Pyrimidine ring-containing 2, 5-disubstituted thiadiazole compound as well as preparation method and application thereof

Abstract

The invention discloses a pyrimidine ring-containing 2, 5-disubstituted thiadiazole compound as well as a preparation method and application thereof, belongs to the technical field of medicines, and provides a pyrimidine ring-containing 2, 5-disubstituted thiadiazole compound with a structural formula as shown in the specification or a pharmaceutically acceptable salt thereof and a preparation method of the pyrimidine ring-containing 2, 5-disubstituted thiadiazole compound. The invention also relates to application of the compound or the pharmaceutically acceptable salt thereof or the pharmaceutical composition containing the compound in preparation of drugs for treating diseases related to abnormal expression of anaplastic lymphoma kinase. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a novel 2,5-disubstituted thiadiazole compound containing a pyrimidine ring and a preparation method thereof, and use of the compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound in preparing a drug for treating a disease associated with abnormal expression of anaplastic lymphoma kinase. Background Art

[0002] Anaplastic lymphoma kinase (ALK) is a highly conserved transmembrane receptor tyrosine kinase that belongs to the insulin receptor (IR) kinase superfamily. Structurally, ALK consists of an N-terminal extracellular domain, a hydrophobic single-pass transmembrane region, and an intracellular kinase domain. When ALKAL protein (the endogenous ligand of ALK) binds to its extracellular domain, ALK is activated, leading to dimerization and autophosphorylation, which results in dysregulation of cell proliferation and survival. In 1994, Morris et al. discovered NPM1-ALK gene rearrangement mutations in patients with anaplastic large cell lymphoma. It was not until 2007 that 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 cell signaling pathways (such as RAS-MAPK, PI3K-AKT, and JAK-STAT).

[0003] According to statistics, since the first report of EML4-ALK gene rearrangement mutation in NSCLC patients, about 3%-7% of NSCLC patients have ALK rearrangement. Although ALK rearrangement accounts for a small proportion, the large number of NSCLC cases has resulted in about 40,000 new cases of this type of patients worldwide each year. At present, ALK inhibitors have maintained a high level of development and research in the targeted treatment of NSCLC. With the continuous clarification of the pathogenesis and drug resistance mechanisms, new ALK inhibitors continue to emerge, among which small molecule ALK inhibitors have undergone four generations of drug changes.

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

[0005] Although the fourth-generation ALK inhibitors have overcome some ALK compound mutations, various drug-resistant mutations have emerged in response to different inhibitors. At present, the treatment strategies of existing ALK inhibitors are not enough to meet the clinical treatment needs. In view of the susceptibility of ALK active site mutations and other issues, there is still an urgent need to develop new ALK inhibitors that can overcome drug-resistant mutations for the treatment of non-small cell lung cancer. Summary of the invention

[0006] The primary purpose of the present invention is to provide a novel pyrimidine ring-containing 2,5-disubstituted thiadiazole compound as shown in general formula I and a preparation method thereof. Also provided is the use of the compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound in the preparation of a drug for treating a disease associated with anaplastic lymphoma kinase mutation.

[0007]

[0008] in:

[0009] X is selected from C and N;

[0010] R1 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen, halogenated (C1-C6) alkyl, nitro; R1 can be 1 or more;

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

[0012] R2 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen, (C1-C6) alkylsulfonyl, (C1-C6) alkylamido; or R2 and the carbon atoms on A together 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 are the same or different and are independently selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, hydroxyl, and 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 to which R4 and R5 are attached, optionally contains 0-1 heteroatoms selected from N, O and S, and the heterocyclic group is optionally substituted by 0-1 R6;

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

[0017] Furthermore, in the general formula I:

[0018] R1 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen; R1 can be 1 or more;

[0019] A is selected from 5-6 membered 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) alkylamido; or R2 and the carbon atom on A together form a 4-6 membered cycloalkyl;

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

[0022] R4 and R5 are the same or different and are independently selected from H, (C1-C6) alkyl, hydroxyl, and (C1-C6) alkyl substituted with hydroxyl; 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 to which R4 and R5 are attached, 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, hydroxy, amino, (C1-C6) alkyl, (C1-C6) alkyl substituted with hydroxy or halogen or amino or cyano or carboxyl, (C1-C6) alkylamino, di(C1-C6) alkylamino.

[0024] Furthermore, in the general formula I:

[0025] R1 is selected from H, (C1-C6) alkoxy, halogen; R1 can be 1 or more;

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

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

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

[0029] R4 and R5 are the same or different and are independently selected from H, (C1-C6) alkyl, and hydroxy-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 to which R4 and R5 are attached, 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, hydroxy, (C1-C6) alkyl, hydroxy-substituted (C1-C6) alkyl, di(C1-C6) alkylamino.

