A class of SIRT5 protein inhibitors and uses thereof

Abstract

The present invention provides a class of SIRT5 protein inhibitors and their uses, relating to the field of pharmaceutical technology. The SIRT5 protein inhibitors of the present invention are compounds represented by Formula I, or stereoisomers, tautomers, chiral isomers, or salts thereof. The compounds of the present invention can be used to prepare SIRT5 protein inhibitors and drugs for treating cancer. The present invention provides more options for the development and application of small molecule SIRT5 inhibitors and anticancer drugs. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a class of SIRT5 protein inhibitors and uses thereof. Background Art

[0002] Silent Information Regulator 2 (SIRT) is a type of protein that depends on the coenzyme NAD + Atypical histone deacetylases that remove acyl groups on ε-N of histone or non-histone lysines, including seven members from SIRT1 to SIRT7. SIRT family proteins all have a Zn 2+ binding domain, a Rossmann fold domain and a catalytic core domain (i.e. substrate and NAD + Compared with other members of the SIRT family, SIRT5 has a larger lysine acyl binding pocket and a specific tyrosine (Tyr102) and arginine residue (Arg105) in the pocket. Therefore, in addition to weak deacetylation activity, it also has significant acidic acylation activities such as demalonylation, desuccinylation, and deglutamylation, and its catalytic efficiency is about 1000 times that of deacetylation. Numerous studies have found that SIRT5 not only regulates the deacetylation of proteins such as CPS1, UOX, and CytC, thereby promoting physiological functions such as the urea cycle; it also regulates the desuccinylation of proteins such as HMGCS2, ECHA, GLS, SOD1, and SHMT2, thereby regulating ketone body synthesis, fatty acid β-oxidation, autophagy and mitochondrial autophagy, ROS clearance, and serine metabolism. It also regulates the demalonylation of proteins such as GAPDH and ALDOB, thereby regulating glucose metabolism. Furthermore, it catalyzes the deglutamylation of G6PD, GLUD1, and CPS1 proteins, thereby protecting cells from oxidative damage and promoting glutamine metabolism. Given the important physiological roles of SIRT5, studies have shown that abnormal SIRT5 expression is closely associated with the occurrence and development of tumors, cardiovascular diseases, and neurological diseases, and is considered an effective target for drug development for these diseases.

[0003] As the role of SIRT5 in various diseases has gradually been revealed, SIRT5 inhibitors have been reported in recent years. However, only a few peptide analogs have shown good in vitro and in vivo activity, and the few non-peptide small molecule inhibitors have low inhibitory activity against SIRT5 and are non-specific inhibitors. In addition, because non-peptide small molecule inhibitors have advantages in terms of pharmacokinetics and other drugability, there is an urgent need to develop new, highly effective and specific SIRT5 small molecule inhibitors to provide candidate drug molecules for the development of drugs targeting SIRT5-related diseases. Summary of the Invention

[0004] The purpose of the present invention is to provide a class of SIRT5 protein inhibitors and uses thereof.

[0005] The present invention provides a compound represented by Formula I, or a stereoisomer, or a tautomer, or a chiral isomer, or a salt thereof:

[0006]

[0007] in,

[0008] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C8 alkyl, C1-C8 alkoxy; n is an integer selected from 1 to 5;

[0009] R2 is selected from hydrogen, substituted or unsubstituted C1-C8 alkyl; the substituent of the alkyl is selected from -OC(O)R7, -OC(O)OR7;

[0010] R7 is selected from hydrogen, C1-C8 alkyl;

[0011] R3 and R4 are independently selected from hydrogen and C1-C8 alkyl;

[0012] R5, R6 are independently selected from hydrogen, and R5 and R6 are not simultaneously selected from hydrogen or

[0013] R8 is selected from

[0014] X is selected from -O- or -NH-;

[0015] R9 is selected from hydrogen, -C(S)NHR 10 , substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted 5-10 membered aryl, substituted or unsubstituted 3-10 membered cycloalkyl, substituted or unsubstituted 3-10 membered heterocycloalkyl; the alkyl substituent is selected from -C(S)NHR 10 、-C(O)OR 10 , 5-10 membered aryl, 5-10 membered heteroaryl; the substituents of the cycloalkyl and heterocycloalkyl groups are selected from hydroxyl, substituted or unsubstituted 5-10 membered aryl; the substituents of the aryl groups are selected from C1-C8 alkyl, halogen, hydroxyl, C1-C8 alkoxy, 5-10 membered aryl; the heteroatom of the heterocycloalkyl or heteroaryl group is N, O or S, and the number of the heteroatoms is 1, 2 or 3;

[0016] R 10 is selected from hydrogen, substituted or unsubstituted C1-C8 alkyl; the substituent of the alkyl is selected from -C(O)OR 11 ;

[0017] R 11 Selected from C1 to C8 alkyl;

[0018] Ring A is selected from a substituted or unsubstituted 4- to 10-membered heterocycloalkyl group containing a nitrogen atom, wherein the heterocycloalkyl group is connected to the keto group via the nitrogen atom; and the substituent of the heterocycloalkyl group is selected from a 5- to 10-membered aryl group;

[0019] R2, R3, R4 are selected from hydrogen, R6 is selected from hydrogen, R5 is selected from R8 is selected from When x is selected from -O- and R9 is selected from ethyl, R1 does not form a

[0020] R2, R3, R4 are selected from hydrogen, R6 is selected from hydrogen, R5 is selected from R8 is selected from X is selected from -NH-, R9 is selected from 3-membered cycloalkyl or When R1 and the benzene ring do not form

[0021] Furthermore, the structure of the compound is shown in Formula II:

[0022]

[0023] in,

[0024] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C8 alkyl, C1-C8 alkoxy; n is an integer selected from 1 to 5;

[0025] R9 is selected from hydrogen, C1-C8 alkyl;

[0026] When R9 is selected from ethyl, R1 and the benzene ring do not form

[0027] Preferably, R9 is selected from hydrogen, C1-C3 alkyl;

[0028] More preferably, the structure of the compound is as shown in Formula IIa:

[0029]

[0030] in,

[0031] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C3 alkyl, C1-C3 alkoxy; n is selected from 1 or 2;

[0032] Alternatively, the structure of the compound is shown in Formula IIb:

[0033]

[0034] in,

[0035] a is the number of methylene groups, selected from integers of 0 to 2;

[0036] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C3 alkyl, C1-C3 alkoxy; n is selected from 1 or 2.

[0037] Furthermore, the structure of the compound is shown in Formula III:

[0038]

[0039] in,

[0040] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C8 alkyl, C1-C8 alkoxy; n is an integer selected from 1 to 5;

[0041] R2 is selected from hydrogen, substituted or unsubstituted C1-C8 alkyl; the substituent of the alkyl is selected from -OC(O)R7, -OC(O)OR7;

[0042] R7 is selected from C1-C8 alkyl;

[0043] m is an integer from 0 to 5;

[0044] R 12 is selected from hydrogen or a 5- to 10-membered aryl group;

[0045] Preferably, the structure of the compound is as shown in Formula IV:

[0046]

[0047] in,

[0048] R2 is selected from hydrogen, substituted or unsubstituted C1-C8 alkyl; the substituent of the alkyl is selected from -OC(O)R7, -OC(O)OR7;

[0049] R7 is selected from C1-C8 alkyl;

[0050] m is 1 or 2;

[0051] R 12 is selected from hydrogen or phenyl.

[0052] Furthermore, the structure of the compound is shown in Formula IVa:

[0053]

[0054] in,

[0055] R2 is selected from hydrogen, substituted or unsubstituted C1-C3 alkyl; the substituent of the alkyl is selected from -OC(O)R7, -OC(O)OR7;

[0056] R7 is selected from C1-C4 alkyl;

[0057] m is 1 or 2;

[0058] Preferably,

[0059] The structure of the compound is shown in Formula IVb:

[0060]

[0061] in,

[0062] m is 1 or 2.

[0063] Furthermore, the structure of the compound is shown in Formula V:

[0064]

[0065] in,

[0066] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C8 alkyl, C1-C8 alkoxy; n is an integer selected from 1 to 5;

[0067] R2 is selected from hydrogen, C1-C8 alkyl;

[0068] R3 and R4 are independently selected from hydrogen and C1-C8 alkyl;

[0069] R9 is selected from -C(S)NHR 10 , substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted 5-10 membered aryl, substituted or unsubstituted 3-10 membered cycloalkyl, substituted or unsubstituted 3-10 membered heterocycloalkyl; the alkyl substituent is selected from -C(S)NHR 10 、-C(O)OR 10 , 5-10 membered aryl, 5-10 membered heteroaryl; the substituents of the cycloalkyl and heterocycloalkyl groups are selected from hydroxyl, substituted or unsubstituted 5-10 membered aryl; the substituents of the aryl groups are selected from C1-C8 alkyl, halogen, hydroxyl, C1-C8 alkoxy, 5-10 membered aryl; the heteroatom of the heterocycloalkyl or heteroaryl group is N, O or S, and the number of the heteroatoms is 1, 2 or 3;

[0070] R 10 is selected from hydrogen, substituted or unsubstituted C1-C8 alkyl; the substituent of the alkyl is selected from -C(O)OR11 ;

[0071] R 11 Selected from C1 to C8 alkyl;

[0072] R2, R3, R4 are selected from hydrogen, R9 is selected from 3-membered cycloalkyl or When R1 and the benzene ring do not form

[0073] Preferably, the structure of the compound is as shown in Formula Va:

[0074]

[0075] in,

[0076] R1 is selected from halogen;

[0077] R2 is selected from hydrogen, C1-C3 alkyl;

[0078] R3 and R4 are independently selected from hydrogen and C1-C3 alkyl;

[0079] R9 is selected from -C(S)NHR 10 , naphthyl, tetrahydropyranyl, Substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted 3-6 membered cycloalkyl; the substituent of the alkyl is selected from -C(S)NHR 10 、-C(O)OR 10 , phenyl, The substituents of the 3-6 membered cycloalkyl group are selected from hydroxyl, substituted or unsubstituted phenyl; the substituents of the phenyl group are selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, and phenyl;

[0080] R 10 is selected from hydrogen, substituted or unsubstituted C1-C4 alkyl; the substituent of the alkyl is selected from -C(O)OR 11 ;

[0081] R 11 Selected from C1-C4 alkyl;

[0082] R2, R3, R4 are selected from hydrogen, R9 is selected from 3-membered cycloalkyl or When R1 and the benzene ring do not form

[0083] More preferably, the structure of the compound is as shown in Formula Vb:

[0084]

[0085] in,

[0086] R9 is selected from substituted C1-C4 alkyl groups; the substituent of the alkyl group is selected from phenyl groups.

