifluoroethanol (6.29g, 62.87mmol) were added, and after the addition was completed, the flask was protected with nitrogen gas. The mixture was sampled on a plate for monitoring, and reacted at 23°C for 6 hours, resulting in an almost complete reaction. After processing, 14.35g of product was obtained, with a yield of 95.5%. 1 H NMR (400 MHz, chloroform-d) δ 8.01 (dd, J = 9.1, 5.8 Hz, 1H), 7.09 - 6.72 (m, 2H), 4.50 (q, J = 7.8 Hz, 2H).

[0064] Synthesis of compound 3: Compound 2 (14.00g, 58.57mmol) was dissolved in 60mL ethanol, 15mL water, ammonium chloride (9.60g, 179.44mmol), reduced iron powder (20.00g, 357.14mmol) were added, and after the addition was completed, the temperature was raised to 80°C and reacted overnight. The reaction was monitored by sampling on a plate, and the reaction was completed and treated to obtain 10.50g of product, with a yield of 85.78%. 1 H NMR (400 MHz, chloroform-d) δ 6.89 - 6.34 (m, 3H), 4.34 (q, J = 8.0 Hz, 2H), 3.84 - 3.27 (m, 2H).

[0065] Synthesis of compound 4: In a 250mL three-neck flask, compound 3 (10.00g, 47.83mmol) was added, and 50mL concentrated sulfuric acid was added while stirring. The temperature was lowered to 0°C, and potassium nitrate solid (6.10g, 60.34mmol) was added in portions. After the addition was completed, the mixture was protected with nitrogen gas. After reacting at room temperature for 4 hours, the mixture was sampled on a plate and treated after the reaction was completed. 8.26g of the product was obtained, with a yield of 67.98%. 1 H NMR (400 MHz, chloroform-d) δ 7.44 (d, J = 7.2 Hz, 1H), 6.66 (d, J = 11.5 Hz, 1H), 4.46 (q, J = 7.7 Hz, 2H), 4.01 (m, 2H).

[0066] Synthesis of compound 6: Compound 4 (2.00 g, 6.08 mmol) and compound 5 (2.00 g, 7.87 mmol) were added to 50 mL of acetonitrile, and p-toluenesulfonic acid monohydrate (0.81 g, 4.26 mmol) was added and stirred at 80 ° C overnight. The next day, the raw material was almost completely reacted by sampling on a plate and monitoring. The temperature was lowered, and a solid precipitated from the reaction liquid. It was filtered, and the cake was dried by rotation, and then methanol was added and mixed to obtain 1.67 g of product. 1 H NMR (400 MHz, chloroform-d) δ 9.89 (s, 1H), 9.33 (d, J = 8.0 Hz, 1H), 8.77 (s, 1H), 8.29 (s, 1H), 7.67 (d, J = 8.0 Hz, 1H), 7.39 (m, 1H), 7.33 (m, 1H), 7.19 (m, 1H), 6.89 (d, J = 11.3 Hz, 1H), 5.14 (p, J = 6.2 Hz, 1H), 4.59 (q, J = 7.7 Hz, 2H), 3.95 (s, 3H), 1.22 (d, J = 6.2Hz, 6H).

[0067] Synthesis of compound 8: Compound 6 (1.52 g, 2.78 mmol), compound 7 (1.05 g, 8.33 mmol) and N,N-diisopropylethylamine (1.05 g, 8.14 mmol) were added to 25 mL of N,N-dimethylacetamide and reacted at 90 ° C overnight. The mixture was sampled on a plate for monitoring, and after the raw materials were completely reacted, it was treated to obtain 1.16 g of product.

[0068] Synthesis of compound 9: Compound 8 (0.59 g, 0.90 mmol) and palladium carbon (400 mg, wet palladium carbon 55%) were placed in a mixed solvent of 20 mL methanol and 20 mL ethyl acetate, and reacted at room temperature for 3 hours in a hydrogen gas atmosphere. The mixture was sampled and monitored on a plate, and after the raw materials were completely reacted, 520 mg of the product was obtained.

[0069] Synthesis of Example 13: Compound 9 (520 mg, 0.83 mmol) and triethylamine (253 mg, 2.50 mmol) were dissolved in dichloromethane, and the temperature was lowered to 0°C under a nitrogen gas atmosphere. A dichloromethane solution of acrylic chloride (114 mg, 1.25 mmol) was then added dropwise, and the mixture was allowed to react for 2 hours while maintaining the temperature at 0°C. The mixture was sampled on a plate for monitoring, and after the raw materials had completely reacted, it was processed to obtain 170 mg of the product. [M+H] + :606.8. 1H NMR (400 MHz, chloroform-d) δ 10.63 (s, 1H), 10.11 (s, 1H), 8.82 (s, 1H), 8.16 (s, 1H), 7.66 (m, 2H), 7.29 (d, J = 8.1 Hz, 1H), 7.22 - 7.16 (m, 1H), 7.15 - 7.09 (m, 1H), 6.80 (s, 1H), 6.37 (dd, J = 16.9, 2.0 Hz, 1H), 5.68 (dd, J = 10.1, 2.0 Hz, 1H), 5.00 (p, J = 6.2 Hz, 1H), 4.44 (q, J = 8.2 Hz, 2H), 3.85 (s, 3H), 3.27 - 2.99 (m, 2H), 2.87 - 2.51 (m, 6H), 2.11 (m, 2H), 1.18 - 0.91 (m, 6H).

