2,5-disubstituted thiazole compounds containing a pyrimidine ring, and preparation method and application thereof

Abstract

The application discloses a 2,5-disubstituted thiazole compound containing a pyrimidine ring and a preparation method and application thereof, and belongs to the technical field of medicines. The application provides a 2,5-disubstituted thiazole compound containing a pyrimidine ring or a pharmaceutically acceptable salt thereof, a preparation method of the 2,5-disubstituted thiazole compound containing a pyrimidine ring or the pharmaceutically acceptable salt thereof, and application of the compound or the pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound in preparation of a medicine for treating a disease related to abnormal expression of anaplastic lymphoma kinase.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to novel pyrimidine-ring-containing 2,5-disubstituted thiadiazole compounds, their preparation methods, and the use of said compounds or pharmaceutically acceptable salts thereof or pharmaceutical compositions containing said compounds in the preparation of medicaments for treating diseases associated with abnormal expression of anaplastic lymphoma kinase. Background Technology

[0002] Anaplastic lymphoma kinase (ALK) is a highly conserved transmembrane receptor tyrosine kinase belonging structurally to the insulin receptor (IR) kinase superfamily. Structurally, ALK consists of an N-terminal extracellular domain, a hydrophobic one-way transmembrane region, and an intracellular kinase domain. When the ALKAL protein (the endogenous ligand of ALK) binds to its extracellular domain, ALK is activated, leading to dimerization and autophosphorylation, thereby disrupting cell proliferation and survival. In 1994, Morris et al. discovered NPM1-ALK gene rearrangement mutations in patients with anaplastic large cell lymphoma. However, in 2007, Soda et al. first discovered EML4-ALK gene rearrangement mutations in patients with non-small cell lung cancer (NSCLC). ALK mutations lead to constitutive abnormal activation of ALK kinase and related downstream signaling pathways such as RAS-MAPK, PI3K-AKT, and JAK-STAT.

[0003] Since the first report of EML4-ALK gene rearrangement mutations in NSCLC patients, approximately 3%-7% of NSCLC patients have ALK rearrangements. Although the proportion of ALK rearrangements is relatively small, the large number of NSCLC cases results in about 40,000 new cases of this type worldwide each year. Currently, ALK inhibitors have maintained a high level of development and research interest in targeted therapy for NSCLC. As the pathogenesis and drug resistance mechanisms continue to be elucidated, new ALK inhibitors are constantly emerging, with small molecule ALK inhibitors undergoing four generations of drug development.

[0004] Crizotinib, developed by Pfizer, is a first-generation ALK inhibitor approved by the FDA in 2011 for the first-line treatment of ALK-rearranged NSCLC. In phase I / II clinical trials, the drug demonstrated significant efficacy in patients with advanced ALK-rearranged NSCLC (ORR approximately 60%). Subsequent randomized phase III trials showed that crizotinib was superior to chemotherapy in treating ALK-rearranged NSCLC. However, similar to early EGFR TKIs, acquired resistance to crizotinib is common, typically developing within one year of treatment initiation (median progression-free survival 7.7–10.9 months). To address the resistance issue of first-generation ALK inhibitors, more effective second-generation ALK inhibitors have been developed, including ceritinib, alectinib, and brigatinib. However, like crizotinib, patients eventually develop resistance to second-generation ALK inhibitors. Mutations mainly occur in the solvation front region and the C-terminal region of the α-C helix, such as G1202R and F1174L mutations. Among them, the G1202R solvation region mutation, where glycine residues are replaced by larger arginine residues, is one of the most common gene mutations in second-generation ALK inhibitors, accounting for 35%-60%. Lorlatinib is a third-generation ALK inhibitor with a broad-spectrum anti-ALK kinase domain and was approved in March 2021 for patients with advanced ALK rearrangement NSCLC. Although lorlatinib has shown good clinical efficacy, acquired resistance has inevitably emerged. Studies have shown that most target ALK mutations that lead to lorlatinib resistance are complex ALK mutations (such as C1156Y / L1198F, G1202R / L1196M, I1171N / D1203N, etc.). TPX-0131 and NVL-655 are representative fourth-generation ALK inhibitors, currently in Phase I clinical trials. Clinical data show that they are highly sensitive to single or multiple resistance mutations of approved ALK inhibitors (such as G1202R+L1196M, G1202R+G1269A, G1202R+L1198F, etc.).

[0005] While fourth-generation ALK inhibitors have overcome some ALK compound mutations, various resistance mutations continue to emerge against different inhibitors. Currently, existing ALK inhibitor treatment strategies are insufficient to meet clinical needs. Given the susceptibility to ALK active site mutations, there is still an urgent need to develop novel ALK inhibitors that can overcome resistance mutations for the treatment of non-small cell lung cancer. Summary of the Invention

[0006] The primary objective of this invention is to provide a novel pyrimidine-ring-containing 2,5-disubstituted thiadiazole compound as shown in general formula I, and a method for preparing the same. Additionally, the invention provides the use of said compound, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing said compound in the preparation of medicaments for treating conditions associated with anaplastic lymphoma kinase mutations.