[0087] Furthermore, the structure of the compound is shown in Formula VI:

[0088]

[0089] in,

[0090] R1 is a substituent at any position on the benzene ring, n is the number of substituents R1; each R1 is independently selected from halogen, C1-C8 alkyl, C1-C8 alkoxy; n is an integer selected from 1 to 5;

[0091] R2 is selected from hydrogen, substituted or unsubstituted C1-C8 alkyl; the substituent of the alkyl is selected from -OC(O)R7, -OC(O)OR7;

[0092] R7 is selected from hydrogen, C1-C8 alkyl;

[0093] R9 is selected from substituted C1-C8 alkyl groups; the substituent of the alkyl group is selected from 5-10 membered aryl groups;

[0094] Preferably, the structure of the compound is as shown in Formula VIa:

[0095]

[0096] in,

[0097] R1 is selected from halogen;

[0098] R9 is selected from substituted C1-C3 alkyl groups; the substituent of the alkyl group is selected from phenyl groups.

[0099] Furthermore, the compound is one of the following compounds:

[0100]

[0101]

[0102]

[0103] The present invention also provides use of the aforementioned compound, or its stereoisomer, or its tautomer, or its chiral isomer, or its salt in the preparation of an inhibitor of sirtuin 2-related protein;

[0104] Preferably, the inhibitor is a SIRT5 protein inhibitor.

[0105] The present invention also provides the use of the aforementioned compound, or its stereoisomer, or its tautomer, or its chiral isomer, or its salt in the preparation of a drug for preventing and / or treating cancer;

[0106] Preferably, the cancer is liver cancer, hepatoblastoma, breast cancer, lung cancer, pancreatic cancer, prostate cancer or leukemia.

[0107] The present invention also provides a drug, which is a preparation prepared with the aforementioned compound, or its stereoisomer, or its tautomer, or its chiral isomer, or its salt as an active ingredient, and pharmaceutically acceptable excipients or auxiliary ingredients.

[0108] The present invention also provides a pharmaceutical composition for preventing and / or treating cancer, comprising the aforementioned compound or its stereoisomers, tautomers or salts, or its prodrug molecules and a medically acceptable carrier.

[0109] The compounds and derivatives provided herein can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0110] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined herein, they should be given the meaning that a person skilled in the art would give them based on the disclosure and context.

[0111] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0112] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a ~C b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, C1-C4 alkyl refers to an alkyl group containing from 1 to 4 carbon atoms. In other words, C1-C4 alkyl includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0113] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of carbon atoms. For example, C1-C8 alkyl refers to an alkyl group having from 1 to 8 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. Alkyl groups can be straight or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl, among others.

[0114] "Halogen" is fluorine, chlorine, bromine or iodine.

[0115] "Cycloalkyl" refers to a saturated or partially saturated non-aromatic cyclic group consisting of carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused, bridged and spiro ring systems).

[0116] "Heterocycloalkyl" refers to a saturated or partially saturated non-aromatic cyclic group containing at least one heteroatom; including single rings or multiple rings (including fused, bridged, and spiro ring systems); wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom. Examples of heterocyclyl groups include, for example, piperidinyl, piperazinyl, and morpholinyl.

[0117] "Aryl" refers to a radical containing aromatic unsaturation having no ring heteroatoms and having a single ring or multiple rings (including fused, bridged and spiro ring systems), such as phenyl, anthracenyl, and naphthyl.

[0118] "Heteroaryl" refers to an aromatic unsaturated ring containing at least one heteroatom; including single rings or multiple rings (including fused, bridged, and spiro ring systems); wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom. Examples include pyridyl, pyrazinyl, pyridazinyl, pyrazolyl, furanyl, thienyl, and oxazolyl.

[0119] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that make up a pharmaceutical dosage form and physiologically compatible with the receptor.

[0120] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts of the above-mentioned compounds or their stereoisomers, formed with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final isolation and purification of the compound. They can also be obtained by mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent amounts). These salts may form a precipitate in the solution and be collected by filtration, or be recovered after evaporation of the solvent, or be obtained by freeze-drying after reaction in an aqueous medium. The salts described in the present invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluorides, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates or trifluoroacetates of the compound.

[0121] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include, but are not limited to, oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

[0122] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0123] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0124] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0125] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0126] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0127] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0128] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0129] The pharmaceutically acceptable excipients described in the present invention refer to substances contained in the dosage form in addition to the active ingredients.

[0130] The pharmaceutically acceptable auxiliary ingredients described herein have certain physiological activities, but their addition does not alter the dominant role of the pharmaceutical composition in treating a disease. Instead, they merely provide auxiliary benefits. These auxiliary benefits are simply a utilization of the known activity of the ingredients and are conventional adjuvant therapies in the medical field. The use of such auxiliary ingredients in conjunction with the pharmaceutical composition of the present invention remains within the scope of protection of the present invention.

[0131] Compared with the prior art, the present invention has the following beneficial effects:

[0132] The present invention provides a class of compounds with SIRT5 protein inhibitory activity, which can be used to prepare SIRT5 protein inhibitors and drugs for treating cancer. The present invention provides more options for the development and application of SIRT5 small molecule inhibitors and anticancer drugs.

[0133] Obviously, based on the above contents of the present invention, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0134] The following further describes the above content of the present invention in detail through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. DETAILED DESCRIPTION

[0135] The raw materials and equipment used in the specific embodiments of the present invention are all known products and are obtained by purchasing commercial products.

[0136] Example 1. Synthesis of Compound 1-2

[0137] Synthesis route: The substrate is the corresponding pyrimidine raw material

[0138]

[0139] 1. Synthesis of Compound 1

[0140] Methyl 2,4-dichloro-5-pyrimidinecarboxylate (1 eq), N-Boc-1,3-propylenediamine (1.2 eq), and triethylamine (1.5 eq) were dissolved in acetonitrile and allowed to react at room temperature for 30 min. The reaction was completed. The reaction solution was diluted with water and extracted with ethyl acetate (20 ml, 3 times). The organic layer was dried over anhydrous magnesium sulfate, concentrated, and purified by column chromatography (PE:EA = 6:1 → 4:1) to obtain a white solid (1b) in approximately 50% yield.

[0141] 1b (1 eq) was placed in a sealed tube and dissolved in an appropriate amount of methanol. Concentrated hydrochloric acid (15 μl / 200 mg) was added for activation at room temperature for 1 h. 2-Chloroaniline (1.5 eq) was then added, and the sealed tube was placed in a 115°C oil pan to react for 8 h. The reaction solution was then concentrated under reduced pressure, and 4 ml of DCM and 1 ml of TFA were added, stirring at room temperature for 30 min. The pH of the reaction solution was then adjusted to 8, and extracted with EA (20 ml, three times). The organic layers were combined, dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and purified by column chromatography (DCM:MeOH = 50:1 → 10:1) to obtain a white solid (1c). The overall yield for the two steps was approximately 62%.

[0142] 3-Alanine ethyl ester hydrochloride (1.2 eq) was dissolved in dry DCM and activated at room temperature for 40 min with triethylamine (3 eq) and carbon disulfide (1.5 eq). Triphosgene (0.36 eq) in DCM was then added under ice. After addition, activation was continued at room temperature for approximately 100 min. The reaction mixture was then concentrated to remove the solvent and residual triphosgene. A solution of 1c (1 eq) and triethylamine (3 eq) in DCM was then added, and the mixture was stirred overnight at room temperature. After completion of the reaction, the mixture was slowly purified by column chromatography (DCM:MeOH = 150:1) to afford 1d as a white solid in a 55% yield.

[0143] 1d (1 eq) and sodium hydroxide (2 eq) were dissolved in a 2:1 ethanol:water mixture and allowed to react at room temperature for 30 min. The pH of the reaction solution was adjusted to approximately 6 with dilute hydrochloric acid, and the ethanol in the reaction solution was removed as much as possible by vacuum concentration. The solution was then extracted with EA (20 ml, 3 times). The EA layer was then completely concentrated and purified by column chromatography (DCM:MeOH = 50:1) to obtain a white solid (71%), namely, compound 1. 1H NMR (400MHz, DMSO-d6) δ8.88(s,1H),8.53(s,1H),8.25(t,J=5.8Hz,1H),7.86( dd,J=8.1,1.5Hz,1H),7.63(s,1H),7.49(dd,J=8.0,1.5Hz,1H),7.45(t,J=5.7H z,1H),7.36(td,J=8.0,1.5Hz,1H),7.17(td,J=7.7,1.6Hz,1H),3.78(s,3H),3 .56(s,2H),3.39(q,J=6.7Hz,4H),2.47(t,J=6.7Hz,2H),1.77-1.69(m,2H)ppm. 13 C NMR (101MHz, DMSO-d6) δ173.81,166.78,161.60,161.47,160.78,136.32,129.79,127. 73,127.66,126.71,126.09,97.61,51.86,38.09,34.32,29.17,19.12,14.01ppm.HRMS m / z:calcd for C 19 H 23 ClN6O4S[M+H] + 467.1263found 467.1262.