[0070] Example 14 was synthesized with reference to Example 13, and the details are shown in Table 4 below. Table 4. Structure and characterization of the compound of Example 14 [Table 4]

[0071] Example 15 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0072] The synthetic route of the compound of this example is as follows. [ka]

[0073] Synthesis of Example 15: In a 50mL single-neck flask, compound 2 (112mg, 1.24mmol), 20ml of dichloromethane, and oxalyl chloride (142mg, 1.12mmol) were placed and stirred for 2 hours under nitrogen gas protection. The temperature was lowered to 0°C, and a dichloromethane solution of compound 1 (438mg, 78.92mmol) and triethylamine (152mg, 1.50mmol) were added dropwise, and the reaction was carried out for 1 hour while maintaining the temperature at 0°C. The mixture was sampled and monitored on a plate, and after the raw materials were completely reacted, the mixture was treated, and 160mg of product was finally obtained. [M+H] + :628.8; 1 H NMR (400 MHz, chloroform-d)δ 9.53 (m, 1H), 9.12 (m, 1H), 8.89 (s, 1H), 8.35 (m, 1H), 7.92 (s, 1H), 7.69 (s, 1H), 7.35 (d, J = 8.1 Hz, 1H), 7.23 (t, J = 7.5 Hz, 1H), 7.15 (t, J = 7.5 Hz, 1H), 6.84 (s, 1H), 5.84 (dd, J = 48.2, 3.4 Hz, 1H), 5.28 (dd, J = 15.5, 3.4 Hz, 1H), 5.04 (p, J = 6.2 Hz, 1H), 3.92 (s, 6H), 3.49 (m, 2H), 2.64 (m, 9H), 2.19(m, 2H), 1.09 (m, 6H).

[0074] Example 16 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0075] The synthetic route of the compound of this example is as follows. [ka]

[0076] Synthesis of compound 3: In a 100 mL single-neck flask, compound 1 (313 mg, 1 mmol) was added, 5 mL of acetonitrile was added, and the crude product of compound 2 (441 mg) was further added. After the addition was completed, the flask was protected with nitrogen gas. The mixture was sampled on a plate for monitoring, and reacted overnight at 80°C, whereupon the reaction was almost complete. After processing, 300 mg of the product was obtained.

[0077] Synthesis of compound 4: Compound 3 (1.83 g, 4.54 mmol) was dissolved in 60 mL of methanol, and 1.2 mL of concentrated hydrochloric acid was added. After the addition was completed, the temperature was raised to 60° C. and the reaction was allowed to proceed overnight. The reaction was monitored by sampling on a plate, and the reaction was completed. After treatment, 0.89 g of the product was obtained. 1 H NMR (400 MHz, chloroform-d) δ 7.61 (s, 1H), 4.80 (q, J = 8.4 Hz, 2H), 3.57 (m, 2H), 3.10 (s, 3H), 3.02 (t, J = 6.9 Hz, 1H), 2.59 - 2.33 (m, 4H), 2.06 (s, 3H), 1.76 - 1.51 (m, 2H).

[0078] Synthesis of compound 6: In a 100 mL single-neck flask, compound 4 (73.8 mg, 0.21 mmol), compound 5 (56 mg, 0.17 mmol), p-toluenesulfonic acid (48 mg, 0.25 mmol), and 5 mL of dioxane were added, and the mixture was stirred overnight at 80° C. and protected with nitrogen gas. The mixture was sampled on a plate for monitoring, and after the reaction was completed, it was treated to obtain 50 mg of the product. 1H NMR (400 MHz, chloroform-d) δ 9.76 (s, 1H), 8.90 (s, 1H), 7.94 (s, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.56 (s, 1H), 7.39 (d, J = 8.2 Hz, 1H), 7.33 - 7.27 (m, 1H), 7.24 - 7.17 (m, 1H), 5.10 (p, J = 6.2 Hz, 1H), 4.94 (s, 2H), 3.93 (s, 3H), 3.78 (m, 2H), 3.18 (t, J = 6.5 Hz, 1H), 3.01 (m, 3H), 2.57 (m, 5H), 2.31 (m, 2H), 1.16 (d, J = 6.2 Hz, 6H).