[0007]

[0008] in:

[0009] X is selected from C and N;

[0010] R1 is selected from H, (C1-C6)alkyl, (C1-C6)alkoxy, halogen, halo(C1-C6)alkyl, nitro; R1 can be one or more;

[0011] A is selected from 5-6 aryl or heteroaryl, wherein the heteroaryl contains 1-3 heteroatoms optionally selected from N, O and S;

[0012] R2 is selected from H, (C1-C6)alkyl, (C1-C6)alkoxy, halogen, (C1-C6)alkylsulfonyl, (C1-C6)alkylamide; or R2 together with the carbon atom on A to form a 4-6 membered cycloalkyl or aryl group;

[0013] R3 is selected from H, (C1-C6)alkyl, (C1-C6)alkoxy, halogen, cyano, nitro, hydroxyl, amino,

[0014]

[0015] R4 and R5 may be the same or different, and are independently selected from H, (C1-C6)alkyl, (C1-C6)alkoxy, hydroxyl, or hydroxyl-substituted (C1-C6)alkyl; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group, wherein the heterocyclic group optionally contains 0-1 heteroatoms selected from N, O, and S in addition to the nitrogen atom attached to R4 and R5, and the heterocyclic group is optionally substituted by 0-1 R6 atoms;

[0016] R6 is selected from H, halogen, cyano, nitro, hydroxyl, carboxyl, amino, (C1-C6)alkyl, hydroxyl or halogen or amino or cyano or carboxyl-substituted (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)alkylamino, di(C1-C6)alkylamino, (C1-C6)alkylcarbonyl, (C1-C6)alkylsulfonyl, (C1-C6)alkylsulfonylamino, (C1-C6)alkylamide.

[0017] Furthermore, in the general formula I:

[0018] R1 is selected from H, (C1-C6)alkyl, (C1-C6)alkoxy, and halogen; R1 can be one or more;

[0019] A is selected from 5-6 aryl or heteroaryl, wherein the heteroaryl contains 0-1 heteroatom selected from N, O and S;

[0020] R2 is selected from H, (C1-C6)alkylsulfonyl, (C1-C6)alkylamide; or R2 together with the carbon atom on A to form a 4-6 membered cycloalkyl group;

[0021] R3 is selected from H, (C1-C6)alkyl, amino,

[0022] R4 and R5 may be the same or different, and are independently selected from H, (C1-C6)alkyl, hydroxyl, or hydroxyl-substituted (C1-C6)alkyl; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group, wherein the heterocyclic group, in addition to the nitrogen atom attached to R4 and R5, optionally contains 0-1 heteroatoms selected from N, O, and S, and the heterocyclic group is optionally substituted by 0-1 R6;

[0023] R6 is selected from H, hydroxyl, amino, (C1-C6)alkyl, hydroxyl or halogen or amino or cyano or carboxyl-substituted (C1-C6)alkyl, (C1-C6)alkylamino, bis(C1-C6)alkylamino.

[0024] Furthermore, in the general formula I:

[0025] R1 is selected from H, (C1-C6)alkoxy, or halogen; there can be one or more R1s.

[0026] A is selected from 5-6 aryl groups;

[0027] R2 is selected from H, (C1-C3)alkylsulfonyl, (C1-C3)alkylamide; or R2 together with the carbon atom on A to form a 4-6 membered cycloalkyl group;

[0028] R3 is selected from H, methyl, amino,

[0029] R4 and R5 may be the same or different, and are independently selected from H, (C1-C6)alkyl, or hydroxyl-substituted (C1-C6)alkyl; or R4 and R5 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group, wherein the heterocyclic group, in addition to the nitrogen atom attached to R4 and R5, optionally contains 0-1 heteroatoms selected from N, O, and S, and the heterocyclic group is optionally substituted by 0-1 R6.

[0030] R6 is selected from H, hydroxyl, (C1-C6)alkyl, hydroxy-substituted (C1-C6)alkyl, and di(C1-C6)alkylamino.

[0031] Furthermore, in the general formula I:

[0032] R1 is selected from H, methoxy, and Cl; R1 can be one or more;

[0033] A is a phenyl group;

[0034] R2 is selected from H, isopropylsulfonyl, methylamide; or R2 together with the carbon atom on A forms cyclopentane;

[0035] R3 is selected from H, methyl, amino, and the following structures:

[0036]

[0037] Furthermore, the pyrimidine-ring-containing 2,5-disubstituted thiadiazole compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing said compound, wherein the pyrimidine-ring-containing 2,5-disubstituted thiadiazole compound is any one of compounds I-1 to I-32 listed below:

[0038]

[0039]

[0040]

[0041] The pharmaceutically acceptable salts described in this invention include addition salts formed by inorganic acids and organic acids with the 2,5-disubstituted thiadiazole compounds containing a pyrimidine ring, wherein the inorganic acids and organic acids include: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, thearubigin, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, and benzoic acid.

[0042] In this invention, "halogen" refers to fluorine, chlorine, bromine, or iodide; "alkyl" refers to a straight-chain or branched alkyl group; and "heterocyclic group" refers to a cyclic system containing one or more monocyclic or polycyclic heteroatoms selected from N, O, and S heteroatoms.

[0043] This invention uses 2,5-disubstituted thiadiazole compounds containing a pyrimidine ring, or pharmaceutically acceptable salts thereof, as active ingredients, mixed with pharmaceutically acceptable excipients to prepare compositions, and then formulated into clinically acceptable dosage forms. The excipients refer to diluents, adjuvants, or carriers that can be used in the pharmaceutical field. The dosage forms include commonly used clinical formulations such as injections, tablets, capsules, aerosols, suppositories, films, pellets, topical liniments, and ointments.

[0044] The 2,5-disubstituted thiadiazole compounds containing a pyrimidine ring described in this invention have a significant inhibitory effect on ALK kinase, especially on ALK, which is closely related to the development of drug resistance. G1202 With ALK L1196M Mutations have a strong inhibitory effect.

[0045] The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring or a pharmaceutical composition containing the compound is specifically used in the preparation of a drug that inhibits ALK mutants, wherein the ALK mutant is one or more of G1202R, L1196M, G1269A, L1198F, and F1174L.