[0144] 2. Synthesis of Compound 2

[0145] According to the synthesis method of compound 1, the raw material 2,4-dichloro-5-pyrimidinecarboxylic acid methyl ester was replaced with 2,4-dichloro-5-pyrimidinecarboxylic acid isopropyl ester to prepare compound 2.

[0146] Compound 2: 1 H NMR (400MHz, DMSO-d6) δ8.84 (s, 1H), 8.52 (s, 1H), 8.26 (t, J = 5.7Hz, 1H), 7.87 (dd, J=8.1,1.5Hz,1H),7.59(s,1H),7.49(dd,J=8.0,1.4Hz,1H),7.42(s,1H),7.35(td ,J=7.7,1.5Hz,1H),7.16(td,J=7.7,1.6Hz,1H),5.12–5.05(m,1H),3.56(s,2H),3 .39(q,J=6.3Hz,4H),2.46(s,2H),1.76–1.70m,2H),1.30(s,3H),1.29(s,3H)ppm. 13C NMR(101MHz,DMSO-d6)δ165.99,161.75,161.42,160.65,136.36,129.77,127.72 ,127.53,126.55,125.99,98.01,67.87,38.09,34.51,29.16,29.11,22.16.HRMS m / z:calcd forC 21 H 27 ClN6O4S[M+H] + 495.1976found 495.1978.

[0147] Example 2. Synthesis of Compound 4-17

[0148] The synthetic route is as follows: R is the corresponding aniline

[0149]

[0150] 1. Synthesis of compound 4

[0151] Ethyl 2,4-dichloro-5-pyrimidinecarboxylate (1 eq), N-Boc-1,3-propylenediamine (1.2 eq), and triethylamine (1.5 eq) were dissolved in acetonitrile and allowed to react at room temperature for 30 min. The reaction was completed. The reaction solution was diluted with water and extracted with ethyl acetate (20 ml, three times). The organic layer was dried over anhydrous magnesium sulfate, concentrated, and purified by column chromatography (PE:EA = 6:1 → 4:1) to obtain a white solid (4b) in approximately 57% yield.

[0152] 4b (1 eq) was placed in a sealed tube and dissolved in methanol. Concentrated hydrochloric acid (15 μl / 200 mg) was added for activation at room temperature for 1 h. 2-Fluoroaniline (1.5 eq) was then added, and the sealed tube was placed in a 115°C oil pan to react for 8 h. The reaction solution was then concentrated under reduced pressure, and 4 ml of DCM and 1 ml of TFA were added, stirring at room temperature for 30 min. The pH of the reaction solution was then adjusted to 8, and extracted with EA (20 ml, 3 times). The organic layers were combined, dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and purified by column chromatography (DCM:MeOH = 50:1 → 10:1) to obtain a white solid (4c). The overall yield for the two steps was approximately 77%.

[0153] 3-Alanine ethyl ester hydrochloride (1.2 eq) was dissolved in dry DCM and activated at room temperature for 40 min with triethylamine (3 eq) and carbon disulfide (1.5 eq). Triphosgene (0.36 eq) in DCM was then added under ice. After addition, activation was continued at room temperature for approximately 100 min. The reaction mixture was then concentrated to remove the solvent and residual triphosgene. A solution of 4c (1 eq) and triethylamine (3 eq) in DCM was then added, and the mixture was stirred overnight at room temperature. After completion of the reaction, the mixture was slowly purified by column chromatography (DCM:MeOH = 150:1) to afford 4d as a white solid in a 49% yield.

[0154] 4d (1 eq) and sodium hydroxide (2 eq) were dissolved in a 2:1 ethanol:water mixture and reacted at room temperature for 30 min. The pH of the reaction solution was adjusted to approximately 6 with dilute hydrochloric acid, and the ethanol in the reaction solution was removed as much as possible by vacuum concentration. The solution was then extracted with EA (20 ml, 3 times). The EA layer was then completely concentrated and purified by column chromatography (DCM:MeOH = 50:1) to obtain a white solid (66%), namely compound 4. 1 H NMR (400MHz, DMSO-d6) δ12.27(s,1H),9.21(s,1H),8.54(s,1H),8.25(t,J=5.8Hz,1H),7.85–7.76(m,1H),7.57(s,1H),7.40(d,J=5.7Hz,1H), 7.26–7.11(m,3H),4.24(q,J=7.1Hz,2H),3.57(s,2H),3.46–3.33(m,4H),2.48(d,J=6.6Hz,1H),1.78–1.68(m,2H),1.29(t,J=7.1Hz,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.66,166.44,161.69,161.48,160.68,156.69,154.25,127.42,127.31,126.15,125.65,1 25.58,124.51,124.47,115.99,115.79,97.56,60.40,55.38,41.60,38.04,34.11,29.07,14.67ppm.HRMSm / z:calcd for C 20 H 25 FN6O4S[M+H] + 465.1715found 465.1713.

[0155] 2. Synthesis of Compound 5-17

[0156] According to the synthesis method of compound 4, the raw material 2-fluoroaniline was replaced by 2-bromoaniline to prepare compound 5; the raw material 2-fluoroaniline was replaced by 2-methylaniline to prepare compound 6; the raw material 2-fluoroaniline was replaced by 3-fluoroaniline to prepare compound 7; the raw material 2-fluoroaniline was replaced by 3-chloroaniline to prepare compound 8; the raw material 2-fluoroaniline was replaced by 3-bromoaniline to prepare compound 9; the raw material 2-fluoroaniline was replaced by 3-methylaniline to prepare compound 10; the raw material 2-fluoroaniline was replaced by 3-methylaniline Compound 11 was prepared by replacing the raw material 2-fluoroaniline with 4-fluoroaniline; compound 12 was prepared by replacing the raw material 2-fluoroaniline with 4-chloroaniline; compound 13 was prepared by replacing the raw material 2-fluoroaniline with 4-methylaniline; compound 14 was prepared by replacing the raw material 2-fluoroaniline with 4-methoxyaniline; compound 15 was prepared by replacing the raw material 2-fluoroaniline with 2,6-dimethylaniline; compound 16 was prepared by replacing the raw material 2-fluoroaniline with 2,6-difluoroaniline.

[0157] Characterization data of compound 5: 1 H NMR (400MHz, DMSO-d6) δ8.82(s,1H),8.52(s,1H),8.25(t,J=5.7Hz,1H),7.89–7 .69(m,2H),7.65(dd,J=8.0,1.5Hz,1H),7.54(t,J=5.6Hz,1H),7.40(td,J=7.7,1 .5Hz,1H),7.10(td,J=7.7,1.6Hz,1H),4.24(q,J=7.1Hz,2H),3.54(s,2H),3.43– 3.32(m,4H),2.42(t,J=6.6Hz,2H),1.76–1.68(m,2H),1.29(t,J=7.1Hz,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ174.17,166.40,161.66,161.47,160.73,137.67,132.96,128 .38,127.03,126.60,118.81,97.69,60.43,38.06,34.84,29.11,26.81,14.69ppm.HRMS m / z:calcd for C 20 H 25 BrN6O4S[M+H] + 525.0914found 525.0915and 527.0896.

[0158] Characterization data of compound 6: 11H NMR (400 MHz, DMSO-d6) δ 12.22 (br s, 1H), 8.94 (s, 1H), 8.51 (s, 1H), 8.19 (t, J = 5.7 Hz, 1H), 7.68–7.50 (m, 2H), 7.42 (d, J = 5.8 Hz, 1H), 7.23–7.14 (m, 2H), 7.06 (td, J = 7.4, 1.3 Hz, 1H), 4.23 (q, J = 7.1 Hz, 2H), 3.62–3.52 (m, 2H), 3.42–3.30 (m, 4H), 2.48 (t, J = 6.7 Hz, 2H), 2.23 (s, 3H), 1.79–1.65 (m, 2H), 1.28 (t, J = 7.1 Hz, 3H) ppm. 13 13C NMR (101 MHz, DMSO-d6) δ 173.71, 166.54, 161.94, 161.77, 160.74, 137.74, 132.60, 130.59, 126.19, 125.96, 125.12, 96.88, 60.26, 37.85, 34.15, 29.21, 18.50, 14.71 ppm. HRMS m / z: calcd for C 21 H 28 N6O4S [M + H] + 461.1966 found 461.1959.

[0159] Characterization data of compound 7: 1 1H NMR (400 MHz, DMSO-d6) δ 9.97 (s, 1H), 8.59 (s, 1H), 8.37 (t, J = 5.7 Hz, 1H), 7.81 (dt, J = 12.4, 2.4 Hz, 1H), 7.65 (d, J = 5.0 Hz, 1H), 7.54 (dd, J = 8.3, 1.9 Hz, 1H), 7.44 (t, J = 5.5 Hz, 1H), 7.32 (q, J = 8.0 Hz, 1H), 6.79 (td, J = 8.4, 2.6 Hz, 1H), 4.26 (q, J = 7.1 Hz, 2H), 3.62–3.50 (m, 6H), 2.47 (t, J = 6.7 Hz, 2H), 1.89–1.78 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H) ppm. 13C NMR(101MHz,DMSO-d6)δ173.76,166.35,163.87,161.67,161.48,160.84,160.45,142.38,142.26,130.52,130 .43,115.55,108.78,108.56,106.49,106.22,97.65,60.50,55.37,41.76,38.45,34.19,29.07,14.67ppm.HRMS m / z:calcd forC 20 H 21 FN6O4S[M+H] + 465.1712found 465.1708.