[0031] Furthermore, in the general formula I:

[0032] R1 is selected from H, methoxy, Cl; R1 can be 1 or more;

[0033] A is phenyl;

[0034] R2 is selected from H, isopropylsulfonyl, methylamido; or R2 and the carbon atom on A together form cyclopentane;

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

[0036]

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

[0038]

[0039]

[0040]

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

[0042] In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to a straight chain or branched alkyl; "heterocyclic group" refers to a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S.

[0043] The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring represented by the general formula I of the present invention or a pharmaceutically acceptable salt thereof is used as an active ingredient, mixed with a pharmaceutically acceptable excipient to prepare a composition, and prepared into a clinically acceptable dosage form, wherein the excipient refers to a diluent, adjuvant or carrier that can be used in the pharmaceutical field. The dosage form includes injections, tablets, capsules, aerosols, suppositories, films, pills, external liniments and ointments commonly used in clinical practice.

[0044] The 2,5-disubstituted thiadiazole compounds containing a pyrimidine ring of the present invention have the function of significantly inhibiting ALK kinase, especially ALK which is closely related to the generation of drug resistance. G1202 ALK L1196M The mutation has a potent 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 for inhibiting ALK mutants, wherein the ALK mutant is one or more of G1202R, L1196M, G1269A, L1198F, and F1174L.

[0046] The invention relates to use of the pyrimidine ring-containing 2,5-disubstituted thiadiazole compound or a pharmaceutical composition comprising the compound in the preparation of a drug 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 ring-containing 2,5-disubstituted thiadiazole compounds of the present invention.

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

[0049] Synthetic route 1:

[0050]

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

[0052]

[0053] Preparation method of intermediate A such as synthetic route;

[0054] Synthesis route 2:

[0055]

[0056] Preparation method of intermediate B such as synthetic route;

[0057] Synthetic route 3:

[0058]

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

[0060] Beneficial effects of the present invention:

[0061] The compound of the present invention has a novel chemical structure and has a high inhibitory activity on ALK mutant kinase in in vitro studies, and can be used for the treatment and prevention of various diseases such as cancer. DETAILED DESCRIPTION

[0062] In the following examples, methods for preparing some of the compounds of Formula I are provided. It should be understood that the following methods and other methods known to those of ordinary skill in the art can be applied to the preparation of all compounds described in the present invention. The examples are intended to illustrate rather than limit the scope of the present invention.

[0063] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents and instruments used, they are all conventional reagent products that can be purchased commercially.

[0064] Example 1: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N-(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl) 4 -(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine (I-1) was prepared by the method of synthetic route 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 dry N,N-dimethylformamide (15 mL), and stirred at 0°C for 0.5 hours in a nitrogen system. Then 2,4,5-trichloropyrimidine (2.00 g, 11.0 mmol) dissolved in dry N,N-dimethylformamide was slowly added, and the reaction was carried out at room temperature for 12 hours. After the reaction was completed, the reaction solution was added to cold water, and the solid was precipitated. It was filtered and the filter cake was dried to obtain 3.30 g of a yellow-brown solid with a yield of 86.9%.

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

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

[0070] 5-Fluoro-2-nitroanisole (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 stirred at 50°C for 4 hours. After the reaction was completed, the reaction solution was added to cold water to precipitate solids, which were filtered by suction, and the filter cake was washed with cold water and dried to obtain 1.53 g of yellow solids with a yield of 92.0%.

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

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

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

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

[0075] Step 3: tert-Butyl (5-((4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)thio)-1,3,4-thiadiazol-2-yl)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), and the reaction solution was heated to 100° C. and stirred for 10 hours in a nitrogen system. After the reaction was completed, the reaction solution was cooled to room temperature, added to 20 mL of water, extracted with dichloromethane (20 mL×3), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure to obtain 0.62 g of a yellow solid with a yield of 66.2%.

[0077] Step 4: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N-(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl) 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 stirred at room temperature for 2 hours. After the reaction was completed, the solution was evaporated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column using dichloromethane / methanol (26:1, v / v) as eluent to obtain 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 as raw materials (2-fluoro-5-nitropyridine, 2-nitro-4-bromo-5-fluoroanisole, 3-chloro-4-fluoronitrobenzene, 4-fluoronitrobenzene, 2-fluoro-5-nitrobenzene) were used as raw materials through synthetic route 3 to prepare B2~B6, and then reacted with A1 through substitution and deprotection to obtain compounds I-2~I-6 of Examples 2~6.

[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-((5-amino-1,3,4-thiadiazol-2-yl)thio)-3-chlorophenyl)- 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-methylsulfonylaniline, 2-amino-N-methylbenzamide, 5-aminoindane) are prepared through synthesis route 2 to prepare A7~A9, and then reacted with B1 through substitution and deprotection reaction to obtain compounds I-7~I-9 of Examples 7~9.