[0046] The present invention relates to the use of 2,5-disubstituted thiadiazole compounds containing a pyrimidine ring or pharmaceutical compositions comprising such compounds in the preparation of medicaments for treating cancer, wherein the cancer is preferably non-small cell lung cancer.

[0047] The following synthetic route describes the preparation of some of the pyrimidine-containing 2,5-disubstituted thiadiazole compounds of the present invention.

[0048] The synthetic route of the present invention includes the following steps:

[0049] Synthesis Route 1:

[0050]

[0051] In the process of preparing compounds I-10 to I-32 by reacting intermediate compound E with aliphatic amines, the aliphatic amines used are selected from the following compounds:

[0052]

[0053] The preparation method of intermediate A is as follows: synthetic route;

[0054] Synthesis Route 2:

[0055]

[0056] The preparation method of intermediate B is as follows: synthetic route;

[0057] Synthesis Route 3:

[0058]

[0059] In the above synthetic routes 1-3, R1, R2, R3, A, and X are the corresponding groups at the corresponding positions of compounds I-1 to I-32.

[0060] The beneficial effects of this invention are:

[0061] The compounds of this invention have novel chemical structures and exhibit high inhibitory activity against ALK mutant kinases in in vitro studies, making them suitable for the treatment and prevention of various diseases, including cancer. Detailed Implementation

[0062] The following examples provide methods for preparing some of the compounds represented by general formula I. It should be understood that the following methods, as well as other methods known to those skilled in the art, are applicable to the preparation of all compounds described herein. The examples are intended to illustrate, but not limit, the scope of the invention.

[0063] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents and instruments used, unless otherwise specified, are all commercially available conventional reagent products.

[0064] Example 1: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-1) was prepared using synthetic routes 1-3.

[0065]

[0066] Step 1: 2,5-Dichloro-N-(2-(isopropylsulfonyl)phenyl)pyrimidin-4-amine (Intermediate A1)

[0067] 2-(isopropylsulfonyl)aniline (1.09 g, 5.48 mmol) and sodium hydride (1.05 g, 43.8 mmol) were added to 15 mL of dry N,N-dimethylformamide, and the mixture was stirred at 0 °C for 0.5 h under a nitrogen atmosphere. Then, 2,4,5-trichloropyrimidine (2.00 g, 11.0 mmol) dissolved in dry N,N-dimethylformamide was slowly added, and the mixture was reacted at room temperature for 12 h. After the reaction was complete, the reaction solution was added to cold water, and a solid precipitated. The solid was filtered, and the filter cake was dried to give 3.30 g of a yellowish-brown solid, with a yield of 86.9%.

[0068] Step 2: 5-((4-amino-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl) tert-butyl carbamate (intermediate B1)

[0069] Step 2.1: 5-((3-methoxy-4-nitrophenyl)thio)-1,3,4-thiadiazole-2-amine (intermediate d1)

[0070] 5-Fluoro-2-nitrosoanisole (1.00 g, 5.84 mmol), 2-amino-5-mercapto-1,3,4-thiadiazole (1.56 g, 11.7 mmol), and potassium carbonate (1.63 g, 11.7 mmol) were added to N,N-dimethylformamide (20 mL), and the mixture was stirred at 50 °C for 4 hours. After the reaction was complete, the reaction solution was added to cold water, and a solid precipitated. The solid was filtered, the filter cake was washed with cold water, and dried to obtain 1.53 g of a yellow solid, with a yield of 92.0%.

[0071] Step 2.2: 5-((3-methoxy-4-nitrophenyl)thio)-1,3,4-thiadiazol-2-yl)tert-butyl carbamate (intermediate e1)

[0072] Intermediate d1 (1.00 g, 3.52 mmol) and di-tert-butyl dicarbonate (0.92 g, 4.22 mmol) were added to tetrahydrofuran (20 mL) and stirred at room temperature for 4 hours. After the reaction was complete, the reaction solution was poured into water (30 mL), and the solution was adjusted to alkalinity with saturated potassium carbonate solution. The mixture was extracted with dichloromethane (30 mL × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated to give 0.62 g of a yellow oily substance, with a yield of 83.5%.

[0073] Step 2.3: 5-((4-amino-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl) tert-butyl carbamate (intermediate B1)

[0074] Intermediate e1 (1.00 g, 2.60 mmol) and palladium on carbon (0.10 g) were added to a 1:10 mixture of dichloromethane and methanol (20 mL). The mixture was stirred at room temperature for 12 h in a hydrogen atmosphere. After the reaction was complete, the reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, and the filtrate was evaporated to dryness to give 0.78 g of a dark brown solid, with a yield of 84.6%.

[0075] Step 3: (5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)tert-butyl carbamate (intermediate C1)

[0076] A1 (0.59 g, 1.70 mmol), B1 (0.50 g, 1.41 mmol), lithium tert-butoxide (0.17 g, 2.12 mmol), and tetrakis(triphenylphosphine)palladium (0.10 g) were added to 1,4-dioxane (15 mL). The reaction mixture was heated to 100 °C and stirred for 10 hours under nitrogen atmosphere. After the reaction was complete, the mixture was cooled to room temperature, added to 20 mL of water, and extracted with dichloromethane (20 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to dryness, yielding 0.62 g of a yellow solid (66.2% yield).

[0077] Step 4: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-1)

[0078] C1 (0.50 g, 0.75 mmol) and trifluoroacetic acid (2 mL) were added to dry dichloromethane (10 mL), and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the solution was evaporated to dryness under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (26:1, v / v) as the eluent to give 0.22 g of a yellow solid, with a yield of 51.3%.

