Compounds for use as kinase inhibitors and uses thereof

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 belongs to 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] The epidermal growth factor receptor (EGFR) belongs to the receptor tyrosine kinase (RTK) family, which includes EGFR / ERBB1, HER2 / ERBB2 / NEU, HER3 / ERBB3, and HER4 / ERBB4. EGFR activates its tyrosine kinase activity through homodimerization or heterodimerization, phosphorylating its substrates and activating several downstream pathways within the cell, such as the PI3K-AKT-mTOR pathway, which is involved in cell survival, and the RAS-RAF-MEK-ERK pathway, which is involved in cell proliferation. EGFR mutations or amplifications can activate heterodimeric kinases, leading to the development of many human diseases, including malignant tumors. For example, among non-small cell lung cancer (NSCLC) patients, approximately 10% or more in the United States have EGFR mutations, while the proportion of EGFR mutations in Asia is nearly 50%. At the same time, the incidence of HER2 mutations is approximately 2-4% among NSCLC patients.

[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, currently available EGFR-TKIs include the first-generation Iressa, Tarceva, and Commana, the second-generation afatinib and dacomitinib, and the third-generation osimertinib. The remaining 10% of EGFR mutations primarily involve EGFR exons 18 and 20, and EGFR exon 20 insertion mutations account for approximately 9% of all EGFR mutations. For patients with HER2-mutated tumors, the most common HER2 mutation is the HER2 exon 20 insertion mutation.

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

[0005] To meet the clinical needs of patients with EGFR and HER2 mutations, especially those with EGFR and HER2 exon 20 insertion mutations, WO2021180238 discloses a series of compounds with excellent activity against EGFR and HER2 exon 20 insertion mutations, as well as EGFR exon 19 deletions and exon 21 L858R point mutations, and these compounds have great potential for development as drugs to treat related diseases. Among these, compounds with tripentaamine structures have been shown to be highly effective in treating various related disease mutations. Further research has shown that the configuration of the tripentaamine structure is crucial, with the transition from trans to cis improving drug efficacy in the body and providing effective treatment for different mutations. In subsequent research and development, the present invention has investigated and screened polymorphs of free base compounds to find more stable and bioavailable free base compounds.

[0006] The objective of the present invention is to provide a compound used as a kinase inhibitor, which has a cis-tripentamine structure and 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 pharmacological effect and wider applicability than the corresponding trans-structure compounds in Patent WO2021180238.

[0007] A second object of the present invention is to provide the use of the above compounds in the manufacture of a medicament for treating 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 above compounds, and by conducting polymorphism studies, a practical crystalline form with high stability and higher bioavailability is identified. [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 thereof, or a pharmaceutically 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 "pharmaceutically acceptable salt" of the drug substance is a salt that is pharmaceutically 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 hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, nitrate, phosphate, and acid phosphate. 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 4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, dodecylsulfonic 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 organic bases (e.g., ethanolamine, diethanolamine, triethanolamine, trometamol, N-methylglucamine, etc.). Typical organic acid salts are selected from 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 apparent that the pharmaceutically acceptable salts are non-toxic.

[0011] Solvates are compounds that contain a solvent, such as hydrates, dimethylsulfoxide solvates, and the like. Prodrugs are compounds that, 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 of formula 2, or a deuterated form thereof, or a pharmaceutically 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 of formula 3, or a deuterated form thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof. [ka] In Formula 3, X is selected from CH, R is -CH, -CHCH, -CHCF, and 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 the 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, as well as EGFR exon 19 deletion and exon 21 point mutations.