[0079] Synthesis of compound 7: Compound 6 (98 mg, 0.15 mmol) and palladium carbon (80 mg, wet palladium carbon 55%) were placed in a mixed solvent of 3 mL methanol and 2 mL ethyl acetate, and reacted at room temperature for 2 hours in a hydrogen gas atmosphere. The mixture was sampled on a plate for monitoring, and after the raw materials were completely reacted, 98 mg of the product was obtained.

[0080] Synthesis of Example 16: Compound 8 (300 mg, 0.48 mmol) and triethylamine (150 mg, 1.48 mmol) were dissolved in dichloromethane, and the temperature was lowered to 0°C under a nitrogen gas atmosphere. A dichloromethane solution of acrylic chloride (70 mg, 0.77 mmol) was then added dropwise, and the mixture was allowed to react for 2 hours while maintaining the temperature at 0°C. The mixture was sampled on a plate for monitoring, and after the raw materials had completely reacted, it was processed to obtain 118 mg of the product. [M+H] + :688.8. 1H NMR (400 MHz, chloroform-d) δ 10.39 (s, 1H), 9.97 (s, 1H), 8.89 (s, 1H), 8.59 (m, 1H), 7.59 (m, 2H), 7.33 (m, 1H), 7.23 (m, 1H), 7.14 (m, 1H), 6.45 (m, 1H), 6.31 (m, 1H), 5.72 (d, J = 10.0 Hz, 1H), 5.01 (p, J = 6.5 Hz, 1H), 4.83 (q, J = 8.5 Hz, 2H), 3.96 (s, 3H), 3.14 - 2.88 (m, 2H), 2.85 - 2.52 (m, 6H), 2.33 (s, 3H), 1.69 (m, 2H), 1.04 (d, J = 6.3 Hz, 6H).

[0081] Example 17 The compound used as the kinase inhibitor in this example has the following structural formula: [ka]

[0082] The synthetic route of the compound of this example is as follows. [ka]

[0083] Synthesis of compound 2: In a 100mL three-neck flask, compound 1 (153mg, 0.38mmol) was added, 5mL of tetrahydrofuran was added, and 4-dimethylaminopyridine (17.8mg) was further added. After the addition was completed, di-t-butyl dicarbonate (1.06g, 4.86mmol) was added dropwise under the protection of nitrogen gas. The mixture was sampled on a plate for monitoring, and reacted at 80°C for 2 hours until the reaction was almost complete. After processing, 104mg of product was obtained.

[0084] Synthesis of compound 3: Compound 2 (104 mg, 0.21 mmol) was dissolved in 2 ml of methanol, and 0.05 mL of 5.4 mol / L sodium methoxide in methanol was added. After the addition was completed, the reaction was allowed to proceed for 30 minutes. The reaction was monitored by sampling on a plate, and the reaction was completed. After processing, 30 mg of the product was obtained.

[0085] Synthesis of compound 4: Compound 3 (30 mg, 0.065 mmol) and palladium carbon (15 mg, wet palladium carbon 55%) were placed in a mixed solvent of 1 mL methanol and 1 mL ethyl acetate, and reacted at room temperature for 2 hours in a hydrogen gas atmosphere. The mixture was sampled and monitored on a plate, and after the raw materials had completely reacted, the product was treated to obtain 30 mg.

[0086] Synthesis of compound 5: Compound 4 (30 mg, 0.069 mmol) and triethylamine (13 mg, 0.14 mmol) were dissolved in dichloromethane, and the temperature was lowered to 0° C. in a nitrogen gas atmosphere. A dichloromethane solution of acrylic chloride (7 mg, 0.077 mmol) was then added dropwise, and the mixture was allowed to react for 2 hours while maintaining the temperature at 0° C. Samples were placed on a plate for monitoring, and after the raw materials had completely reacted, the mixture was processed to obtain 12 mg of product.

[0087] Synthesis of Example 17: Compound 5 (12 mg, 0.024 mmol), Compound 6 (6 mg, 0.018 mmol), and p-toluenesulfonic acid (8 mg, 0.042 mmol) were dissolved in 1 mL of N-methylpyrrolidone and 2 mL of ethylene glycol monomethyl ether, and the mixture was heated to 100° C. in a nitrogen gas atmosphere and reacted for 4 hours. The mixture was sampled on a plate for monitoring, and after the raw materials had completely reacted, it was processed to obtain 3 mg of the product. [M+H] + :675.8. 1H NMR (400 MHz, chloroform-d) δ 10.42 (s, 0H), 9.97 (s, 0H), 8.91 (s, 0H), 8.70 (m, 1H), 7.68 (s, 1H), 7.33 (d, J = 8.2 Hz, 0H), 7.18 (m, 1H), 6.99 (m, 1H), 6.45 (m, 1H), 6.33 (m, 1H), 5.73 (m, 1H), 5.35 (m, 1H), 4.85 (m, 2H), 3.99 (s, 3H), 3.05 (m, 2H), 2.78 (m, 1H), 2.62(m,2H), 2.35 (s, 3H), 2.22 (s, 3H), 2.10 - 1.95 (m, 2H).