[0079] MS (ESI) m / z (%): 564.07 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.54(s,1H),8.48(d,J=9.5Hz,2H),8.29(s,1H),7.83(d,J=8.2Hz,2H),7.62(t,J=8.0Hz,1H), 7.40(d,J=25.0Hz,3H),7.20(s,1H),7.03(d,J=8.2Hz,1H),3.81(s,3H),3.44(p,J=6.9Hz,1H),1.15(d,J=6.9Hz,6H).

[0080] According to the synthesis method of Example 1, 2-fluoro-5-nitropyridine or 4-fluoronitrobenzene with different R1 substitutions (2-fluoro-5-nitropyridine, 2-nitro-4-bromo-5-fluoroanisole, 3-chloro-4-fluoronitrobenzene, 4-fluoronitrobenzene, 2-fluoro-5-nitroanisole) were used as raw materials to prepare B2 to B6 through synthetic route 3. Then, they were reacted with A1 through substitution and deprotection reactions to obtain compounds I-2 to I-6 of Examples 2 to 6.

[0081] Example 2: N 2-(6-((5-amino-1,3,4-thiadiazol-2-yl)thio)pyridin-3-yl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-2)

[0082] MS (ESI) m / z (%): 557.04 [M+Na] + ; 1 H NMR (400MHz, DMSO-d6) δ9.84(s,1H),9.50(s,1H),8.73(s,1H),8.48(d,J=8.5Hz,1H),8.34(s,1H),8.02(d,J=8.6Hz,1H),7.87(d,J=7 .8Hz,1H),7.75(t,J=7.9Hz,1H),7.55(s,2H),7.42(t,J=7.8Hz,1H),7.28(d,J=8.7Hz,1H),3.51–3.43(m,1H),1.16(d,J=6.7Hz,6H).

[0083] Example 3: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-5-bromo-2-methoxyphenyl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-3)

[0084] 1 H NMR (400MHz, DMSO-d6) δ9.52(s,1H),8.51(s,1H),8.45(d,J=8.5Hz,1H),8.36(s,1H),8.26(s,1H),7.85(d,J=8.0Hz,1H),7.7 2(t,J=8.0Hz,1H),7.48(s,2H),7.40(t,J=7.9Hz,1H),7.18(s,1H),3.79(s,3H),3.47(d,J=6.7Hz,1H),1.16(d,J=6.8Hz,6H).

[0085] Example 4: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-3-chlorophenyl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-4)

[0086] MS (ESI) m / z (%): 568.02 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ9.98(s,1H),9.47(s,1H),8.41(d,J=28.5Hz,2H),8.03(s,1H),7.88(d,J=7.7Hz,1H),7. 79(t,J=7.6Hz,1H),7.53(d,J=8.4Hz,1H),7.43(d,J=15.1Hz,4H),3.17(d,J=5.1Hz,1H),1.16(d,J=6.7Hz,6H).

[0087] Example 5: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)phenyl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-5)

[0088] MS (ESI) m / z (%): 534.06 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.80(s,1H),9.49(s,1H),8.53(d,J=8.4Hz,1H),8.34(s,1H),7.87(d,J=7.9Hz,1H),7.77(t, J=8.0Hz,1H),7.68(d,J=8.2Hz,2H),7.41(d,J=8.3Hz,3H),7.36(s,2H),3.47(d,J=6.9Hz,1H),1.16(d,J=6.8Hz,6H).

[0089] Example 6: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-3-methoxyphenyl)-5-chloro-N 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-6)

[0090] MS (ESI) m / z (%): 564.07 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ9.79(s,1H),9.49(s,1H),8.53(s,1H),8.36(s,1H),7.86(d,J=7.8Hz,1H),7.74(t,J=7.9Hz,1H) ,7.42(d,J=16.9Hz,2H),7.31(d,J=8.1Hz,2H),7.24(s,2H),3.64(s,3H),3.49(d,J=11.8Hz,1H),1.17(d,J=6.6Hz,6H).

[0091] According to the synthesis method of Example 1, 2,4,5-trichloropyrimidine and anilines with different R2 substitutions (2-methanesulfonylaniline, 2-amino-N-methylbenzamide, 5-aminoindan) were used to prepare A7 to A9 via synthetic route 2. Then, the compounds I-7 to I-9 of Examples 7 to 9 were obtained by substitution and deprotection reactions with B1.

[0092] Example 7: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N 4 -(2-(methanesulfonyl)phenyl)pyrimidine-2,4-diamine (I-7)

[0093] MS (ESI) m / z (%): 536.04 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.39(s,1H),8.38(d,J=7.7Hz,2H),8.28(s,1H),7.91(d,J=8.0Hz,1H),7.82(d,J=8.4Hz,1H),7. 62(t,J=8.1Hz,1H),7.41(d,J=20.9Hz,3H),7.19(d,J=2.1Hz,1H),6.99(dd,J=8.3,2.1Hz,1H),3.82(s,3H),3.26(s,3H).

[0094] Example 8: 2-((2-((4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)-N-methylbenzamide (I-8)

[0095] MS (ESI) m / z (%): 515.08 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ11.62(s,1H),8.76(d,J=5.0Hz,1H),8.55(d,J=8.4Hz,1H),8.32(s,1H),8.22(s,1H),7.94(d,J=8.3Hz,1H),7.7 4(d,J=7.9Hz,1H),7.38(d,J=20.5Hz,3H),7.21(s,1H),7.13(t,J=7.6Hz,1H),7.07(d,J=8.6Hz,1H),3.83(s,3H),2.80(d,J=4.4Hz,3H).