[0160] Characterization data of compound 8: 1 H NMR (400MHz, DMSO-d6) δ12.25(s,1H),9.97(s,1H),8.59(s,1H),8.38(t,J=5. 8Hz,1H),8.08(t,J=2.0Hz,1H),7.73–7.54(m,2H),7.42(t,J=5.6Hz,1H),7.32 (t,J=8.1Hz,1H),7.02(dd,J=8.0,2.0Hz,1H),4.25(q,J=7.1Hz,2H),3.66–3. 43(m,6H),2.48(t,J=6.7Hz,2H),1.92–1.77(m,2H),1.30(t,J=7.0Hz,3H)ppm. 13 C NMR (101MHz, DMSO-d6) δ173.65,166.33,161.65,160.80,160.46,142.02,133.44,130.56,121. 91,119.09,118.11,97.60,60.50,41.84,38.46,34.10,29.27,29.14,14.67ppm.HRMSm / z:calcd for C 20 H 25 ClN6O4S[M+H] + 481.1419found 481.1416.

[0161] Characterization data of compound 9: 1H NMR (400MHz, DMSO-d6) δ12.27(s,1H),9.96(s,1H),8.59(s,1H),8.38(t,J=5.8 Hz,1H),8.26(t,J=2.0Hz,1H),7.72–7.63(m,2H),7.48(t,J=5.5Hz,1H),7.26(t ,J=8.1Hz,1H),7.17–7.12(m,1H),4.26(q,J=7.1Hz,2H),3.53(dt,J=14.3,7.1 Hz, 6H), 2.48 (d, J = 6.7Hz, 1H), 1.84 (p, J = 6.8Hz, 2H), 1.30 (t, J = 7.1Hz, 3H) ppm. 13 CNMR(101MHz,DMSO-d6)δ173.62,166.33,161.63,160.77,160.47,142.16,130.87,124.79,121. 99,121.96,118.49,97.54,60.50,55.35,38.45,34.12,29.12,26.81,14.67ppm.HRMSm / z:calcd for C 20 H 25 BrN6O4S[M+H] + 525.0914found 525.0913and 527.0896.

[0162] Characterization data of compound 10: 1 H NMR(400MHz,DMSO-d6)δ12.26(br s,1H),9.70(s,1H),8.57(s,1H),8.32(t,J=5.7Hz,1H),7.69(s,1H),7. 61(s,1H),7.57–7.53(m,1H),7.41(t,J=5.7Hz,1H),7.18(t,J=7.8Hz,1H ),6.83–6.79(m,1H),4.25(q,J=7.1Hz,2H),3.60–3.44(m,6H),2.47(t, J=6.7Hz,2H),2.29(s,3H),1.90–1.79(m,2H),1.30(t,J=7.1Hz,3H)ppm. 13C NMR(101MHz,DMSO-d6)δ173.87,166.44,161.75,160.96,160.42,140.31,138.00,128.79,1 23.25,120.46,117.17,96.92,60.36,55.37,41.79,38.38,34.19,29.17,21.82,14.69.HRMS m / z:calcd for C 21 H 28 N6O4S[M+H] + 461.1966found 461.1967.

[0163] Characterization data of compound 11: 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),9.79(s,1H),8.57(s,1H),8.37(t,J=5.9Hz,1H),7. 62(s,1H),7.54(t,J=2.2Hz,1H),7.44(t,J=5.6Hz,1H),7.33(ddd,J=8.2,2.4,0.9Hz,1H), 7.20(t,J=8.1Hz,1H),6.58(ddd,J=8.2,2.5,0.9Hz,1H),4.25(q,J=7.1Hz,2H),3.74(s,3 H),3.66–3.48(m,6H),2.48(t,J=6.8Hz,1H),1.88–1.78(m,2H),1.30(t,J=7.1Hz,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.62,166.35,161.68,160.57,159.96,141.47,129.72,112.3 9,108.08,105.73,97.15,60.47,55.42,41.55,38.45,34.09,29.05,14.68,14.54.HRMS m / z:calcd for C 21 H 28 N6O5S[M+H] + 477.1915found477.1916.

[0164] Characterization data of compound 12: 11H NMR (400 MHz, DMSO-d6) δ 12.27 (s, 1H), 9.84 (s, 1H), 8.56 (s, 1H), 8.33 (d, J = 5.7 Hz, 1H), 7.82–7.77 (m, 2H), 7.73 (t, J = 5.5 Hz, 1H), 7.54 (d, J = 6.6 Hz, 1H), 7.15 (t, J = 8.9 Hz, 2H), 4.25 (q, J = 7.1 Hz, 2H), 3.57 (s, 2H), 3.54–3.45 (m, 4H), 2.48 (t, J = 6.8 Hz, 2H), 1.85–1.76 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H) ppm. 13 13C NMR (101 MHz, DMSO-d6) δ 173.55, 166.39, 161.70, 160.70, 160.26, 159.16, 156.79, 136.72, 121.64, 115.58, 115.36, 97.18, 60.42, 41.72, 38.43, 34.14, 29.21, 14.67 ppm. HRMS m / z: calcd for C 20 H 25 FN6O4S [M + H] + 465.1715 found 465.1714.

[0165] Characterization data of compound 13: 1 1H NMR (400 MHz, DMSO-d6) δ 9.90 (s, 1H), 8.57 (s, 1H), 8.32 (t, J = 5.9 Hz, 1H), 7.85–7.80 (m, 2H), 7.69 (d, J = 5.7 Hz, 1H), 7.51 (t, J = 5.7 Hz, 1H), 7.38–7.33 (m, 2H), 4.24 (q, J = 7.1 Hz, 2H), 3.57 (s, 2H), 3.54–3.45 (m, 4H), 2.48 (t, J = 6.8 Hz, 2H), 1.86–1.77 (m, 2H), 1.29 (t, J = 7.1 Hz, 3H) ppm. 13 13C NMR (101 MHz, DMSO-d6) δ 182.49, 173.68, 166.38, 161.67, 160.81, 160.44, ; 139.43, 128.82, 126.01, 121.31, 97.34, 60.45, 41.64, 38.48, 34.19, 29.20, 14.67 ppm. HRMS m / z: calcd for C 20 H 25 ClN6O4S [M + H]+ 481.1419found481.1414.

[0166] Characterization data of compound 14: 1 H NMR (400MHz, DMSO-d6) δ12.23(s,1H),9.67(s,1H),8.55(s,1H),8.30(d,J=5. 8Hz,1H),7.67(d,J=8.1Hz,2H),7.62(s,1H),7.43(t,J=5.8Hz,1H),7.11(d,J= 8.2Hz,2H),4.24(q,J=7.1Hz,2H),3.57(s,2H),3.54–3.44(q,J=6.6Hz,4H),2. 48(t,J=6.6Hz,1H),2.25(s,3H),1.88–1.76(m,2H),1.30(t,J=7.1Hz,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.65,166.47,161.75,160.94,160.43,137.83,131.39, 129.38,120.01,96.63,60.33,41.77,38.38,34.12,29.10,20.86,14.70ppm.HRMS m / z:calcd forC 21 H 28 N6O4S[M+H] + 461.1966found 461.1961.

[0167] Characterization data of compound 15: 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),9.64(s,1H),8.54(s,1H),8.30(s,1H),7.68(d,J=8.5Hz,2H),7.61(s,1H),7.42(s,1H),6.93–6.88(m, 2H),4.24(q,J=7.1Hz,2H),3.73(s,3H),3.57(s,2H),3.53–3.43(m,4H),2.48(t,J=6.8Hz,1H),1.86–1.77(m,2H),1.30(t,J=7.1Hz,3H)ppm. 13C NMR(101MHz,DMSO-d6)δ173.63,166.45,161.74,160.72,160.25,155.11,133.40, 121.45,114.18,60.31,56.50,55.64,38.37,34.09,29.17,19.03,14.70ppm.HRMS m / z:calcd for C 21 H 28 N6O5S[M+H] + 477.1915found477.1910.

[0168] Characterization data of compound 16: 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.86(d,J=58.9Hz,1H),8.45(d,J=61 .0Hz,1H),8.11(d,J=31.3Hz,1H),7.68–7.31(m,2H),7.07(s,3H),4.22(q,J =7.1Hz,2H),3.73–3.47(m,4H),3.19–3.01(m,2H),2.48(d,J=5.3Hz,1H),2. 13(s,6H),1.88–1.75(m,1H),1.55–1.42(m,1H),1.26(q,J=8.51Hz,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.74,166.65,162.61,161.90,160.97,136.68,136.44,136 .01,128.02,126.63,95.94,60.14,55.37,37.61,34.18,29.03,18.74,14.73ppm.HRMS m / z:calcd forC 22 H 30 N6O4S[M+H] + 475.2122found 475.2124.

[0169] Characterization data of compound 17: 1 H NMR(400MHz,DMSO-d6)δ9.46–9.01(m,1H),8.59–8.41(m,1H),8.27–8.13(m,1H),7.80–6.76(m,6H) ,4.31–4.16(m,2H),3.85–3.38(m,6H),2.50–2.40(m,2H),1.77–1.54(m,2H),1.33–1.20(m,3H)ppm.13 C NMR (101MHz, DMSO-d6) δ173.68,166.46,162.10,161.68,160.85,160.13,157.67,127.87,116. 59,116.43,116.27,112.23,112.18,112.00,97.60,60.39,37.87,34.14,28.97,14.66ppm.HRMS m / z:calcd forC 20 H 24 F2N6O4S[M+H] + 483.1621found 483.1619.