[0088] 2. Biological testing and evaluation of compounds 1. Testing the inhibitory effect of compounds on cell proliferation 1.1 Experiments to test the cytostatic activity of EGFR and HER2 exon 20 insertion mutations: The engineered cells Ba / F3-FL-EGFR-V769-D770 ins ASV, Ba / F3-FL-EGFR-D770-N771 ins SVD, Ba / F3-FL-EGFR-H773-V774 ins NPH, Ba / F3-FL-EGFR- A763-Y764 ins FQEA, Ba / F3-HER2-A775-G776 ins YVMA selected in the experiment were provided by Hefei Zhongke Pu Ruisheng Biomedical Technology Co., Ltd. and were validated before being used in this study. A 20× stock solution of test compound was prepared for use, diluted in 3-fold gradient dilutions at 9 concentrations starting from 1 μM.

[0089] The cell suspension in logarithmic phase was taken and inoculated into a 96-well white cell culture plate, with a volume of 95 μL (2000 cells / well) in each well. 5 μL of 20× test compound was taken and added to the culture plate containing the above 95 μL cell suspension with reference to the plate inoculation diagram, and mixed evenly. Incubated in an incubator at 37°C and 5% CO2 for 72 hours. The proliferation inhibition activity of the compounds was measured by CellTiter-Glo method. The corresponding fluorescence value RLU of each well was obtained from the readings of SpectraMax Paradigm. The data of the cell proliferation inhibition rate (Inhibition Rate) was processed by the following formula: Inhibition Rate (Inh%)=100-(RLU compound-RLU blank) / (RLU control-RLU blank)*100%. After calculating the cell proliferation activity corresponding to different concentrations of the compound in EXCEL, the cell proliferation activity curve was drawn in GraphPad Prism software to obtain the IC 50 The values ​​were calculated. Table 5 shows the names and structures of the control products.

[0090] Table 5. Name and structure of control products [Table 5]

[0091] The experimental results in Tables 6, 7 and 8 show that IC 50 There are three types depending on the value, where A≦30nM, 30nM<B≦100nM、C> It is 100nM. Table 6. Test results of the cytostatic activity of different compounds against EGFR exon 20 insertion mutations [Table 6]

[0092] Table 7. Test results of the cytostatic activity of different compounds against HER2 exon 20 insertion mutations [Table 7]

[0093] From Table 6, it can be seen that all the compounds in the examples have good inhibitory ability against EGFR exon 20 insertion mutation. It is better than DZD9008 and corresponds to the activity of TAK-788. From Table 7, it can be seen that the cell activity of Example 5 in HER2 exon 20 insertion mutation is equivalent to TAK-788 and better than DZD9008.

[0094] 1.2 Experiments to test the inhibitory activity of compounds against cell proliferation in cell lines NCI-H1975 and PC9: The cell lines NCI-H1975 and PC9 selected in the experiment were provided by China American Crown Biotechnology (Beijing) Co., Ltd. and were validated before being used in this study. A 10× stock solution of test compound was prepared for use, diluted in 4-fold gradient dilutions at nine concentrations starting from 10 μM.

[0095] The logarithmic phase cell suspension was taken and inoculated into a 96-well culture plate, with the volume of each well being 90 μL (2000 cells / well). 10 μL of 10× test compound was taken and placed in each culture plate, referring to the plate inoculation diagram, and mixed evenly. Incubated at 37℃ in a 5% CO2 incubator. The proliferation inhibitory activity of the compounds was measured by the CellTiter-Glo method. The inhibition rate data was obtained by creating a cell vitality curve with GraphPad Prism 8.0 software and calculating the IC 50 The values ​​were calculated and the results are shown in Table 8.

[0096] Table 8. Test results of the cytostatic activity of different compounds on related mutations [Table 8]

[0097] From Table 8, it can be seen that the compounds of the present invention have excellent inhibitory effects on cell proliferation of cell lines NCI-H975 and PC9, and are better than DZD9008.

[0098] 2. In vivo efficacy test of compounds 2.1 Pharmacology study of the LU0387 PDX lung cancer model The lung cancer LU0387PDX model is a commonly used xenograft tumor model in the Coronary Artery Disease Center, and the experimental conditions of this model are as follows: After euthanizing the tumor-bearing mice, the tumor mass was removed under aseptic conditions, washed, and blood stains, connective tissue and necrotic parts were removed, and the tumor was divided into 2×2×2mm 3 The mice were inoculated subcutaneously into the right dorsal region of the experimental mice using a disinfected inoculation needle. The tumor growth was periodically monitored and the tumors were grown to an average volume of 100-200 mm. 3 When the tumors grew to 100 mm, they were randomly divided into groups according to tumor size and mouse weight for administration. Before administration began, all animals were weighed, and tumor diameters were measured with a caliper. To ensure that tumor volumes were comparable among different groups, mice were randomly divided into groups according to tumor volume. Each group contained three mice, and the mice were orally administered once a day, and the inoculation site was observed after administration. Tumor volumes and mouse weights were measured twice a week.