[0096] Example 9: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N 4 -(2,3-dihydro-1H-inden-5-yl)pyrimidine-2,4-diamine (I-9)

[0097] MS (ESI) m / z (%): 498.09 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.87(s,1H),8.12(s,1H),7.99(d,J=8.5Hz,1H),7.86(s,1H),7.41(d,J =16.1Hz,3H),7.28–7.12(m,3H),6.82(s,1H),3.84(s,3H),2.82(d,J=19.4Hz,4H),2.04(s,2H).

[0098] Example 10: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-methylpiperidin-1-yl)acetamide (I-10)

[0099] I-1 was prepared using synthetic route 1, and then I-10 was obtained through a two-step substitution reaction. The specific steps of the two-step substitution reaction are as follows:

[0100] Step 1: 2-Chloro-N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)acetamide (intermediate E1)

[0101] Compound I-1 (1.00 g, 1.77 mmol) and potassium carbonate (0.61 g, 4.43 mmol) were added to 15 mL of dry dichloromethane. Chloroacetyl chloride (1.24 g, 11.0 mmol) was then slowly added dropwise at 0 °C, and the mixture was stirred at 0 °C for 4 hours. After the reaction was complete, the reaction solution was cooled to room temperature, 20 mL of water was added, and the solution was adjusted to alkalinity with saturated potassium carbonate solution. The mixture was extracted with dichloromethane (20 mL × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure. 0.86 g of a yellowish-brown solid was obtained, with a yield of 75.8%.

[0102] Step 2: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-methylpiperidin-1-yl)acetamide (I-10)

[0103] Intermediate E1 (0.10 g, 0.16 mmol) and potassium carbonate (0.07 g, 0.47 mmol) were added to dry tetrahydrofuran (5 mL). 4-methylpiperidine (0.03 g, 0.31 mmol) was then slowly added dropwise at 0 °C. The reaction was carried out under nitrogen atmosphere and stirred at room temperature for 4 hours. After the reaction was complete, the reaction solution was cooled to room temperature, added to water, and extracted with dichloromethane (10 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane / methanol (25:1) as eluent to give 0.06 g of a yellow solid, with a yield of 57.3%.

[0104] MS (ESI) m / z (%): 703.17 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.56 (s, 1H), 8.49 (d, J = 8.4Hz, 1H), 8.40 (d, J = 8.5Hz, 1H), 8.20 (s,1H),7.94(d,J=7.9Hz,1H),7.70(d,J=11.1Hz,2H),7.31(t,J=7.6Hz,1H),7.21(d, J=8.5Hz,1H),7.15(s,1H),3.92(s,3H),3.17(s,2H),2.79(d,J=11.2Hz,2H),2.25(t ,J=11.5Hz,2H),1.66(d,J=12.6Hz,3H),1.32(d,J=6.9Hz,8H),0.96(d,J=6.0Hz,3H).

[0105] According to the synthesis method of Example 10, E1 was reacted with different small molecule fatty amines through substitution reactions to obtain compounds I-11 to I-22 of Examples 11 to 22.

[0106] Example 11: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-methylpiperazin-1-yl)acetamide (I-11)

[0107] MS (ESI) m / z (%): 704.17 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.57(s,1H),8.45(dd,J=32.7,8.4Hz,2H),8.20(s,1H),7.94(d,J=7.9Hz,1H),7.70(d,J=10.9Hz,2H),7 .31(d,J=7.6Hz,1H),7.23–7.12(m,2H),3.92(s,3H),3.23(s,3H),2.59(d,J=42.6Hz,8H),2.34(s,3H),1.32(d,J=6.8Hz,6H).

[0108] Example 12: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-(dimethylamino)piperidin-1-yl)acetamide (I-12)

[0109] MS (ESI) m / z (%): 732.20 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.57 (s, 1H), 8.45 (dd, J = 33.3, 8.3Hz, 2H), 8.20 (s, 1H) ),7.94(d,J=7.9Hz,1H),7.70(d,J=10.3Hz,2H),7.31(d,J=7.6Hz,1H),7.23 –7.13(m,2H),3.92(s,3H),3.23(d,J=9.8Hz,3H),2.44(s,8H),2.32(t,J=11 .8Hz,3H),1.94(d,J=12.1Hz,2H),1.75–1.64(m,2H),1.32(d,J=6.9Hz,6H).

[0110] Example 13: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-ethylpiperazin-1-yl)acetamide (I-13)

[0111] MS(ESI) m / z (%): 718.18 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.57(s,1H),8.45(dd,J=32.6,8.4Hz,2H),8.20(s,1H),7.94(d,J=7.9Hz,1H),7.70(d,J=12.1Hz,2H),7.30(t,J=7.6Hz,1H),7.2 1(d,J=8.5Hz,1H),7.15(s,1H),3.92(s,3H),3.23(s,3H),2.62(d,J=30.5H z, 8H), 2.49 (t, J = 7.2Hz, 2H), 1.32 (d, J = 6.8Hz, 6H), 1.11 (t, J = 7.2Hz, 3H).

[0112] Example 14: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(dimethylamino)acetamide (I-14)

[0113] MS (ESI) m / z (%): 671.11 [M+Na] + ; 1 H NMR(400MHz, CDCl3)δ9.56(s,1H),8.45(dd,J=34.7,8.4Hz,2H),8.20(s,1H),7.94(d,J=7.9Hz,1H),7.70(d,J=10.0Hz,2H),7 .31(t,J=7.6Hz,1H),7.24–7.13(m,2H),3.92(s,3H),3.24(p,J=6.8Hz,1H),3.16(s,2H),2.36(s,6H),1.32(d,J=6.8Hz,6H).

[0114] Example 15: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-((3-hydroxypropyl)amino)acetamide (I-15)

[0115] MS (ESI) m / z (%): 679.13 [M+H] + .