[0170] Example 3. Synthesis of Compounds 18-20

[0171] The synthetic route is as follows: R is the corresponding aniline

[0172]

[0173] 1. Synthesis of Compound 18

[0174] 3-(3-(5-(ethoxycarbonyl)-2-(3-methylanilino)pyrimidin-4-ylamino)propyl)thioureido)propanoic acid (i.e., compound 10, 1 eq) and sodium hydroxide (2 eq) were dissolved in a 2:1 ethanol:water mixture and reacted at 80°C for 30 min. The pH of the reaction solution was adjusted to approximately 6 with dilute hydrochloric acid, resulting in the precipitation of a large amount of white solid. The filter cake was filtered, washed with ethyl acetate and dichloromethane, and dried to yield 18 as a white solid in 82% yield. 1 H NMR (400MHz, DMSO-d6) δ10.61(s,1H),9.07(s,1H),8.58(s,1H),7.73(t,J=5.4Hz,1H),7.58–7.52(m,2H),7.48(dd,J=8.0,2.1Hz,1H),7.26(t,J =7.8Hz,1H),6.93(dd,J=7.4,1.5Hz,1H),3.55(p,J=7.2,6.4Hz,4H),3.45(s,2H),2.47(d,J=6.8Hz,2H),2.31(s,3H),1.83(p,J=6.8Hz,2H)ppm. 13C NMR(101MHz,DMSO-d6)δ173.56,167.14,161.25,155.17,153.52,138.55,138 .32,129.17,124.95,121.40,118.11,98.79,38.95,34.11,28.90,21.69ppm.

[0175] 2. Synthesis of Compounds 19 and 20

[0176] Referring to the synthesis method of compound 18, compound 10 was replaced by 3-(3-(5-(ethoxycarbonyl)-2-(3-methoxyanilino)pyrimidin-4-yl-amino)propyl)thioureido)propionic acid 3-(3-(5-(ethoxycarbonyl)-2-(3-methylanilino)pyrimidin-4-yl-amino)propyl)thioureido)propionic acid (11) to prepare compound 19; compound 10 was replaced by 3-(3-(5-(ethoxycarbonyl)-2-(2-chloroanilino)pyrimidin-4-yl-amino)propyl)thioureido)propionic acid 3-(3-(5-(ethoxycarbonyl)-2-(3-methylanilino)pyrimidin-4-yl-amino)propyl)thioureido)propionic acid to prepare compound 20.

[0177] Characterization data of compound 19: 1 H NMR (400MHz, DMSO-d6) δ9.47(s,1H),9.16(d,J=5.9Hz,1H),8.50(s,1H),8.03(s,1H),7.84(s,1H),7.59(t,J=2.3Hz,1H),7.32(dd,J=8.1,1 .9Hz,1H),7.17(t,J=8.2Hz,1H),6.51(dd,J=8.2,2.5Hz,1H),3.74(s,3H),3.59–3.44(m,6H),2.48(t,J=6.8Hz,2H),1.86–1.79(m,2H)ppm. 13 C NMR (101MHz, DMSO-d6) δ173.97,169.84,162.41,160.46,159.95,159.62,142.23,129.61,111.90,107.34,105.12,55.36,38.25,34.49ppm.

[0178] Characterization data of compound 20: 1H NMR(400MHz,DMSO-d6)δ12.41(br s,1H),9.19(s,1H),8.66(t,J=5.8Hz,1H),8.53(s,1H),7.88(dd,J=8.1,1 .6Hz,1H),7.61(q,J=11.2,8.2Hz,1H),7.52(dd,J=8.0,1.4Hz,1H),7.47(t ,J=5.6Hz,1H),7.38(td,J=7.8,1.5Hz,1H),7.20(td,J=7.8,1.6Hz,1H),3. 56(s,2H),3.44–3.31(m,4H),2.47(t,J=6.7Hz,2H),1.76–1.69(m,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.60,167.87,161.71,159.15,157.90,135.65,129.90,127.89,127.63,126.73,126.52,98.75,38.28,34.11,29.06.HRMS m / z:calcd forC 18 H 21 ClN6O4S[M+H] + 453.1106found 453.1108.

[0179] Example 4. Synthesis of Compounds 21 and 22

[0180] The synthesis method is as follows: R is derived from the corresponding raw material amine

[0181]

[0182] 1. Synthesis of Compound 21

[0183] Intermediate 7c (1 eq) and 4-dimethylaminopyridine (0.1 eq) were dissolved in dry dichloromethane. Triethylamine (2 eq) and di-tert-butyl dicarbonate (1.1 eq) were added with stirring and allowed to react at room temperature for 2 h. The reaction solution was concentrated and purified by column chromatography (PE:EA = 4:1 → 2:1) to afford white intermediate 7e in 91% yield.

[0184] 7e (1 eq) and sodium hydroxide (2 eq) were dissolved in a 2:1 ethanol:water mixture and refluxed at 80°C for 3 h. The reaction mixture was concentrated under reduced pressure to remove as much ethanol as possible. The pH was adjusted to 6 with dilute hydrochloric acid, filtered, and the filter cake was washed with ethyl acetate and dichloromethane. The filter cake was dried to obtain the white intermediate 7f in 88% yield.

[0185] 7f (1 eq), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (1.2 eq), and triethylamine (3 eq) were dissolved in an appropriate amount of dichloromethane and activated at room temperature for 20 min. Cyclopropylamine was added with stirring, and the reaction was continued at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by column chromatography (PE:EA = 4:1 → 1:1) to obtain the white intermediate 21a in 87% yield.

[0186] 21a was dissolved in dichloromethane (4 ml), and TFA (1 ml) was added dropwise with stirring. The reaction was allowed to proceed at RT for 30 min. The reaction was completed by monitoring the complete conversion of the starting material by TLC. The pH value of the reaction solution was adjusted to 8 with saturated sodium bicarbonate, and an appropriate amount of saturated brine was added. The solution was extracted with EA (20 ml, 3 times). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the white intermediate 21b (90%).

[0187] The subsequent steps refer to the synthesis method of compound 4, replacing intermediate 4c with 21b to prepare thiourea intermediate 21c, and replacing intermediate 4d with 21c to prepare target compound 21. 1 H NMR (400MHz, DMSO-d6) δ9.69(s,1H),9.06(t,J=4.2Hz,1H),8.47(s,1H),8.27(d,J=4.0Hz,1H),7.8 3(dt,J=12.4Hz,J=2.0Hz,1H),7.65(t,J=5.6Hz,1H),7.54(dd,J=8.0Hz,J=1.2Hz,1H),7.42(t,J=5. 6Hz,1H),7.30(q,J=8.0Hz,1H),6.74(td,J=8.4Hz,J=2.0Hz,1H),3.57(s,2H),3.52-3.44(m,4H),2. 81-2.74(m,1H),2.48(t,J=6.4Hz,2H),1.88-1.78(m,2H),0.72-0.64(m,2H),0.58-0.52(m,2H)ppm. 13 C NMR (101MHz, DMSO-d6) δ173.76,168.40,163.92,161.62,161.54,160.14,156.72,142.88,142.77,130.42,130. 32,115.12,108.14,107.93,106.03,105.76,101.07,60.30,46.02,38.30,34.28,29.08,23.08,14.54,6.17ppm.

[0188] 2. Synthesis of Compound 22

[0189] Referring to the synthesis method of compound 21, the raw material cyclopropylamine was replaced with benzylamine to obtain the target compound 22. 1 H NMR (400MHz, DMSO-d6) δ9.72(s,1H),9.05(t,J=4.2Hz,1H),8.87(t,J=6.0Hz,1H),8.60(s,1 H),7.84(dt,J=12.4Hz,J=2.0Hz,1H),7.71(s,1H),7.54(dd,J=8.0Hz,J=1.6Hz,1H),7.45(t, J=5.6Hz,1H),7.36-7.28(m,5H),7.28-7.22(m,1H),6.75(td,J=8.4Hz,J=1.2Hz,1H),4.45(d ,J=6.0Hz,2H),3.56(s,2H),3.52-3.43(m,4H),2.45(t,J=5.6Hz,2H),1.89-1.75(m,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.92,167.06,163.93,161.77,161.54,160.21,156.77,142.87,142.75,140.12,130.44,130 .34,128.75,127.71,127.20,115.18,108.19,107.98,106.08,105.81,101.04,42.62,38.33,34.50,34.44,29.09ppm.

[0190] Example 5. Synthesis of Compounds 23-39, 41-49, 51, 53-59

[0191] The synthesis method is as follows: R1 is derived from the corresponding amine raw material, only R2 and R3 of compound 59 are methyl, and R2 and R3 of compounds 23-39, 41-49, 51, 53-58 are H

[0192]

[0193] 1. Synthesis of Compound 23

[0194] Referring to the synthesis method of compound 4, 2-fluoroaniline was replaced with 2-chloroaniline to prepare intermediate 23c. Referring to the synthesis method of compound 21, intermediate 7c was replaced with 23c, and the raw material cyclopropylamine in subsequent steps was replaced with n-propylamine to prepare compound 23 by the same method. 1H NMR (400MHz, DMSO-d6) δ12.26(s,1H),9.01(t,J=5.7Hz,1H),8.47(s,2H),8.32(t,J=5.6Hz ,1H),8.01(dd,J=8.2,1.6Hz,1H),7.64(t,J=5.3Hz,1H),7.53–7.42(m,2H),7.35(td,J=7.8 ,1.5Hz,1H),7.11(td,J=7.7,1.6Hz,1H),3.57(s,2H),3.40-3.34(m,4H),3.16(q,J=6.6Hz ,2H),2.48(t,J=6.8Hz,2H),1.77–1.70m,2H),1.56-1.47(m,2H),0.88(t,J=7.4Hz,3H)ppm. 13 C NMR (101MHz, DMSO-d6) δ184.26,173.59,166.96,161.76,160.27,156.46,136.65,129.68,127. 75,126.15,125.22,125.08,101.63,41.85,40.99,38.03,34.14,29.15,22.84,11.93ppm.HRMS m / z:calcd for C 21 H 28 ClN7O3S[M+H] + 494.1736found 494.1734.