[0092] Example 7: N 2 -(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl)-5-chloro-N-(4-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl) 4 -(2-(Methylsulfonyl)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)sulfur)-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-((5-amino-1,3,4-thiadiazol-2-yl)thio)-2-methoxyphenyl) 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] The method of synthetic route 1 is used to prepare I-1, and then I-10 is 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 dry dichloromethane (15 mL), and chloroacetyl chloride (1.24 g, 11.0 mmol) was slowly added dropwise at 0°C, and the mixture was stirred at 0°C for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature, added to 20 mL of water, and the solution was adjusted to alkalinity with a saturated potassium carbonate solution, 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 to obtain 0.86 g of a yellow-brown solid 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] Add intermediate E1 (0.10 g, 0.16 mmol) and potassium carbonate (0.07 g, 0.47 mmol) to dry tetrahydrofuran (5 mL), slowly add 4-methylpiperidine (0.03 g, 0.31 mmol) at 0 ° C, stir at room temperature in a nitrogen system for 4 hours. After the reaction is completed, the reaction solution is cooled to room temperature, added to water, extracted with dichloromethane (10 mL × 3), the organic layers are combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure to obtain a crude product. The crude product is purified by silica gel column with dichloromethane / methanol (25:1) as the eluent to obtain 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 is reacted with different small molecular fatty amines to obtain compounds I-11 to I-22 of Examples 11 to 22 through substitution reaction.

[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-(pyrrolidin-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-hydroxyazetidin-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-thiomorpholinoacetamide (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)azetidin-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, it reacts with B1 through substitution, deprotection, and substitution to obtain E 23 , E 23 Compounds I-23 to I-25 of Examples 23 to 25 were obtained by substitution reaction with different small molecular fatty amines.

[0131] Example 23: N-(5-((4-((5-chloro-4-((2-(methylsulfonyl)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-(methylsulfonyl)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-(methylsulfonyl)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, 2-fluoro-5-nitropyridine or 4-fluoronitrobenzene with different R1 substitutions was used as a raw material to prepare B by synthetic route 3 26 ~B 32 Then, it reacts with A1 through substitution, deprotection, and substitution to obtain E 26-32 , E 26-32 The compounds I-26 to I-32 of Examples 26 to 32 were obtained by substitution reaction with different small molecular fatty amines.

[0138] Example 26: N-(5-((5-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)pyridin-2-yl)sulfur)-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] The biological activities of compounds I-1 to I-32 provided by the present invention are studied.

[0153] The compounds I-1 to I-32 provided by the present invention are studied on the activity of inhibiting ALK and ALK mutants.

[0154] The compounds I-1 to I-32 provided by the present invention were tested for inhibition of ALK, ALK L1196M , ALK G1202R Activity screening. The specific operations are:

[0155] 1. Preparation of Test Kinase Compounds

[0156] 1) Dilute the compound to 100 times the final desired maximum inhibitor concentration in 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 compound DMSO solution in this step.

[0157] 2) Add 100 μL of 100% DMSO solution to two empty wells of the no-compound control and no-enzyme control in the same 96-well plate and mark this plate as the source plate.

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

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

[0160] 2. Kinase reaction.

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

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

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

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

[0165] 5) Prepare kinase quenching buffer and antibody detection solution, with the final concentration of each reagent in Lance detection buffer being twice the required concentration.

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

[0167] 3. Curve fitting

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

[0169] 2) Convert the ratio values ​​to inhibition percentage values.

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

[0171] b. "Minimum" indicates the ratio of the no-enzyme control, and "Maximum" indicates the ratio of the DMSO control.

[0172] 3) Data were displayed in MS Excel and IC curves were 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 test results of the kinase inhibition rate of compounds I-1 to I-32 against ALK are shown in the table below.

[0174]

[0175]

[0176] The compounds with good ALK kinase inhibition rate were tested for ALK mutant kinase inhibition activity. The inhibition of ALK, ALK L1196M ALK G1202R The kinase inhibitory activity test results are shown in the table below, in nM.

[0177]

[0178]

[0179] It can be clearly seen from the above test results that the compounds I-1 to I-32 to be protected by the present invention have an inhibitory effect on ALK, ALK L1196M Mutant, ALK G1202R The mutants have significant inhibitory activity, among which compound I-1 is significantly better than the inhibitory activity of the control drug Ceritinib. It can be seen that the compounds provided by the present invention can be used to prepare ALK and ALK mutant inhibitors.

[0180] The compounds of formula I of this invention may be administered alone but will generally be administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice.