[0099] The experimental design and results are shown in Table 9, where PO is oral administration, QD is once a day, and TGI (tumor volume inhibition rate) = (1-tumor weight of treatment group / tumor weight of control group) * 100%.

[0100] Table 9. LU0387 model design and results [Table 9]

[0101] In the experiment, the tumor volume changed depending on the number of days of administration, and the results are shown in FIG. From Figure 1, it can be seen that the efficacy of Example 1 was better than TAK-788 and DZD9008 at the same dose in the LU0387 model. The efficacy of Example 5 was similar to TAK-788. With the increase in the dose of Example 1, the efficacy was obviously enhanced.

[0102] 2.2 In vivo drug efficacy study on Ba / F3-EGFR-D770_N771 ins SVD engineered cell line subcutaneously transplanted tumor model The cell line Ba / F3 EGFR D770_N771 ins SVD was cultured in RPMI1640 + 10% fetal bovine serum + 1% double antibiotics at 37°C, 5% CO2 and passaged 2-3 times per week. Cells were harvested when they reached 80-90% confluence and number required. 0.2ml (1*106 cells) of cells were inoculated subcutaneously into the right back of female nude mice aged 6-8 weeks and weighing 18-22g. The average tumor volume was approximately 150-200 mm 3 When the tumor volume reached 100 mg / kg, the mice were divided into groups and administered the drug. Before the start of administration, all animals were weighed and tumor diameters were measured with a caliper. The mice were randomly divided into groups according to their tumor volume so that the tumor volumes among different groups were equivalent. Each group contained three mice, and the mice were orally administered the drug once a day. After administration, the inoculation site was observed. The tumor volume and mouse weight were measured twice a week.

[0103] Table 10. Grouping of Ba / F3 EGFR D770_N771 ins SVD model [Table 10]

[0104] Examples 18 and 40 in the table are compounds having a trans tripentamine structure in Patent WO2021180238. The names and structures are shown in Table 11.

[0105] Table 11. Names and structures of compounds with trans structure [Table 11]

[0106] In the experiment, the tumor volume changed depending on the number of days of administration, and the results are shown in FIG. From FIG. 2, it can be seen that in the Ba / F3-EGFR-D770_N771 ins SVD cell transplantation model, at the same dosage, the internal pharmacological efficacy of both Example 1 and Example 5 is far better than that of the compound having a trans-tripentamine structure in Patent WO2021180238, and the tumor inhibition rate is improved by more than 1.6 times.

[0107] As mentioned above, the compounds in this patent have good inhibitory effects on EGFR or HER2 exon 20 insertion mutation, and also have good inhibitory effects on cell proliferation in cell lines NCI-H1975 and PC9. For one or more combinations of EGFR exon 20 insertion mutation, HER2 exon 20 insertion mutation, EGFR exon 19 deletion, EGFR exon 20 point mutation, and EGFR exon 21 point mutation, these drugs have good therapeutic effects on related diseases.

[0108] 3. Structural confirmation of Example 1 and research into the free base crystal form [ka]

[0109] 1. Structure confirmation 214 mg of the compound of Example 1 was taken, 0.5 mL of dichloromethane and 0.5 mL of acetonitrile were added, and the whole was dissolved and filtered. The filtrate was placed in a sample bottle with a sealing membrane, and after opening a hole, it was slowly evaporated in a draft chamber to obtain granular crystals (free base crystal form I). The structure was confirmed by single crystal diffraction, and the result is shown in Figure 3, and the configuration of the compound was confirmed.

[0110] 2. Study of the Free Base Crystalline Form of Example 1 Polymorph screening was performed on the compound obtained in Example 1 to search for potential crystal forms so as to find a good crystal form suitable for further development. The compound was detected using the synthesized Example 1 as the starting material, and was found to have good crystallinity and an anhydrous crystal form, so it was named free base crystal form I. Then, an amorphous free base compound was produced by dry grinding. Using the above two types as starting materials, a crystal form screening experiment was performed.

[0111] 2.1 Preparation and characterization of starting materials 2.1.1 Characterization of the Free Base Crystalline Form I of the Compound of Example 1 The raw material of the free base crystalline form I of the compound of Example 1 was fully characterized. Polarized light microscopy (PLM) showed that the raw material was irregularly shaped crystals with good crystallinity. Figure 4 shows the XRPD spectrum of the free base crystalline form I, and Figure 5 shows the overlay of DSC and TGA of the crystalline form. The differential scanning calorimetry (DSC) curve shows one endothermic peak at 230°C, which becomes a melting peak. The thermogravimetric analysis (TGA) curve shows that there is no obvious weight loss before decomposition, which indicates that it is an anhydrous crystalline form.