[0116] Example 16: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-(2-hydroxyethyl)piperazin-1-yl)acetamide (I-16)

[0117] MS (ESI) m / z (%): 734.18 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.57(s,1H),8.45(dd,J=31.9,8.4Hz,2H),8.20(s,1H),7.94(d,J=7.9Hz,1H),7.70(d,J=12.3Hz,2H),7.31(t,J=7.5H z,1H),7.23–7.13(m,2H),3.92(s,3H),3.66(t,J=5.3Hz,2H),3.24(s,3H),2.65(d,J=11.7Hz,8H),2.26–1.89(m,2H),1.32(d,J=6.9Hz,6H).

[0118] Example 17: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-hydroxypiperidin-1-yl)acetamide (I-17)

[0119] MS(ESI)m / z(%):705.15[M+H]+; 1 H NMR (400MHz, CDCl3) δ9.56 (s, 1H), 8.49 (d, J = 8.4Hz, 1H), 8.41 (d, J = 8.5Hz, 1H), 8.20(s,1H),7.94(d,J=8.0Hz,1H),7.74–7.67(m,2H),7.31(t,J=7.7Hz,1H),7. 21(d,J=8.5Hz,1H),7.15(s,1H),3.92(s,3H),3.81(s,1H),3.22(s,3H),2.80(s ,2H),2.46–2.40(m,2H),1.93(s,2H),1.71–1.64(m,2H),1.32(d,J=6.8Hz,6H).

[0120] Example 18: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(pyrrolidine-1-yl)acetamide (I-18)

[0121] MS (ESI) m / z (%): 675.14 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.56(s,1H),8.49(d,J=8.3Hz,1H),8.40(d,J=8.6Hz,1H),8.20(s,1H),7.94(d,J=7.9Hz,1H),7.74–7.64(m,2H),7.31(t, J=7.5Hz,1H),7.21(d,J=8.4Hz,1H),7.15(s,1H),3.92(s,3H),3.37(s, 2H),3.27–3.18(m,1H),2.68(s,4H),1.86(s,4H),1.32(d,J=6.8Hz,6H).

[0122] Example 19: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(diethylamino)acetamide (I-19)

[0123] MS (ESI) m / z (%): 677.16 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.56(s,1H),8.44(dd,J=36.1,8.4Hz,2H),8.20(s,1H),7.94(d,J=7.8Hz,1H),7.71(s,2H),7.30( s,1H),7.24–7.12(m,2H),3.92(s,3H),3.23(s,3H),2.63(q,J=7.2Hz,4H),1.32(d,J=6.9Hz,6H),1.06(t,J=7.1Hz,6H).

[0124] Example 20: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(3-hydroxyazacyclobutane-1-yl)acetamide (I-20)

[0125] MS (ESI) m / z (%): 677.12 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.57(s,1H),8.45(dd,J=34.0,8.4Hz,2H),8.20(s,1H),7.93(d,J=8.0Hz,1H),7.70(d,J=10.8Hz,2H),7.31( d,J=7.9Hz,1H),7.23–7.08(m,2H),4.51(s,1H),3.91(s,3H),3.79(s,2H),3.38(s,2H),3.31–3.08(m,3H),1.32(d,J=6.8Hz,6H).

[0126] Example 21: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-thiomorpholinylacetamide (I-21)

[0127] MS(ESI) m / z (%): 707.11 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.57(s,1H),8.49(d,J=8.4Hz,1H),8.41(d,J=8.5Hz,1H),8.20(s,1H),7.94(d,J=7.9Hz,1H),7.75–7.66(m,2H),7.32(d ,J=7.6Hz,1H),7.21(d,J=8.5Hz,1H),7.15(s,1H),3.92(s,3H),3.22(s,3H),2.84(t,J=4.7Hz,4H),2.78–2.71(m,4H),1.32(d,J=6.8Hz,6H).

[0128] Example 22: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(3-(dimethylamino)azacyclobutane-1-yl)acetamide (I-22)

[0129] MS (ESI) m / z (%): 704.17 [M+H] + ; 1H NMR (400MHz, CDCl3) δ9.56 (s, 1H), 8.44 (dd, J = 34.3, 8.4Hz, 2H), 8.20 (s, 1H) ,7.93(d,J=7.9Hz,1H),7.70(d,J=10.7Hz,2H),7.31(d,J=7.7Hz,1H),7.23– 7.13(m,2H),3.91(s,3H),3.71(s,1H),3.64(d,J=6.7Hz,2H),3.35(s,2H),3 .13(t,J=6.8Hz,2H),2.96–2.88(m,1H),2.12(s,6H),1.32(d,J=7.0Hz,6H).

[0130] According to the synthesis method of Example 10, 2,4,5-trichloropyrimidine and methanesulfonyl-substituted aniline were used to prepare A via synthetic route 2. 23 Then, through substitution, deprotection, and substitution reactions with B1, E is obtained. 23 E 23 Compounds I-23 to I-25 of Examples 23 to 25 were obtained by substitution reactions with different small molecule fatty amines.

[0131] Example 23: N-(5-((4-((5-chloro-4-((2-(methanesulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-methylpiperazin-1-yl)acetamide (I-23)

[0132] MS (ESI) m / z (%): 676.14 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.28(s,1H),8.42(dd,J=16.9,8.4Hz,2H),8.22(s,1H),8.02(d,J=7.9Hz,1H),7.71(dd,J=15.6,7.3Hz ,2H),7.33(t,J=7.8Hz,1H),7.21–7.10(m,2H),3.91(s,3H),3.22(s,2H),3.10(s,3H),2.57(d,J=46.5Hz,8H),2.32(s,3H).