[0195] 2. Synthesis of compounds 24-39, 41-49, 51, 53-59

[0196] Referring to the synthetic method of compound 23, the raw material n-propylamine was replaced by isobutylamine to prepare compound 24; the raw material n-propylamine was replaced by cyclobutylamine to prepare compound 25; the raw material n-propylamine was replaced by pyrrolidine to prepare compound 26; the raw material n-propylamine was replaced by (1S, 2S)-2-hydroxycyclohexylamine to prepare compound 27; the raw material n-propylamine was replaced by tetrahydropyran to prepare compound 28; the raw material n-propylamine was replaced by aniline to prepare compound 29; the raw material n-propylamine was replaced by 2-methylaniline to prepare compound 30; the raw material n-propylamine was replaced by 2-methoxyaniline to prepare compound 31; the raw material n-propylamine was replaced by 2-isopropylaniline to prepare compound 32; the raw material The raw material n-propylamine was replaced by 2-naphthylamine to prepare compound 33; the raw material n-propylamine was replaced by 1-naphthylamine to prepare compound 34; the raw material n-propylamine was replaced by 2-benzidine to prepare compound 35; the raw material n-propylamine was replaced by 3,5-dimethoxyaniline to prepare compound 36; the raw material n-propylamine was replaced by (S)-α-methylbenzylamine to prepare compound 37; the raw material n-propylamine was replaced by (R)α-methylbenzylamine to prepare compound 38; the raw material n-propylamine was replaced by (S)-α-ethylbenzylamine to prepare compound 39; the raw material n-propylamine was replaced by (R)-α-ethylbenzylamine to prepare compound 41; the raw material n-propylamine was replaced by 1-phenyl-3-butylamine to prepare compound 4 2; the raw material n-propylamine was replaced by L-phenylalanine hydrochloride to prepare compound 43; the raw material n-propylamine was replaced by (S)-2-tert-butoxycarbonyl-1-phenylethylamine hydrochloride to prepare compound 44; the raw material n-propylamine was replaced by (R)-2-tert-butoxycarbonyl-1-phenylethylamine hydrochloride to prepare compound 45; the raw material n-propylamine was replaced by 1-phenylcyclopropylamine to prepare compound 46; the raw material n-propylamine was replaced by (1R, 2S) 2-phenylcyclopropylamine to prepare compound 47; the raw material n-propylamine was replaced by (1R, 2S)-2-(3,4-difluorophenyl)cyclopropylamine to prepare compound 48; the raw material n-propylamine was replaced by 2-phenylpyrrolidine to prepare compound 49; the raw material The raw material n-propylamine was replaced by 3-phenylpyrrolidine to prepare compound 51; the raw material n-propylamine was replaced by (R)-3-phenylpyrrolidine to prepare compound 53; the raw material n-propylamine was replaced by 3-phenylpiperidine to prepare compound 54; the raw material n-propylamine was replaced by 4-phenylpiperidine to prepare compound 55; the raw material n-propylamine was replaced by 2-(1H-indol-2-yl)ethyl-1-amine to prepare compound 56; the raw material n-propylamine was replaced by (1R,2S)-1-amino-2,3-dihydro-1H-inden-2-ol to prepare compound 57; the raw material n-propylamine was replaced by (R)-2-amino-3-(1H-indol-3-yl)propionic acid methyl ester hydrochloride to prepare compound 58.Referring to the synthesis method of compound 41, compound 59 was prepared by replacing the raw material 3-alanine ethyl ester hydrochloride with 2,2-dimethyl-3-alanine ethyl ester hydrochloride.

[0197] Characterization data of compound 24: 1 H NMR(400MHz,DMSO-d6)δ12.24(br s,1H),8.97(t,J=5.5Hz,1H),8.52–8.43(m,2H),8.30(t,J=5.7Hz,1H),8.01 (d,J=7.9Hz,1H),7.58(s,1H),7.48(d,J=7.9Hz,1H),7.40(t,J=5.7Hz,1H), 7.37–7.32(m,1H),7.13–7.06(m,1H),3.57(s,2H),3.40–3.33(m,4H),3.06– 3.00(m,2H),2.51–2.47(m,3H),1.87–1.71(m,4H),0.88(d,J=6.4Hz,6H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.81,167.04,161.76,160.42,156.65,136.71,129.70,127.73,12 6.28,125.37,125.12,101.57,55.37,46.70,41.89,37.99,34.20,29.12,28.55,20.71.HRMS m / z:calcd for C 22 H 30 ClN7O3S[M+H] + 508.1892found508.1891.

[0198] Characterization data of compound 25: 1 H NMR(400MHz,DMSO-d6)δ12.25(br s,1H),9.92(s,1H),9.73(s,1H),8.93(d,J=7.2Hz,1H),8.61(s,1H),7. 89–7.85(m,1H),7.64–7.56(m,2H),7.49–7.42(m,2H),7.31(td,J=7.9,1 .4Hz,1H),4.39–4.29(m,1H),3.55(s,2H),3.40–3.33(m,4H),2.47(t,J= 6.8Hz,2H),2.25–2.19(m,2H),2.11–2.02(m,2H),1.75–1.66(m,4H)ppm. 13CNMR(101MHz,DMSO-d6)δ173.51,163.69,160.91,152.63,145.16,133.79,130.21,12 8.26,127.93,127.05,102.36,44.93,39.02,34.14,30.22,28.65,15.33,14.55.HRMS m / z:calcdfor C 22 H 28 ClN7O3S[M+H] + 506.1736found 506.1715.

[0199] Characterization data of compound 26: 1 H NMR (400MHz, DMSO-d6) δ8.33 (s, 1H), 8.11 (s, 1H), 8.05 (dd, J = 8.2, 1.6Hz, 1H), 7.74 (t, J=5.7Hz,1H),7.58(s,1H),7.47(dd,J=8.0,1.5Hz,1H),7.42(t,J=5.6Hz,1H),7.34(td ,J=7.9,1.5Hz,1H),7.09(td,J=7.7,1.6Hz,1H),3.56(s,2H),3.49(t,J=6.4Hz,4H),3. 35(q,J=6.5Hz,4H),2.47(t,J=6.7Hz,2H),1.84(t,J=6.3Hz,4H),1.77-1.70(m,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.78,166.60,160.77,159.71,156.22,136.78,129.67,127.74 ,125.93,125.10,124.88,104.96,55.38,47.69,41.89,38.13,34.19,29.15,25.77.HRMS m / z:calcd for C 22 H 28 ClN7O3S[M+H] + 506.1736found506.1735.

[0200] Characterization data of compound 27: 11H NMR (400 MHz, DMSO-d6) δ 8.98 (t, J = 5.7 Hz, 1H), 8.52 (s, 1H), 8.43 (s, 1H), 8.03 (dd, J = 8.2, 1.6 Hz, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.62 (s, 1H), 7.47 (dd, J = 8.0, 1.5 Hz, 1H), 7.43 (t, J = 5.6 Hz, 1H), 7.35 (td, J = 8.0, 1.5 Hz, 1H), 7.11 (td, J = 7.8, 1.6 Hz, 1H), 4.65 (s, 1H), 3.58 (t, J = 9.5 Hz, 3H), 3.41 - 3.34 (m, 5H), 2.47 (t, J = 6.8 Hz, 2H), 1.92–1.90 (m, 1H), 1.85–1.79 (m, 1H), 1.77–1.70 (m, 2H), 1.66–1.62 (m, 2H), 1.27–1.20 (m, 4H) ppm. 13 13C NMR (101 MHz, DMSO-d6) δ 173.68, 166.84, 161.75, 160.35, 156.93, 136.76, 129.70, 127.74, 126.10, 125.15, 125.01, 101.88, 71.47, 55.24, 46.11, 38.00, 34.94, 34.17, 31.81, 29.17, 24.98, 24.63, 9.17 ppm. HRMS m / z: calcd for C 24 H 32 ClN7O4S [M + H] + 550.1998 found 550.2000.

[0201] Characterization data of compound 28: 1 1H NMR (400 MHz, DMSO-d6) δ 8.93 (t, J = 5.6 Hz, 1H), 8.49 (s, 1H), 8.44 (s, 1H), 8.10 (d, J = 7.5 Hz, 1H), 8.02 (dd, J = 8.2, 1.6 Hz, 1H), 7.69 (d, J = 5.4 Hz, 1H), 7.59–7.42 (m, 2H), 7.34 (td, J = 7.8, 1.6 Hz, 1H), 7.11 (td, J = 7.7, 1.6 Hz, 1H), 3.99–2.92 (m, 2H), 3.90–3.85 (m, 3H), 3.56 (s, 2H), 3.40–3.36 (m, 4H), 2.47 (t, J = 6.7 Hz, 2H), 1.79–1.71 (m, 4H), 1.61–1.50 (m, 2H) ppm. 13C NMR (101MHz, DMSO) δ182.59,173.79,166.44,161.78,160.45,156.91,136.70,129.69,127.7 3,126.22,125.28,125.10,101.46,66.64,45.80,43.32,41.96,38.02,34.45,32.92,29.14.