[0112] Table 12. Powder X-ray diffraction peak data for free base crystalline form I [Table 12]

[0113] 2.1.2 Solubility Test of Free Base Crystalline Form I of Example 1 The solubility of the free base crystalline form I of compound Example 1 at room temperature was measured by visual measurement in 18 different solvents. The results are shown in Table 13. The free base crystalline form I was highly soluble only in dichloroethane and dichloromethane, and was poorly soluble in most of the solvents measured.

[0114] Table 13. Results of visual measurement of solubility of free base crystalline form I of Example 1 [Table 13]

[0115] 2.1.3 Preparation and Characterization of Amorphous Feedstock 736 mg of the compound of Example 1 (free base crystalline form I) was dry-polished for 5 hours to produce 669.86 mg of amorphous product. XRPD detection was performed and the result is shown in FIG.

[0116] 2.2 Methods and parameters of physical and chemical detection devices used in characterization 2.2.1 X-ray powder diffraction (XRPD) XRPD diffraction spectra were obtained using a Bruker D2 Phaser model. The test samples were placed on a smooth, background-free silicon sheet. The measurement parameters are shown in Table 14.

[0117] Table 14 XRPD method parameters [Table 14]

[0118] 2.2.2 Polarized Light Microscopy (PLM) PLM analysis was performed using an OPTEC optical microscope BK-Pol. A small amount of the sample was taken, placed on a slide glass, dispersed with a drop of silicone oil, covered with a cover glass, and observed under the microscope.

[0119] 2.2.3 Differential Scanning Calorimetry (DSC) The DSC curve was obtained by a TA instrument, DSC 250. The measurement method for the DSC 250 instrument was to precisely measure an appropriate amount of sample, place it in an aluminum crucible (a hole was drilled in the sample), and raise the temperature from 25°C to a final temperature of 300°C at a heating rate of 10°C / min, and blow with nitrogen gas at a flow rate of 50 mL / min.

[0120] 2.2.4 Thermogravimetric analysis (TGA) TGA data were obtained by a TA instrument, model TGA 550. An appropriate amount of sample was placed in the aluminum crucible with the surface removed, and the temperature was increased from room temperature to 300 °C at a heating rate of 10 °C / min, the balance chamber was blown with nitrogen gas at 40 mL / min, and the sample chamber was blown with nitrogen gas at 25 mL / min.

[0121] 2.3 Study of free base crystalline polymorphism of the compound of Example 1 They were mainly prepared by methods such as suspension crystallization, anti-solvent precipitation, high and low temperature circulation and evaporative crystallization. Suspension crystallization was carried out at 25°C and 50°C using the free base crystalline form I and the amorphous crystalline form as starting materials, respectively, in a single solvent (solvent selected from dichloromethane, 1,4-dioxane, dichloroethane, methyl-t-butyl ether, N-methylpyrrolidone, ethyl acetate, acetone, methanol, dimethylsulfoxide, isopropyl acetate, butanone, cyclohexane, tetrahydrofuran, water, acetonitrile, isopropanol, ethanol, and n-heptane) or a mixed solvent selected from the group consisting of dichloromethane, 1,4-dioxane, dichloroethane, methyl-t-butyl ether, N-methylpyrrolidone, ethyl acetate, acetone, methanol, dimethylsulfoxide, isopropyl acetate, butanone, cyclohexane, tetrahydrofuran, water, acetonitrile, isopropanol, ethanol, and n-heptane, based on the results of the visual solubility test of the compound in Example 1. A total of two crystalline forms were obtained, namely, free base crystalline form I and free base crystalline form II.

[0122] In the poor solvent precipitation, dichloromethane and dichloroethane were selected as good solvents based on the results of the visual measurement of the solubility of the compound in Example 1, and different poor solvents were added to each of them under stirring conditions at room temperature (up to 25°C) to produce the crystal forms. A total of two crystal forms were obtained, free base crystal form I and free base crystal form III.

[0123] In the high and low temperature circulation, the compound was mixed with a solvent (the solvent was selected from dichloromethane, 1,4-dioxane, dichloroethane, methyl-t-butyl ether, N-methylpyrrolidone, ethyl acetate, acetone, methanol, dimethyl sulfoxide, isopropyl acetate, butanone, cyclohexane, tetrahydrofuran, water, acetonitrile, isopropanol, ethanol, and n-heptane) and stirred under temperature cycling conditions of 50°C to 5°C to produce the crystal form. A total of two crystal forms were obtained: free base crystal form I and free base crystal form II.

[0124] Evaporative crystallization was carried out by adding a solvent to the compound, filtering the prepared solution, dropping the filtrate into a sample bottle, and slowly evaporating it in a fume hood after puncturing the sealing membrane covering the opening. Two crystal forms were obtained: free base crystal form I and free base crystal form IV. After heating free base crystal form IV to 150 °C to remove the solvent, a new crystal form was produced, named free base crystal form V.