[0016] Preferably, the EGFR mutation and / or HER2 mutation includes one or a combination of two or more 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. Biological activity experiments have demonstrated that the above compounds have 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 and 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 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 of the crystalline form of a selected compound represented by general formula 1 was conducted. Furthermore, a polymorph screening study was conducted on the compound of Example 1 of the present invention to find a stable and reliable crystalline form, ensuring the stability of the compound's quality while allowing the drug to exert better effects in clinical treatment. The product of Example 1 was found to be a crystalline product with good crystallinity, and in an anhydrous crystalline form, named free base crystalline 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 this 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] Furthermore, the powder X-ray diffraction spectrum of 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, solvent precipitation, high-temperature cycling, and evaporative crystallization in various solvents, including, but not limited to, 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, 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 of dichloroethane and water, and all of these solvates were characterized. Competitive suspension experiments were performed on the two anhydrous crystalline forms, free base crystalline Form I and free base crystalline Form V, and it was suggested that free base crystalline Form I was a thermodynamically stable crystalline form and more suitable for further development.

[0023] The crystalline form of free base Form I was evaluated, including dry grinding, wet grinding, tableting (30 MPa), stability, and hygroscopicity studies. After 5 minutes of dry grinding, it became amorphous. After 5 minutes of wet grinding with water, there was almost no change, and after 5 minutes of wet grinding with ethanol, the crystallinity slightly decreased. Tableting tests showed a moderate decrease in crystallinity at 30 MPa. Stability studies revealed that free base Form I was relatively stable at different temperatures and humidities, with no change in liquid phase detection results. Dynamic vapor sorption (DVS) tests showed that at 80% RH, the weight gain due to water absorption was 0.66%, indicating that free base Form I is slightly hygroscopic. Therefore, free base Form I is a relatively stable anhydrous crystalline form with stable solid properties and slight hygroscopicity, making it useful for subsequent drug development. [Brief explanation of the drawings]

[0024] [Figure 1] Figure 1 shows the change in tumor volume (mm3) in the LU0387 model. [Figure 2] Figure 2 shows the change in tumor volume (mm3) in the Ba / F3 EGFR D770_N771 ins SVD model. [Figure 3] FIG. 3 is a single crystal diffraction spectrum of the single crystal of Example 1. [Figure 4] FIG. 4 is the XRPD spectrum of free base crystalline Form I. [Figure 5]FIG. 5 is an overlay of the DSC and TGA of Form I of the free base. [Figure 6] Figure 6 shows the XRPD spectrum of an amorphous sample obtained by dry grinding. [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 overlaid DSC and TGA graph of the free base crystalline Form III. [Figure 11] FIG. 11 is an XRPD spectrum of free base crystalline Form IV. [Figure 12] FIG. 12 is an overlay of DSC and TGA of 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 INVENTION

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

[0026] In the method for producing 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 reacted with each other 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 the presence of 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) Compound G is reduced 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) Hydrolysis of 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) Compound g is reduced 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 production 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 250 mL three-neck flask, under nitrogen gas protection, compound 1 (2.01 g, 8.5 mmol) was placed and dissolved in 100 mL of tetrahydrofuran. Anhydrous aluminum trichloride (2.26 g, 17 mmol) was added and stirred at 70 °C for 1 hour. Compound 2 (1.34 g, 10.2 mmol) was added dropwise. After the addition was complete, the reaction was continued at 70 °C. The reaction was monitored by sampling on a plate. After 4 hours, the reaction was almost complete. After treatment, 1.31 g of product was obtained, with a yield of 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 heated to 80 °C under N2 protection overnight. The reaction was monitored by sampling on a plate. After complete reaction and treatment, 1.50 g of product was obtained, 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 and reacted at 90°C overnight. The mixture was sampled and monitored on a plate. After the reaction was complete, the mixture was treated 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 reaction mixture was sampled and monitored on a plate. After the raw materials had completely reacted, the mixture was treated to obtain 7.0 g of the product.