[0202] Characterization data of compound 29: 1 H NMR(400MHz,DMSO-d6)δ10.00(s,1H),8.71(t,J=5.7Hz,1H),8.66(s,1H),8 .62(s,1H),7.98(dd,J=8.1,1.6Hz,1H),7.71–7.64(m,3H),7.52–7.45(m,2 H),7.39–7.31(m,3H),7.14(td,J=7.7,1.6Hz,1H),7.08(t,J=7.4Hz,1H),3 .56(s,2H),3.42–3.37(m,4H),2.46(t,J=6.7Hz,2H),1.79–1.72(m,2H)ppm. 13 C NMR(101MHz,DMSO-d6)δ173.75,166.08,161.80,160.68,157.85,139.40,136.64,129.73,128.9 5,127.76,126.79,125.87,125.46,123.95,121.24,101.72,41.79,38.12,34.44,29.18ppm.HRMS m / z:calcd for C 24 H 26 ClN7O3S[M+H] + 528.1579found 528.1565.

[0203] Characterization data of compound 30: 1H NMR (400MHz, DMSO-d6) δ9.66(s,1H),8.80(t,J=5.7Hz,1H),8.68(s,1H),8.59(s,1H),7.98(dd,J=8.1,1.6Hz,1H),7.70–7.56(m,1H),7.50(dd,J=8.0 ,1.5Hz,1H),7.42–7.34(m,2H),7.28(m,2H),7.17(m,3H),3.54(s,2H),3. 38(q,J=6.3Hz,4H),2.48(t,J=7.2Hz,2H),2.22(s,3H),1.77–1.70(m,2H). 13 C NMR (101MHz, DMSO) δ166.10,161.91,160.72,157.63,136.66,136.59,134.39,130.73,129.74,1 27.77,127.31,126.75,126.41,125.81,125.43,101.39,55.36,38.07,34.40,29.15,18.39.HRMS m / z:calcd for C 25 H 28 ClN7O3S[M+H] + 542.1736found 542.1712.

[0204] Characterization data of compound 31: 1 H NMR(400MHz,DMSO-d6)δ12.25(s,1H),9.38(s,1H),8.83(s,1H),8.72(s,1H),8.64(s,1H ),7.96(dd,J=8.2,1.6Hz,1H),7.58(dd,J=7.8,1.7Hz,2H),7.51(dd,J=8.0,1.5Hz,1H), 7.44–7.33(m,2H),7.18(m,2H),7.08(dd,J=8.4,1.4Hz,1H),6.95(td,J=7.6,1.4Hz,1H) ,3.82(s,3H),3.55(s,2H),3.41–3.36(m,4H),2.48(t,J=6.7Hz,2H),1.77–1.70(m,2H). 13C NMR (101MHz, DMSO-d6) δ173.62,165.79,161.74,152.65,129.80,127.80,126. 83,126.47,126.11,125.93,120.55,111.94,101.58,56.11,38.11,34.08.HRMS m / z:calcd for C 25 H 28 ClN7O4S[M+H] + 558.1685found 558.1692.

[0205] Characterization data of compound 32: 1 H NMR (400MHz, DMSO-d6) δ12.21(s,1H),9.70(s,1H),8.84(t,J=5.7Hz,1H),8.70(s,1H),8.58(s,1H), 7.99(dd,J=8.2,1.6Hz,1H),7.60(t,J=5.2Hz,1H),7.50(dd,J=8.0,1.5Hz,1H),7.42(t,J=5.6Hz,1H ),7.38–7.34(m,2H),7.29–7.25(m,1H),7.22–7.19(m,2H),7.14(td,J=7.7,1.6Hz,1H),3.56(s,2H) ,3.38(q,J=6.5Hz,4H),3.14(m,1H),2.48(t,J=7.3Hz,2H),1.77–1.71(m,2H),1.16(d,J=6.9Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ173.67,166.79,161.94,160.74,157.52,145.52,136.68,135.12,129.74,128.85,1 27.77,127.36,126.80,126.24,126.05,125.86,125.44,101.32,53.75,38.05,34.20,29.14,28.00,23.64.

[0206] Characterization data of compound 33: 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),10.29(s,1H),9.58(t,J=6.0Hz,1H),8.99(s,1H),8.38(s,1H),7.94–7.77(m,6H) ,7.60(m,2H),7.46(m,3H),7.33(t,J=7.5Hz,1H),3.58(mz,2H),3.40(m,4H),2.47(t,J=6.6Hz,2H),1.78–1.71(m,2H). 13 C NMR(101MHz,DMSO-d6)δ173.76,173.49,163.67,160.63,152.51,136.46,133.68,133.65,130.64,130.24,128.66,128 .29,128.13,127.97,127.87,127.27,126.97,125.52,121.46,117.59,103.13,51.84,39.26,35.47,34.17,28.65.HRMS m / z:calcd forC 28 H 28 ClN7O3S[M+H] + 578.1736found 578.1731.

[0207] Characterization data of compound 34: 1 H NMR(400MHz,DMSO-d6)δ10.20(s,1H),8.87(s,1H),8.82(t,J=5.7Hz,1H),8 .64(s,1H),8.02–7.95(m,3H),7.86(dd,J=5.9,3.5Hz,1H),7.62(s,1H),7. 55(m,4H),7.52–7.50(m,1H),7.41–7.34(m,2H),7.16(td,J=7.8,1.6Hz,1H ),3.53(s,2H),3.39(m,4H),2.44(t,J=6.8Hz,2H),1.77–1.71(m,2H).HRMS m / z:calcd forC 28 H 28 ClN7O3S[M+H] + 578.1736found 578.1754.

[0208] Characterization data of compound 35: 1H NMR (400MHz, DMSO-d6) δ9.66(s,1H),8.62(t,J=5.7Hz,1H),8.57(s,1H),8.36(s,1H),7.93(dd,J=8.2,1.6Hz,1H),7.61(s,1H),7. 50–7.28(m,12H),7.13(td,J=7.7,1.6Hz,1H),3.56(d,J=11.7Hz,2H),3.35(s,4H),2.45(t,J=6.7Hz,2H),1.73–1.66(m,2H).HRMS m / z:calcd for C 25 H 28 ClN7O3S[M+H] + 604.1892found604.1900.

[0209] Characterization data of compound 36: 1 H NMR(400MHz,DMSO-d6)δ9.91(s,1H),8.6–8.65(m,2H),8.62(s,1H),7.97(dd,J=8.2,1 .6Hz,1H),7.65(s,1H),7.50(dd,J=8.0,1.5Hz,1H),7.45(s,1H),7.36(dd,J=7.5,1.5 Hz,1H),7.15(td,J=7.7,1.6Hz,1H),6.98(d,J=2.3Hz,2H),6.25(t,J=2.3Hz,1H),3.7 3(s,6H),3.55(s,2H),3.39(q,J=6.4Hz,4H),2.47(t,J=6.7Hz,2H),1.78–1.72(m,2H). 13 C NMR(101MHz,DMSO-d6)δ173.86,166.07,161.71,160.79,160.69,157.77,141.13,136.60,129 .75,127.74,126.90,125.99,125.55,101.74,99.15,96.10,55.59,38.09,34.37,29.16.HRMS m / z:calcd for C 26 H 30 ClN7O5S[M+H] + 588.1790found 588.1794.

[0210] Characterization data of compound 37: 11H NMR (400 MHz, DMSO-d6) δ 12.25 (br s, 1H), 8.89 (t, J = 5.8 Hz, 1H), 8.63 (s, 1H), 8.59 (d, J = 7.7 Hz, 1H), 8.51 (s, 1H), 8.00 (dd, J = 8.2, 1.6 Hz, 1H), 7.58 (s, 1H), 7.48 (dd, J = 8.0, 1.4 Hz, 1H), 7.45–7.29 (m, 6H), 7.24–7.20 (m, 1H), 7.11 (dd, J = 8.0, 1.4 Hz, 1H), 5.15–5.08 (m, 1H), 3.55 (s, 2H), 3.37–3.29 (m, 4H), 2.46 (t, J = 6.7 Hz, 2H), 1.74–1.67 (m, 2H), 1.46 (d, J = 7.1 Hz, 3H) ppm. 13 13C NMR (101 MHz, DMSO-d6) δ 173.80, 166.33, 161.76, 160.50, 157.12, 145.47, 136.69, 129.71, 128.72, 127.72, 127.04, 126.45, 126.40, 125.48, 125.20, 101.26, 55.38, 48.50, 37.98, 34.25, 29.23, 28.93, 22.79 ppm. HRMS m / z: calcd for C 26 H 30 ClN7O3S [M+H] + 556.1892 found 556.1891.

[0211] Characterization data of Compound 38: 1 1H NMR (400 MHz, DMSO-d6) δ 8.89 (t, J = 5.7 Hz, 1H), 8.63 (s, 1H), 8.58 (d, J = 7.8 Hz, 1H), 8.47 (s, 1H), 8.02 (dd, J = 8.2, 1.6 Hz, 1H), 7.59 (t, J = 5.4 Hz, 1H), 7.47 (dd, J = 8.0, 1.5 Hz, 1H), 7.42–7.30 (m, 6H), 7.26–7.18 (m, 1H), 7.11 (td, J = 7.7, 1.6 Hz, 1H), 5.17–5.07 (m, 1H), 3.56 (s, 2H), 3.41–3.28 (m, 4H), 2.46 (t, J = 6.8 Hz, 2H), 1.75–1.68 (m, 2H), 1.46 (d, J = 7.1 Hz, 3H) ppm. 13C NMR(101MHz,DMSO-d6)δ173.85,166.35,161.79,160.50,157.11,145.45,136.70,129.69,128.71,127.73, 127.04,126.46,126.29,125.35,125.14,101.32,55.36,48.50,38.01,34.40,29.14,29.07,22.76ppm.HRMS m / z:calcd for C 26 H 30 ClN7O3S[M+H] + 556.1892found 556.1874.