[0181] Although the present invention has been described in terms of specific embodiments, modifications and equivalents will be apparent to those skilled in the art and are intended to be encompassed within the scope of the present 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: Where: X is selected from C and N; R1 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen, halogenated (C1-C6) alkyl, nitro; R1 can be 1 or more; A is selected from 5-6 membered aryl or heteroaryl, wherein the heteroaryl contains 1-3 heteroatoms selected from N, O and S; R2 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen, (C1-C6) alkylsulfonyl, (C1-C6) alkylamido; or R2 and the carbon atoms on A together form a 4-6 membered cycloalkyl or aryl group; R3 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen, cyano, nitro, hydroxyl, amino, R4 and R5 are the same or different and are independently selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, hydroxyl, and 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 to which R4 and R5 are attached, optionally contains 0-1 heteroatoms selected from N, O and S, and the heterocyclic group is optionally substituted by 0-1 R6; R6 is selected from H, halogen, cyano, nitro, hydroxyl, carboxyl, amino, (C1-C6)alkyl, (C1-C6)alkyl substituted with hydroxyl or halogen or amino or cyano or carboxyl, (C1-C6)alkoxy, (C1-C6)alkylamino, di(C1-C6)alkylamino, (C1-C6)alkylcarbonyl, (C1-C6)alkylsulfonyl, (C1-C6)alkylsulfonylamino, and (C1-C6)alkylacylamino.

2. The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: In Formula I: R1 is selected from H, (C1-C6) alkyl, (C1-C6) alkoxy, halogen; R1 can be 1 or more; A is selected from 5-6 membered aryl or heteroaryl, wherein the heteroaryl contains 0-1 heteroatom selected from N, O and S; R2 is selected from H, (C1-C6) alkylsulfonyl, (C1-C6) alkylamido; or R2 and the carbon atom on A together form a 4-6 membered cycloalkyl; R3 is selected from H, (C1-C6) alkyl, amino, R4 and R5 are the same or different and are independently selected from H, (C1-C6) alkyl, hydroxyl, and (C1-C6) alkyl substituted with hydroxyl; 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 to which R4 and R5 are attached, optionally contains 0-1 heteroatoms selected from N, O and S, and the heterocyclic group is optionally substituted by 0-1 R6; R6 is selected from H, hydroxy, amino, (C1-C6)alkyl, (C1-C6)alkyl, (C1-C6)alkyl substituted with hydroxy or halogen or amino or cyano or carboxyl, (C1-C6)alkylamino, di(C1-C6)alkylamino.

3. The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: In Formula I: R1 is selected from H, (C1-C6) alkoxy, halogen; R1 can be 1 or more; A is selected from 5-6 membered aryl; R2 is selected from H, (C1-C3) alkylsulfonyl, (C1-C3) alkylamido; or R2 and the carbon atom on A together form a 4-6 membered cycloalkyl; R3 is selected from H, methyl, amino, R4 and R5 are the same or different and are independently selected from H, (C1-C6) alkyl, and hydroxy-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 to which R4 and R5 are attached, optionally contains 0-1 heteroatoms selected from N, O and S, and the heterocyclic group is optionally substituted by 0-1 R6; R6 is selected from H, hydroxy, (C1-C6) alkyl, hydroxy-substituted (C1-C6) alkyl, di(C1-C6) alkylamino.

4. The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: In Formula I: R1 is selected from H, methoxy, Cl; R1 can be 1 or more; A is phenyl; R2 is selected from H, isopropylsulfonyl, methylamido; or R2 and the carbon atom on A together form cyclopentane; R3 is selected from H, methyl, amino and the following structures:

5. The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring or a pharmaceutically acceptable salt thereof according to claim 4, characterized in that: Any one of the compounds I-1 to I-32 shown below:

6. A pharmaceutical composition, characterized in that The 2,5-disubstituted thiadiazole compound containing a pyrimidine ring or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 is used as an active ingredient and mixed with a pharmaceutically acceptable excipient. The dosage forms include injections, tablets, capsules, aerosols, suppositories, films, pills, external liniments and ointments.

7. Use of the pyrimidine ring-containing 2,5-disubstituted thiadiazole compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 6 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.

8. Use of the pyrimidine ring-containing 2,5-disubstituted thiadiazole compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 6 in the preparation of a drug for treating cancer.

9. The use according to claim 8, characterized in that: The cancer is non-small cell lung cancer.

10. The method for preparing the pyrimidine ring-containing 2,5-disubstituted thiadiazole compound or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that: The synthetic route comprises the following steps: Synthetic route 1: In the process of preparing compounds I-10 to I-32 by reacting intermediate E with aliphatic amine, the aliphatic amine used is selected from the following compounds: Synthetic route 2: Preparation of intermediate A Synthetic route 3: Preparation of intermediate B R1, R2, R3, A, and X in the synthetic route 1-3 are all corresponding groups at the corresponding positions of compounds I-1 to I-32.

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