[0039] Synthesis of Example 1: Compound 8 (7.00 g, 12.6 mmol) and triethylamine (3.82 g, 37.8 mmol) were dissolved in dichloromethane and cooled to 0°C under nitrogen gas protection. A solution of acryloyl chloride (1.72 g, 18.9 mmol) in dichloromethane was added dropwise and allowed to react for 1 hour while maintaining the temperature at 0°C. A sample was placed on a plate for monitoring. After the raw materials were completely reacted, the mixture was processed. Specifically, aqueous sodium bicarbonate solution and dichloromethane were added, and the mixture was stirred to separate the layers. The aqueous phase was further extracted once with dichloromethane. The organic phases were combined, dried, rotary evaporated, and passed through a column. The resulting 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. This was filtered, and the cake was directly added to methanol, mixed, and dried, yielding 3.5 g of product. [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] Referring to Example 1, three deuterated compounds in Examples 2, 3 and 4 were synthesized, and the details are 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: Compound 1 (54.6 g, 303 mmol) and acetohydrazide (26.64 g, 395 mmol) were placed in a 2000 mL single-neck flask, and 1000 mL of 1N aqueous sodium hydroxide solution was added. The mixture was allowed to react at 80°C for 4 hours, resulting in the precipitation of a large amount of solid. The temperature was lowered, and 80 mL of concentrated hydrochloric acid was added. The mixture was stirred at a low temperature of approximately 0°C for 30 minutes, then suction filtered. The cake was stirred once with water and vacuum dried overnight at 55°C to obtain 32 g 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: Compound 2 (50.0 g, 258 mmol) was placed in a 2000 mL single-neck flask, and 1500 mL of toluene, phosphoryl chloride (237 g, 1550 mmol), and N,N-diisopropylethylamine (133 g, 1031 mmol) were added at room temperature. A white mist was generated. 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 30 g of the product. 1 H NMR (400 MHz, chloroform-d) δ 9.19 (s, 1H), 2.71 (s, 3H).

[0045] Synthesis of Compound 5: Compound 3 (30.0 g, 130 mmol) was placed in a 2000 mL single-neck flask and dissolved completely in 1500 mL of 1,2-dichloroethane. Anhydrous aluminum trichloride (29.3 g, 220 mmol) was added and stirred at room temperature for 30 minutes. The temperature was lowered to 0°C, and compound 4 (22.1 g, 169 mmol) was added dropwise. The mixture was stirred at low temperature for 30 minutes, then heated to 60°C for reaction. The reaction mixture was sampled and monitored on a plate. After 4 hours of reaction, the temperature was lowered and the mixture was processed to obtain 20.0 g 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 reaction was monitored by sampling on a plate, and after completion of the reaction, the product was treated to obtain 0.65 g.

[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 and monitored on a plate. After the raw materials had completely reacted, the mixture 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 added to 50 mL of methanol and reacted at room temperature for 4 hours in a hydrogen gas atmosphere. After the reaction was complete, the mixture was sampled and monitored on a plate, and 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 cooled to 0°C under 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 reaction mixture was sampled and monitored on a plate. After the reaction of the raw materials was complete, the mixture was processed to obtain 200 mg of the product. [M+H] + :606.8.

[0050] Referring 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 and monitored on a plate. After the raw materials had completely reacted, the mixture was processed 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 6:3:1 volumetric ratio mixed solvent of tetrahydrofuran, methanol, and water and reacted at 40°C for 16 hours. After the raw materials were completely reacted, the mixture was sampled and monitored. After the reaction was complete, 6.02 g of the product was obtained.

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

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

[0057] Synthesis of Compound 7: Compound 6 (1.0 g, 1.68 mmol) and triethylamine (679 mg, 6.72 mmol) were added to 20 mL of dichloromethane and stirred for 15 minutes. Trifluoroacetic anhydride (1.59 g, 7.58 mmol) was then added in portions and the reaction was continued at 40°C for 8 hours until the reaction was complete. After further treatment, 521 mg of the 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 the mixture was 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 had completely reacted, the mixture was treated to obtain 412 g of the product.