[0125] 2.3.1 Preparation and Characterization of Free Base Form I The free base crystalline form I was obtained in most solvents, and the compound obtained in Example 1 is the free base crystalline form I. The characterization results are shown in Figures 4 and 5.

[0126] 2.3.2 Preparation and Characterization of Free Base Crystalline Form II Free base crystalline form II is obtained in a solvent system containing a part of ethanol, and free base crystalline form II is obtained by suspension crystallization of amorphous compound in a single solvent, ethanol. The characterization results are shown in Figures 7 and 8, the DSC curve of free base crystalline form II has two endothermic peaks at 124°C and 230°C, the TGA curve has a weight loss of 4.48% at 65-150°C, and free base crystalline form II is obtained in a solvent system of ethanol. From the above, it is inferred that free base crystalline form II is a solvate of ethanol. The endothermic peak at 124°C in the DSC curve is the desolvation peak.

[0127] Table 15. Powder X-ray diffraction peak data for free base crystalline form II [Table 15]

[0128] 2.3.3 Preparation and Characterization of Free Base Crystalline Form III Free base crystalline form III is obtained in a solvent system containing some isopropanol, and when the crystalline form is prepared by anti-solvent precipitation, free base crystalline form III is found in dichloromethane / isopropanol or dichloroethane / isopropanol systems. The characterization results are shown in Figures 9 and 10, the DSC curve of free base crystalline form III has two endothermic peaks at 121°C and 230°C, the TGA curve shows a weight loss of 9.57% at 75-145°C, and after heating to 150°C to remove the solvent, the free base crystalline form III is transformed to free base crystalline form I in XRPD. Free base crystalline form III is obtained in a solvent system of isopropanol, and from the above, it is inferred that free base crystalline form III is a solvate of isopropanol, and the endothermic peak at 121°C in the DSC curve is the desolvation peak.

[0129] Table 16. Powder X-ray diffraction peak data for free base crystalline form III [Table 16]

[0130] 2.3.4 Preparation and Characterization of Free Base Form IV Free base crystalline form IV is obtained in a solvent system containing some dichloroethane. 92.28mg of raw material is taken and placed in a sample bottle, and 5.0mL of dichloroethane is added to dissolve it. The sample bottle is covered with a sealing film and pierced, and then slowly evaporated in a fume hood, thus preparing crystalline form IV by evaporation crystallization. Its characterization results are shown in Figures 11 and 12. The DSC curve of free base crystalline form IV has two endothermic peaks at 108℃ and 231℃, and the TGA curve shows a weight loss of 10.41% at 25-150℃.1 From the H NMR results, it is estimated that the sample contains 4.27% water, since only 6.14% of dichloroethane solvent remains. From the above, it is estimated that the free base crystalline form IV is a solvate of a mixture of dichloroethane and water.

[0131] Table 17. Powder X-ray diffraction peak data for free base crystalline form IV [Table 17]

[0132] 2.3.5 Preparation and Characterization of Free Base Form V After heating the free base crystalline form IV to 150°C to remove the solvent, it was characterized by XRPD, which showed that it had become a new crystalline form, which was named free base crystalline form V. The characterization results are shown in Figures 13 and 14. The DSC curve of free base crystalline form V has an endothermic peak at 231°C, and the TGA curve shows no obvious weight loss before decomposition. From the above, it is inferred that free base crystalline form V is an anhydrous crystalline form.

[0133] Table 18. Powder X-ray diffraction peak data for free base crystalline form V [Table 18]

[0134] 3. Evaluation of the crystalline form of the free base Form I In the screening process, only two anhydrous crystal forms, free base crystal form I and free base crystal form V, were found. Saturated solutions were prepared using two solvents, acetone and acetonitrile, respectively, and competitive suspension was performed on both at 25℃ or 50℃. As a result, it was inferred that free base crystal form I is the thermodynamically stable crystal form of the compound. And free base crystal form V was not obtained directly in the screening experiment, but only by desolvation of free base crystal form IV. Therefore, the crystal form evaluation was performed only on free base crystal form I, including studies on dry grinding, wet grinding, tableting (30 MPa), stability and hygroscopicity.

[0135] The grinding experiment showed that the free base crystalline form I became amorphous after 5 min of dry grinding, there was no obvious change in crystallinity after 5 min of wet grinding with water, and the crystallinity slightly decreased after 5 min of wet grinding with ethanol. In the tableting experiment, the crystallinity decreased slightly when the pressure was 30 MPa.

[0136] In the stability experiment, the free base crystalline form I was exposed to 80℃ for 3 days, 60℃ for 7 days, 25℃ / 60%RH for 7 days, 40℃ / 75%RH for 7 days, and 25℃ / 90±5%RH for 7 days, and then samples were taken for XRPD detection. The results showed that the free base crystalline form I had no obvious changes, and the liquid phase detection results of the drug also had no changes.