[0133] Example 24: N-(5-((4-((5-chloro-4-((2-(methanesulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(dimethylamino)acetamide (I-24)

[0134] MS (ESI) m / z (%): 643.08 [M+Na] + . 1 H NMR(400MHz, CDCl3)δ9.28(s,1H),8.42(dd,J=18.4,8.4Hz,2H),8.22(s,1H),8.02(d,J=7.9Hz,1H),7.77– 7.66(m,2H),7.33(t,J=7.7Hz,1H),7.23–7.12(m,2H),3.91(s,3H),3.16(s,2H),3.10(s,3H),2.36(s,6H).

[0135] Example 25: N-(5-((4-((5-chloro-4-((2-(methanesulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(4-methylpiperidin-1-yl)acetamide. (I-25)

[0136] MS(ESI) m / z: 675.14 [M+H] + .

[0137] According to the synthesis method of Example 10, B was prepared via synthetic route 3 using 2-fluoro-5-nitropyridine or 4-fluoronitrobenzene with different R1 substitutions as raw materials. 26 ~B 32 Then, through substitution, deprotection, and substitution reactions with A1, E is obtained. 26-32 E 26-32 Compounds I-26 to I-32 of Examples 26 to 32 were obtained by substitution reactions with different small molecule fatty amines.

[0138] Example 26: N-(5-((5-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyridin-2-yl)thio)-1,3,4-thiadiazol-2-yl)-2-(dimethylamino)acetamide (I-26)

[0139] MS(ESI) m / z: 620.11 [M+H] + .

[0140] Example 27: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-2-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(dimethylamino)acetamide (I-27)

[0141] MS(ESI) m / z: 649.12 [M+H] +;(400MHz, CDCl3)δ9.62(s,1H),8.50(d,J=8.3Hz,1H),8.21(s,1H),7.92(d,J=7.9Hz,1H),7.66(t,J=8.0Hz,1H),7.51(d, J=8.4Hz,1H),7.38(s,1H),7.11(d,J=8.4Hz,1H),3.72(s,3H),3.21(d,J=34.4Hz,3H),2.37(s,6H),1.32(d,J=6.9Hz,6H).

[0142] Example 28: N-(5-((2-chloro-4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)phenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(dimethylamino)acetamide (I-28)

[0143] MS(ESI) m / z: 653.07 [M+H] + .

[0144] Example 29: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)phenyl)thio)-1,3,4-thiadiazol-2-yl)-2-(dimethylamino)acetamide (I-29)

[0145] MS(ESI) m / z: 619.11 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.88(s,1H),9.49(s,1H),8.53(d,J=8.0Hz,1H),8.35(s,1H),7.86(d,J=7.9Hz,1H),7. 81–7.72(m,3H),7.53(d,J=8.1Hz,2H),7.41(s,1H),3.45(q,J=7.0Hz,2H),2.30(s,6H),1.16(d,J=6.6Hz,6H).

[0146] Example 30: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)phenyl)thio)-1,3,4-thiadiazol-2-yl)-2-((3-hydroxypropyl)amino)acetamide (I-30)

[0147] MS(ESI) m / z: 649.12 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),9.48(s,1H),8.53(d,J=8.4Hz,1H),8.33(s,1H),7.84(d,J=7.9Hz,1H),7.75(t,J=7.9Hz ,1H),7.68(d,J=8.3Hz,2H),7.40(dd,J=20.4,7.8Hz,3H),3.57–3.38(m,6H),2.83(s,2H),1.63(s,2H),1.16(d,J=6.8Hz,6H).

[0148] Example 31: N-(5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-2-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)-2-((3-hydroxypropyl)amino)acetamide (I-31)

[0149] MS(ESI) m / z: 679.14 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.81(s,1H),9.49(s,1H),8.54(d,J=8.4Hz,1H),8.37(s,1H),7.85(d,J=7.8Hz,1H),7.75(t,J=8.0Hz,1H),7.46(s,1H),7.3 9(d,J=7.7Hz,1H),7.32(s,2H),3.61(s,3H),3.51(s,2H),3.44(t,J=6.5H z, 3H), 2.76 (d, J = 7.4Hz, 2H), 1.66 (t, J = 7.0Hz, 2H), 1.17 (d, J = 6.8Hz, 6H).

[0150] Example 32: N-(5-((5-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyridin-2-yl)thio)-1,3,4-thiadiazol-2-yl)-2-((3-hydroxypropyl)amino)acetamide (I-32)

[0151] MS(ESI) m / z: 650.12 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ9.83(s,1H),9.48(s,1H),8.72(s,1H),8.52–8.42(m,1H),8.32(s,1H),7.97(d,J=8.7Hz,1H),7.83(d,J=7.9Hz, 1H),7.72(t,J=8.0Hz,1H),7.35(s,1H),7.23(d,J=8.7Hz,1H),3.49(d,J=31.3Hz,6H),2.90(s,2H),1.69(s,2H),1.14(d,J=6.7Hz,6H).

[0152] Biological activity studies of compounds I-1 to I-32 provided by this invention.

[0153] This invention provides a study on the inhibitory activity of compounds I-1 to I-32 against ALK and ALK mutants.

[0154] Compounds I-1 to I-32 provided by this invention were subjected to inhibition of ALK and ALK. L1196M ALK G1202R Activity screening. The specific procedure is as follows:

[0155] 1. Prepare compounds for testing kinases.

[0156] 1) Dilute the compound to the final desired maximum inhibitor concentration 100-fold with 100% dimethyl sulfoxide. Transfer 100 μL of the compound dilution to one well of a 96-well plate. For example, if the desired maximum inhibitor concentration is 0.1 μM, prepare a 10 μM solution of the compound in DMSO in this step.