[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 cooled to 0°C under a nitrogen gas atmosphere. A dichloromethane solution of acrylic chloride (93 mg, 1.02 mmol) was then added dropwise and the reaction was continued for 2 hours while maintaining the temperature at 0°C. The reaction mixture was sampled and monitored on a plate. After the reaction was complete, the mixture was processed to obtain 122 mg of 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 were synthesized in Examples 10, 11, and 12, which are specifically 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: Compound 1 (10.00 g, 62.88 mmol) was placed in a 500 mL single-neck flask and completely dissolved in 100 mL of tetrahydrofuran. Cesium carbonate (20.50 g, 62.88 mmol) and trifluoroethanol (6.29 g, 62.87 mmol) were added, and after the addition was complete, the flask was protected with nitrogen gas. The reaction was monitored by sampling on a plate and allowed to proceed at 23 °C for 6 hours, until the reaction was nearly complete. After treatment, 14.35 g of product was obtained, 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 (14.00 g, 58.57 mmol) was dissolved in 60 mL of ethanol, and 15 mL of water, ammonium chloride (9.60 g, 179.44 mmol), and reduced iron powder (20.00 g, 357.14 mmol) were added. After the addition was complete, the mixture was heated to 80 °C and reacted overnight. The reaction was monitored by sampling on a plate. After the reaction was complete, 10.50 g of product was obtained, 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: Compound 3 (10.00 g, 47.83 mmol) was placed in a 250 mL three-neck flask. 50 mL of concentrated sulfuric acid was added while stirring. The temperature was lowered to 0 °C, and solid potassium nitrate (6.10 g, 60.34 mmol) was added in portions. After the addition was complete, the flask was protected with nitrogen gas. After the reaction was allowed to proceed at room temperature for 4 hours, the mixture was sampled on a plate and treated to obtain 8.26 g of the product, 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. The mixture was stirred overnight at 80 °C. The next day, a sample was placed on a plate for monitoring, and the reaction mixture was almost completely completed. The temperature was lowered, and a solid precipitated from the reaction mixture. The solid was filtered, and the cake was spun dry. Methanol was added and mixed to obtain 1.67 g of the 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. Samples were placed on a plate for monitoring, and after the raw materials had completely reacted, the mixture was processed to obtain 1.16 g of the 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 of methanol and 20 mL of ethyl acetate, and the mixture was reacted at room temperature for 3 hours in a hydrogen gas atmosphere. After the reaction was complete, the mixture was sampled and monitored on a plate, and 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 cooled 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 reaction was continued for 2 hours while maintaining the temperature at 0°C. The reaction mixture was sampled and monitored on a plate. After the reaction was complete, the mixture was processed to obtain 170 mg of 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: Compound 2 (112 mg, 1.24 mmol), 20 mL of dichloromethane, and oxalyl chloride (142 mg, 1.12 mmol) were placed in a 50 mL single-neck flask and stirred under nitrogen gas for 2 hours. The mixture was cooled to 0°C, and a dichloromethane solution of compound 1 (438 mg, 78.92 mmol) and triethylamine (152 mg, 1.50 mmol) were added dropwise. The mixture was allowed to react for 1 hour while maintaining the temperature at 0°C. Samples were collected on a plate for monitoring. After the raw materials had completely reacted, the mixture was processed, and 160 mg 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: Compound 1 (313 mg, 1 mmol) was placed in a 100 mL single-neck flask, followed by 5 mL of acetonitrile and then the crude product of Compound 2 (441 mg). After the addition was complete, the flask was protected with nitrogen gas. The reaction was monitored by sampling onto a plate, and the mixture was allowed to react overnight at 80°C until the reaction was nearly complete. After further treatment, 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 complete, the mixture was heated to 60°C and reacted overnight. The reaction was monitored by sampling on a plate. After the reaction was complete, 0.89 g of the product was obtained after treatment. 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 under nitrogen gas protection. The reaction was monitored by sampling on a plate, and after completion of the reaction, the mixture 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 of methanol and 2 mL of ethyl acetate, and the mixture was reacted at room temperature for 2 hours in a hydrogen gas atmosphere. After the raw materials were completely reacted, the mixture was sampled and monitored on a plate, and 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 cooled to 0°C under a nitrogen gas atmosphere. A dichloromethane solution of acryloyl chloride (70 mg, 0.77 mmol) was then added dropwise and the reaction was continued for 2 hours while maintaining the temperature at 0°C. The reaction mixture was sampled and monitored on a plate. After the reaction was complete, the mixture was processed to obtain 118 mg of 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: Compound 1 (153 mg, 0.38 mmol) was placed in a 100 mL three-neck flask, followed by 5 mL of tetrahydrofuran and 4-dimethylaminopyridine (17.8 mg). After the addition was complete, di-t-butyl dicarboxylate (1.06 g, 4.86 mmol) was added dropwise under nitrogen gas protection. The reaction mixture was sampled and monitored on a plate, and the reaction was continued at 80°C for 2 hours until the reaction was nearly complete. After further processing, 104 mg of the 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 a 5.4 mol / L solution of sodium methoxide in methanol was added. After the addition was complete, the reaction was allowed to proceed for 30 minutes. The reaction was monitored by sampling on a plate. After the reaction was complete, 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 of methanol and 1 mL of ethyl acetate, and the mixture was reacted at room temperature for 2 hours in a hydrogen gas atmosphere. After the reaction was complete, the mixture was sampled and monitored on a plate, and 30 mg of the product was obtained.