[0137] A dynamic vapor sorption (DVS) test was performed on the free base crystalline form I, and the water weight gain at 80% RH was 0.66%, indicating that the free base crystalline form I is slightly hygroscopic.

[0138] As described above, the spatial configuration of this type of compound represented by formula 1 was identified, and in particular the spatial structure of Example 1 was identified. In the crystal form screening study of the compound of Example 1, a total of two anhydrous crystal forms (free base crystal form I and free base crystal form V) and three solvates (free base crystal form II, free base crystal form III and free base crystal form IV) were found. In the experiment, it was found that free base crystal form I is a relatively stable anhydrous crystal form, has stable solid properties, and is slightly hygroscopic, which is useful for subsequent drug development.

Claims

1. A compound for use as a kinase inhibitor, characterized in that the compound is represented by formula 1, or a deuterated derivative thereof, or a pharma- ceutically acceptable salt, solvate or prodrug thereof. 【Chemistry 1】 (In formula 1, X is selected from CH and N. R １ teeth 【Chemistry 2】 Selected from R ５ is H, a C1-C3 alkyl group, or a C1-C3 fluoroalkyl group. R ２０ , R ２１ , R ２２ are each independently selected from a methyl group or a deuterated methyl group. R ３ is selected from a C1-C3 alkyl group and a C1-C3 haloalkyl group. R ４０ , R ４１ , R ４２ are each independently selected from H, D, and F.

2. The compound used as a kinase inhibitor according to claim 1, characterized in that it is a compound represented by formula 2, or a deuterated derivative thereof, or a pharma- ceutically acceptable salt, solvate or prodrug thereof. 【Chemistry 3】

3. In formula 2, X is selected from CH and N; R ３ Ha-CH ３ , -CH ２ CH ３ , -CH ２ CF ３ Selected from R ４０ , R ４１ are both H and R ４２ The compound used as a kinase inhibitor according to claim 2, wherein is selected from H or F.

4. The compound used as a kinase inhibitor according to claim 1, characterized in that it is a compound represented by formula 3, or a deuterated derivative thereof, or a pharma- ceutically acceptable salt, solvate or prodrug thereof. 【Chemistry 4】

5. In formula 3, X is selected from CH and N; R ３ Ha-CH ３ , -CH ２ CH ３ , -CH ２ CF ３ So, R ４０ , R ４１ are both H and R ４２ is selected from H or F; R ５ Ha-CH ３ , -CF ３ The compound used as a kinase inhibitor according to claim 4, which is selected from the following:

6. The compound represented by formula 1 is as follows: 【Chemistry 5】 The compound for use as a kinase inhibitor according to claim 1, characterized in that it is

7. The compound used as a kinase inhibitor according to any one of claims 1 to 6, wherein the compound is in a crystalline form, an amorphous form or a solvate, and the solvent contained in the solvate is a non-aqueous solvent or a mixed solvent consisting of a non-aqueous solvent and water.

8. A compound having the structure represented by formula A, 【Chemistry 6】 The compound for use as a kinase inhibitor according to claim 7, characterized in that the crystalline form is free base crystalline form I, and in the powder X-ray diffraction spectrum of said crystalline form, there are characteristic diffraction peaks at 2θ of 9.76°±0.2°, 10.45°±0.2°, 16.54°±0.2°, 18.66°±0.2°, 20.07°±0.2°, and 25.90°±0.2°.

9. The powder X-ray diffraction spectrum of the free base crystalline Form I has 2θ values ​​of 9.07°±0.2°, 9.76°±0.2°, 10.45°±0.2°, 11.53°±0.2°, 11.80°±0.2°, 12.91°±0.2°, 13.79°±0.2°, 14.67°±0.2°, 15.08°±0.2°, 15.63°±0.2°, A compound for use as a kinase inhibitor as described in claim 8, characterized in that it has characteristic diffraction peaks at 16.54°±0.2°, 17.50°±0.2°, 18.66°±0.2°, 20.07°±0.2°, 21.10°±0.2°, 23.29°±0.2°, 24.16°±0.2°, and 25.90°±0.2°.

10. Use of a compound used as a kinase inhibitor according to any one of claims 1 to 9 in the manufacture of a medicament for the treatment of diseases associated with EGFR mutations and / or HER2 mutations.

11. The use according to claim 10, characterized in that the EGFR mutation and / or HER2 mutation comprises one or a combination of two or more of the following: EGFR exon 20 insertion mutation, HER2 exon 20 insertion mutation, EGFR exon 19 deletion, EGFR exon 21 point mutation, EGFR exon 20 point mutation.

12. The use according to claim 11, characterized in that the EGFR and / or HER2 mutations are selected from the EGFR Del 19 / T790M / C797S mutation, the EGFR L858R / T790M / C797S mutation.

13. The use according to claim 10, 11 or 12, characterized in that the disease is a cancer caused by said EGFR mutation and / or HER2 mutation.

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