[0157] 2) Add 100 μL of 100% DMSO solution to two wells of the same 96-well plate, one for the compound-free control and one for the enzyme-free control. Label this plate as the source plate.

[0158] 3) Transfer 40 μL of the compound from the source plate to a new 384-well Echo plate as an intermediate plate.

[0159] 4) Transfer 100 nL of the compound per well in the 384-well Echo plate to the 384-well detection plate via reflux.

[0160] 2. Kinase response.

[0161] 1) Except for the control wells without enzyme (add 5 μL of 1x kinase buffer), add 5 μL of kinase solution to each well of the test plate.

[0162] 2) Prepare substrate solutions of the substrate and adenosine triphosphate (ATP) in 1x kinase reaction buffer, with the final concentration of each reagent being twice the concentration required for the assay.

[0163] 3) Add 5 μL of substrate solution to each well of the detection plate.

[0164] 4) Incubate the 384-well detection plate at room temperature for 30 or 60 minutes.

[0165] 5) Prepare kinase quenching buffer and antibody detection solutions. The final concentration of each reagent in Lance detection buffer should be twice the required concentration.

[0166] 6) Add 10 μL of detection solution and let stand at room temperature for 60 minutes.

[0167] 3. Curve Fitting

[0168] 1) Copy the Lance signal ratio (665 nm / 615 nm) from the Envision program.

[0169] 2) Convert the ratio value to a suppression percentage value.

[0170] a. Suppression percentage = (maximum value - sample Lance signal ratio) / (maximum value - minimum value) * 100.

[0171] b. "Minimum" indicates the ratio of enzyme-free control, and "maximum" indicates the ratio of DMSO control.

[0172] 3) The data is displayed in MS Excel, and the IC curve is fitted using XLFit Excel add-in version 5.4.0.8. 50 The calculation formula is: Y = Bottom + (Top - Bottom) / (1 + (IC) 50 / X)^HillSlope).

[0173] The results of the kinase inhibition rate test of compounds I-1 to I-32 against ALK are shown in the table below.

[0174]

[0175]

[0176] Compounds with good ALK kinase inhibitory activity were tested for their ALK mutant kinase inhibitory activity. The inhibitory effects of I-1, I-3, I-6, I-7, I-8, I-14, and I-18 on ALK and ALK kinase were assessed. L1196M and ALK G1202R The results of the kinase inhibitory activity assay are shown in the table below, in nM.

[0177]

[0178]

[0179] The test results above clearly show that compounds I-1 to I-32, which are to be protected by this invention, are effective against ALK and ALK2. L1196M mutant, ALK G1202R The mutants exhibit significant inhibitory activity, with compound I-1 showing significantly better inhibitory activity than the control drug Ceritinib. This demonstrates that the compounds provided by this invention can be used to prepare ALK and ALK mutant inhibitors.

[0180] In this invention, compounds of general formula I can be administered alone, but are usually given in mixture with a pharmaceutical carrier, the choice of which depends on the desired route of administration and standard pharmaceutical practice.

[0181] Although the invention has been described with reference to specific embodiments, modifications and equivalent variations will be apparent to those skilled in the art, and are all included within the scope of the invention.

Claims

1. A 2,5-disubstituted thiadiazole compound containing a pyrimidine ring as shown in Formula I, or a pharmaceutically acceptable salt thereof, characterized in that, Structural formula: In Formula I: X is selected from C and N; R1 is selected from H, C1-C6 alkoxy, or halogen; there may be one or more R1s. A is a phenyl group; R2 is selected from H, C1-C3 alkylsulfonyl, C1-C3 alkylamide, or R2 together with the carbon atom on A to form a 4-6 membered cycloalkyl group; R3 is selected from amino groups, ; R4 and R5 may be the same or different, and are independently selected from H, C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, or R4 and R5 together with the nitrogen atom to which they are attached to form a 4-6 membered heterocyclic group, wherein the heterocyclic group, in addition to the nitrogen atom attached to R4 and R5, optionally contains 0-1 heteroatom selected from N, O and S, and the heterocyclic group is optionally substituted by 0-1 R6; R6 is selected from H, hydroxyl, C1-C6 alkyl, and C1-C6 alkylamino.

2. The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring according to claim 1, characterized in that, It is any one of the compounds I-1 to I-32 shown below: 。 3. A pharmaceutical composition, characterized in that, The active ingredient is a 2,5-disubstituted thiadiazole compound containing a pyrimidine ring as described in any one of claims 1-2, or a pharmaceutically acceptable salt thereof, which is mixed with a pharmaceutically acceptable excipient. The dosage forms include injections, tablets, capsules, aerosols, suppositories, films, pellets, topical liniments, and ointments.

4. The use of the 2,5-disubstituted thiadiazole compound containing a pyrimidine ring according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 3, in the preparation of a drug for inhibiting ALK mutants, characterized in that, The ALK mutant is one or more of G1202R, L1196M, G1269A, L1198F, and F1174L.

5. The method for preparing the 2,5-disubstituted thiadiazole compound containing a pyrimidine ring as described in claim 2, characterized in that, The synthetic route includes the following steps: Synthesis Route 1: In the process of preparing compounds I-10 to I-32 by reacting intermediate E with aliphatic amines, the aliphatic amines used are selected from the following compounds: Synthetic route 2: Preparation of intermediate A Synthetic route 3: Preparation of intermediate B In synthetic routes 1-3, R1, R2, R3, A, and X are the corresponding functional groups at the corresponding positions of compounds I-1 to I-32.

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