[0086] Synthesis of compound 5: Compound 4 (30 mg, 0.069 mmol) and triethylamine (13 mg, 0.14 mmol) were dissolved in dichloromethane and cooled 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 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 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. The mixture was heated to 100°C in a nitrogen gas atmosphere and reacted for 4 hours. The mixture was sampled and monitored on a plate. After the raw materials had completely reacted, the mixture 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 selected in the experiment, 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, and Ba / F3-HER2-A775-G776 ins YVMA, were provided by Hefei Zhongke Pu Ruisheng Biopharmaceutical Technology Co., Ltd. and were validated before use in this study. A 20x stock solution of test compound was prepared for use, diluted in 3-fold gradient dilutions at nine concentrations starting from 1 μM.

[0089] Logarithmic phase cell suspensions were seeded into 96-well white cell culture plates, each with a volume of 95 μL (2000 cells / well). 5 μL of 20x test compound was added to each culture plate containing the 95 μL cell suspension, referring to the plate seeding diagram, and mixed evenly. The mixture was incubated in a 37°C, 5% CO2 incubator for 72 hours. The growth inhibitory activity of the compounds was measured using the CellTiter-Glo method. The corresponding fluorescence value (RLU) for each well was obtained from the SpectraMax Paradigm readings. The inhibition rate data was calculated using the following formula: Inhibition Rate (Inh%) = 100 - (RLU compound - RLU blank) / (RLU control - RLU blank) * 100%. The cell viability corresponding to different concentrations of the compound was calculated in Excel, and the cell viability curve was plotted using GraphPad Prism software to determine 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 magnitude of the value, where A≦30nM, 30nM<B≦100nM、C> It is 100nM. Table 6. Test results of the cytostatic activity of different compounds on EGFR exon 20 insertion mutations [Table 6]

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

[0093] Table 6 shows that the compounds in the examples all have good inhibitory activity against EGFR exon 20 insertion mutations, superior to DZD9008 and comparable to the activity of TAK-788. Table 7 shows that the cellular activity of Example 5 against HER2 exon 20 insertion mutations is comparable to TAK-788 and better than DZD9008.

[0094] 1.2 Testing the inhibitory activity of compounds on 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 10x stock solution of test compound was prepared for use, diluted from 10 μM in 4-fold gradient dilutions at nine concentrations.

[0095] The logarithmic phase cell suspension was taken and inoculated into a 96-well culture plate, with each well containing 90 μL (2000 cells / well). 10 μL of 10x test compound was taken and added to each culture plate, referring to the plate inoculation diagram, and mixed evenly. The cells were incubated in an incubator at 37°C with 5% CO2. The growth inhibitory activity of the compounds was measured using the CellTiter-Glo method. The cell growth inhibition rate data was calculated by plotting a cell activity curve using 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 experiments of compounds 2.1 Pharmacodynamic studies on the LU0387 PDX lung cancer model The lung cancer LU0387PDX model is a commonly used xenograft tumor model in coronary artery surgery. The experimental conditions for this model are as follows: After euthanizing the tumor-bearing mice, the tumor masses were removed under aseptic conditions, washed, and blood stains, connective tissue, and necrotic areas were removed. The tumors were then divided into 2×2×2mm x ... 3 The tumor was cut into small pieces and inoculated subcutaneously into the right dorsal region of the sterilized experimental mice using an inoculation needle. The tumor growth was monitored periodically until the tumor reached an average volume of 100-200 mm. 3 Once the tumors had grown to 100 mg / kg, the mice were randomly assigned to groups based on tumor size and body weight and administered the treatment. Before the start of treatment, all animals were weighed and tumor diameters were measured with a vernier caliper. To ensure that tumor volumes were comparable across groups, the mice were randomly assigned to groups based on tumor volume. Each group contained three mice, and the mice were orally administered the treatment once daily. After administration, the inoculation site was observed. Tumor volume and mouse body weight 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 Figure 1. 1 shows that in the LU0387 model, the efficacy of Example 1 was better than that of TAK-788 and DZD9008 at the same dose. The efficacy of Example 5 was similar to that of TAK-788. With increasing doses of Example 1, the efficacy was also significantly 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 Ba / F3 EGFR D770_N771 ins SVD cell line 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 required numbers. 0.2 ml (1 x 106 cells) of cells were inoculated subcutaneously into the right dorsal region of 6-8 week-old female nude mice weighing 18-22 g. The average tumor volume was approximately 150-200 mm. 3 Once the tumor volume reached 1000 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 vernier caliper. To ensure that tumor volumes were comparable across groups, mice were randomly assigned to groups based on tumor volume. Each group contained three mice, and the mice were orally administered the drug once daily. After administration, the inoculation site was observed. 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 Figure 2. Figure 2 shows that in the Ba / F3-EGFR-D770_N771 ins SVD cell transplant model, at the same dosage, the internal efficacy of both Example 1 and Example 5 was far superior to that of the compound having a trans-tripentamine structure in Patent WO2021180238, with the tumor inhibition rate 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 mutations, and also have good inhibitory effects on cell proliferation in the cell lines NCI-H1975 and PC9. For one or a combination of two or more of EGFR exon 20 insertion mutations, HER2 exon 20 insertion mutations, EGFR exon 19 deletions, EGFR exon 20 point mutations, and EGFR exon 21 point mutations, these drugs have good therapeutic effects on related diseases.

[0108] 3. Structural confirmation of Example 1 and study of the free base crystalline form [ka]

[0109] 1. Structure confirmation 214 mg of the compound of Example 1 was taken, and 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 sealed sample bottle, and after piercing, it was slowly evaporated in a draft chamber to obtain granular crystals (free base crystalline form I). The structure was confirmed by single crystal diffraction, and the results are shown in Figure 3, confirming the configuration of the compound.

[0110] 2. Study of the Free Base Crystalline Form of Example 1 Polymorph screening was performed on the compound obtained in Example 1 to identify potential crystalline forms suitable for further development. Using the synthesized Example 1 as the starting material, the compound was found to be crystalline with good crystallinity and in an anhydrous crystalline form, which was named free base crystalline form I. An amorphous free base compound was then prepared by dry grinding. Using these two starting materials, a crystalline form screening experiment was conducted.

[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) revealed that the raw material consisted of irregularly shaped crystals with good crystallinity. Figure 4 shows the XRPD spectrum of the free base crystalline form I, and Figure 5 shows the overlaid DSC and TGA graphs of the crystalline form. The differential scanning calorimetry (DSC) curve showed a single endothermic peak at 230°C, which corresponds to the melting peak. The thermogravimetric analysis (TGA) curve showed no significant weight loss before decomposition, indicating that it was 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 roughly measured by visual inspection in 18 different solvents, and 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 tested.

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

[0115] 2.1.3 Preparation and Characterization of Amorphous Raw Materials 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, and the XRPD detection result is shown in Figure 6.

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

[0119] 2.2.3 Differential scanning calorimetry (DSC) The DSC curve was obtained using a TA DSC 250. The measurement method for the DSC 250 was to place an appropriate amount of sample in an aluminum crucible (precisely weighed and drilled), and then heat the crucible from 25°C to a final temperature of 300°C at a rate of 10°C / min, followed by blowing with nitrogen gas at a flow rate of 50 mL / min.

[0120] 2.2.4 Thermogravimetric analysis (TGA) TGA data were obtained using a TA instrument, model TGA 550. An appropriate amount of sample was placed in an 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 Free Base Polymorphic Study of the Compound of Example 1 They were mainly prepared by methods such as suspension crystallization, anti-solvent precipitation, high-temperature circulation and evaporation crystallization. Based on the results of visually determining the solubility of the compound in Example 1, suspension crystallization was carried out using free base crystalline Form I and amorphous crystalline Form as starting materials at 25°C and 50°C, respectively, in a single solvent (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) or a mixture of solvents. A total of two crystalline forms were obtained: free base crystalline Form I and free base crystalline Form II.

[0122] Based on the results of visually determining the solubility of the compound in Example 1, anti-solvent precipitation was carried out by selecting dichloromethane and dichloroethane as good solvents, and adding different anti-solvents to each of these solvents under stirring at room temperature (up to 25°C). Two types of crystalline forms were obtained: free base crystalline form I and free base crystalline form III.

[0123] The high-temperature cycling method involves adding a solvent (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) to the compound, and then stirring under temperature cycling conditions of 50°C to 5°C to produce the crystalline form. A total of two crystalline forms were obtained: free base crystalline form I and free base crystalline form II.

[0124] Evaporative crystallization was performed by adding a solvent to the compound, filtering the resulting solution, and dripping the filtrate into a sample bottle. After puncturing the sealing membrane covering the opening, the solution was slowly evaporated in a fume hood. Two crystalline forms were obtained: free base crystalline form I and free base crystalline form IV. After heating free base crystalline form IV to 150 °C to remove the solvent, a new crystalline form was prepared, designated free base crystalline form V.

[0125] 2.3.1 Preparation and Characterization of Free Base Form I Free base crystalline form I is obtained in most solvents, and the compound obtained in Example 1 is 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 Form II of the free base was obtained in a solvent system containing a portion of ethanol. Form II of the free base was obtained by suspension crystallization of the amorphous compound in a single solvent, ethanol. Its characterization results are shown in Figures 7 and 8. The DSC curve of Form II of the free base exhibits two endothermic peaks at 124°C and 230°C, and the TGA curve exhibits a weight loss of 4.48% between 65°C and 150°C. Form II of the free base was obtained in an ethanol solvent system. From the above, it is inferred that Form II of the free base is a solvate of ethanol. The endothermic peak at 124°C in the DSC curve corresponds to 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 Form III of the free base was obtained in a solvent system containing a small amount of isopropanol. When the crystalline form was prepared by anti-solvent precipitation, Form III of the free base was observed in dichloromethane / isopropanol or dichloroethane / isopropanol systems. The characterization results are shown in Figures 9 and 10. The DSC curve of Form III of the free base exhibited two endothermic peaks at 121°C and 230°C, the TGA curve showed a weight loss of 9.57% between 75°C and 145°C, and after heating to 150°C and desolvation, Form III of the free base was transformed to Form I of the free base by XRPD. Form III of the free base was obtained in a solvent system containing isopropanol. From the above, it is inferred that Form III of the free base 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 Form IV of the free base was obtained in a solvent system containing a portion of dichloroethane. 92.28 mg of the starting material was placed in a sample vial, and 5.0 mL of dichloroethane was added to dissolve the material. The vial was then covered with a sealing film and pierced, after which the material was slowly evaporated in a fume hood to produce Form IV by evaporative crystallization. The characterization results are shown in Figures 11 and 12. The DSC curve of Form IV of the free base showed two endothermic peaks at 108°C and 231°C, and the TGA curve showed a weight loss of 10.41% from 25 to 150°C.1 The H NMR results indicated that only 6.14% of the dichloroethane solvent remained, suggesting that the sample contained 4.27% water. From the above, it is speculated 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 free base crystalline form IV to 150°C to remove the solvent, it was characterized by XRPD, which showed that it had formed 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 had an endothermic peak at 231°C, and the TGA curve showed 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 free base Form I During this screening process, only two anhydrous crystalline forms, free base crystalline Form I and free base crystalline Form V, were discovered. Saturated solutions were prepared using two solvents, acetone and acetonitrile, and competitive suspension of the two was performed at 25°C and 50°C. As a result, free base crystalline Form I was inferred to be the thermodynamically stable crystalline form of the compound. Furthermore, free base crystalline Form V was not directly obtained in the screening experiment, but was obtained only by desolvation of free base crystalline Form IV. Therefore, evaluation of the crystalline form was performed only on free base crystalline Form I, including studies on dry grinding, wet grinding, tableting (30 MPa), stability, and hygroscopicity.

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

[0136] In the stability experiment, free base crystalline Form I was stored open at 80°C for 3 days, 60°C for 7 days, 25°C / 60%RH for 7 days, 40°C / 75%RH for 7 days, and 25°C / 90±5%RH for 7 days. Samples were taken and subjected to XRPD detection. The results showed that there was no obvious change in free base crystalline Form I, and the liquid phase detection results of the drug also showed no change.

[0137] Dynamic vapor sorption (DVS) testing of Form I of the free base showed that the weight gain upon absorption of water was 0.66% at 80% RH, indicating that Form I of the free base 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. 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 with stable solid properties and slight hygroscopicity, which is useful for subsequent drug development.

Claims

1. A compound used as a kinase inhibitor, characterized in that the compound is a compound represented by formula 1, or a deuterated product thereof, or a pharmaceutically acceptable salt or solvate thereof. 【Chemical 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 product thereof, or a pharmaceutically acceptable salt or solvate thereof. 【Chemistry 3】

3. In Formula 2, X is selected from CH and N, and 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 product thereof, or a pharmaceutically acceptable salt or solvate thereof. 【Chemistry 4】

5. In Formula 3, X is selected from CH and N, and 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 group consisting of:

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

7. The compound used as a kinase inhibitor according to claim 1, characterized in that 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 of formula A, 【Chemistry 6】 The compound is in crystalline form 8. The compound for use as a kinase inhibitor according to claim 7, wherein the crystalline form is free base crystalline form I, and the powder X-ray diffraction spectrum of said crystalline form shows characteristic diffraction peaks at 2θ angles 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 angles 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°, and 15.63°±0.2°. The compound used as a kinase inhibitor according to 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 treating 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 includes one or a combination of two or more of an EGFR exon 20 insertion mutation, a HER2 exon 20 insertion mutation, an EGFR exon 19 deletion, a point mutation in EGFR exon 21, and a point mutation in EGFR exon 20.

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, characterized in that the disease is a cancer caused by the EGFR mutation and / or HER2 mutation.

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