[0212] Characterization data of compound 39: 1 H NMR (400MHz, DMSO-d6) δ8.87(t,J=5.8Hz,1H),8.64(s,1H),8.52(d,J=10.0Hz,2H),7.9 9(d,J=8.1Hz,1H),7.62(s,1H),7.48(d,J=8.0Hz,1H),7.42(t,J=6.0Hz,1H),7.38–7.3 0(m,5H),7.22(t,J=7.2Hz,1H),7.12(t,J=7.7Hz,1H),4.85(q,J=7.8Hz,1H),3.54(s,2 H),3.36–3.28(m,4H),2.45(t,J=6.8Hz,2H),1.91–1.70(m,4H),0.90(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO)δ166.65,161.75,160.52,157.04,144.53,136.69,129.71,128.68,127.73,1 27.12,127.00,126.44,125.52,125.23,101.32,54.94,37.97,34.46,29.45,29.11,11.87.HRMS m / z:calcd for C 27 H 32 ClN7O3S[M+H] + 570.2049found 570.2051.

[0213] Characterization data of compound 41: 11H NMR (400 MHz, DMSO-d6) δ 12.25 (s, 1H), 8.89 (t, J = 5.7 Hz, 1H), 8.65 (s, 1H), 8.55–8.45 (m, 2H), 8.01 (dd, J = 8.3, 1.5 Hz, 1H), 7.57 (s, 1H), 7.48 (dd, J = 8.0, 1.5 Hz, 1H), 7.34 (dt, J = 17.3, 7.5 Hz, 6H), 7.25–7.20 (m, 1H), 7.12 (td, J = 7.7, 1.6 Hz, 1H), 4.87–4.85 (m, 1H), 3.57 (s, 2H), 3.39–3.29 (m, 4H), 2.47 (t, J = 6.8 Hz, 2H), 1.89–1.69 (m, 4H), 0.90 (t, J = 7.3 Hz, 3H). 13 13C NMR (101 MHz, DMSO) δ 173.83, 166.65, 161.75, 160.50, 157.03, 144.53, 136.69, 129.70, 128.67, 127.72, 127.11, 127.00, 126.38, 126.34, 125.45, 125.19, 101.34, 54.95, 37.98, 34.2, 29.46, 29.11, 21.57, 11.87. HRMS m / z: calcd for C 27 H 32 ClN7O3S [M + H] + 570.2049 found 570.2052.

[0214] Characterization data of compound 42: 1 1H NMR (400 MHz, DMSO-d6) δ 8.96 (t, J = 5.7 Hz, 1H), 8.50 (s, 1H), 8.46 (s, 1H), 8.07–7.99 (m, 2H), 7.62 (s, 1H), 7.48 (dd, J = 8.0, 1.5 Hz, 1H), 7.43 (t, J = 5.5 Hz, 1H), 7.35 (td, J = 7.9, 1.5 Hz, 1H), 7.27 (t, J = 7.5 Hz, 2H), 7.22–7.08 (m, 4H), 3.98 (q, J = 6.9 Hz, 1H), 3.57 (s, 2H), 3.41–3.35 (m, 4H), 2.64–2.58 (m, 2H), 2.46 (t, J = 6.7 Hz, 2H), 1.89–1.81 (m, 1H), 1.76–1.72 (m, 2H), 1.16 (d, J = 6.5 Hz, 3H). 13C NMR (101MHz, DMSO) δ166.48,161.77,160.43,156.74,142.39,136.73,129.70,128.73,127.73, 126.24,126.12,125.31,125.10,101.68,44.64,38.18,38.01,34.42,32.58,29.17,21.24.HRMS m / z:calcdfor C 28 H 34 ClN7O3S[M+H] + 584.2205found 584.2204.

[0215] Characterization data of compound 43: 1 H NMR (400MHz, DMSO-d6) δ12.56(s,2H),8.75(s,1H),8.56(s,1H),8.47(d,J=15.4H z,2H),7.94(d,J=7.4Hz,1H),7.58(s,1H),7.48(d,J=7.6Hz,1H),7.41(s,1H),7.3 0(m,5H),7.19(s,1H),7.13(d,J=8.4Hz,1H),4.51(m,1H),3.55(s,2H),3.35(m,3 H),3.20–3.14(m,2H),3.01(m,1H),2.46(t,J=7.2Hz,2H),1.70–1.67(m,2H).HRMS m / z:calcd for C 27 H 30 ClN7O5S[M+H] + 600.1790found 600.1782.

[0216] Characterization data of compound 44: 1 H NMR(400MHz,DMSO-d6)δ12.28(s br,1H),8.82–8.76(m,2H),8.63(s,1H),8.59(s,1H),7.98(dd,J=8.1,1.6Hz,1H),7.59(s,1H),7.49–7.45(m,3H),7.43–7.33(m,5H),7.1 3(td,J=7.7,1.6Hz,1H),5.48(d,J=7.0Hz,1H),3.57(s,2H),3.37(q,J=6.8Hz,4H),2.47(t,J=6.8Hz,2H),1.77–1.70(m,2H),1.38(s,9H). 13C NMR(101MHz,DMSO-d6)δ173.87,170.15,167.17,161.70,160.63,157.94,137.01,136.64,129.73,128.95 ,128.56,128.52,127.72,126.61,125.67,125.35,100.68,81.60,57.57,38.02,34.23,29.15,28.05.HRMS m / z:calcd for C 30 H 36 ClN7O5S[M+H] + 642.2260found642.2260.

[0217] Characterization data of compound 45: 1 H NMR (400MHz, DMSO-d6) δ8.81(d,J=6.8Hz,2H),8.64(s,1H),8.58(s,1H),8.00(dd,J=8.2,1.6Hz,1H),7.64(s,1H),7.49–7.45(m,3H),7.44–7.30(m ,5H),7.12(td,J=7.7,1.6Hz,1H),5.49(d,J=7.0Hz,1H),3.58(s,2H),3.3 8(q,J=6.8Hz,4H),2.48(t,J=6.8Hz,1H),1.78–1.71(m,2H),1.38(s,9H). 13 C NMR (101MHz, DMSO) δ173.87,170.16,167.17,161.71,160.61,157.92,137.02,136.62,129.72,128.95,128.55, 128.51,127.72,127.41,126.51,125.57,125.30,100.71,81.61,57.58,49.09,38.04,34.35,29.17,28.04.HRMS m / z:calcd for C 30 H 36 ClN7O5S[M+H] + 642.2260found 642.2261.

[0218] Characterization data of compound 46: 11H NMR (400 MHz, DMSO-d6) δ 9.00 (s, 1H), 8.94 (t, J = 5.8 Hz, 1H), 8.60 (s, 1H), 8.52 (s, 1H), 7.99 (d, J = 8.1 Hz, 1H), 7.58 (s, 1H), 7.48 (d, J = 8.0 Hz, 1H), 7.37 (dt, J = 15.8, 6.8 Hz, 2H), 7.28 (t, J = 7.6 Hz, 2H), 7.21–7.10 (m, 4H), 3.57–3.53 (m, 2H), 3.36–3.32 (m, 4H), 2.50–2.42 (m, 2H), 1.75–1.68 (m, 2H), 1.25 (d, J = 4.3 Hz, 4H). 13 13C NMR (101 MHz, DMSO) δ 173.72, 167.76, 161.83, 160.55, 157.15, 144.10, 136.67, 129.73, 128.51, 127.76, 126.51, 125.98, 125.56, 125.29, 125.03, 101.17, 43.80, 37.99, 34.54, 34.18, 34.11, 29.07, 18.66. HRMS m / z: calcd for C 27 H 30 ClN7O3S [M+H] + 568.1892 found 568.1886.

[0219] Characterization data of compound 47: 1 1H NMR (400 MHz, DMSO-d6) δ 8.97 (t, J = 5.7 Hz, 1H), 8.51 (d, J = 4.4 Hz, 2H), 8.47 (s, 1H), 8.01 (d, J = 8.1 Hz, 1H), 7.60 (s, 1H), 7.48 (d, J = 8.0 Hz, 1H), 7.42 (d, J = 5.6 Hz, 1H), 7.35 (t, J = 7.8 Hz, 1H), 7.28 (t, J = 7.5 Hz, 2H), 7.19–7.10 (m, 4H), 3.56 (s, 2H), 3.40–3.34 (m, 4H), 2.98–2.93 (m, 1H), 2.46 (t, J = 6.8 Hz, 2H), 2.07–2.02 (m, 1H), 1.77–1.70 (m, 2H), 1.36–1.31 (m, 1H), 1.23–1.19 (m, 1H). 13C NMR(101MHz,DMSO-d6)δ173.81,168.27,162.79,161.65,160.49,157.05,141.97,136.66,129.72,128.66 ,127.74,126.35,126.05,125.44,125.21,101.11,37.99,34.24,33.31,29.13,24.57,15.71,14.56.HRMS m / z:calcd for C 27 H 30 ClN7O3S[M+H] + 568.1892found 568.1893.

[0220] Characterization data of compound 48: 1 H NMR(400MHz,DMSO-d6)δ12.30(s br,1H),8.94(t,J=5.8Hz,1H),8.54–8.47(m,2H),8.46(s,1H),8.00(dd,J=8.2,1.5Hz,1H),7.64(s,1 H),7.48(dd,J=8.0,1.4Hz,1H),7.42(t,J=5.7Hz,1H),7.37–7.29(m,2H),7.26–7.20(m,1H),7.12(td ,J=7.8,1.6Hz,1H),7.07–7.02(m,1H),3.55(s,2H),3.37(q,J=7.2,6.8Hz,4H),2.96–2.91(m,1H),2. 45(t,J=6.7Hz,2H),2.09–2.04(m,1H),1.77–1.70(m,2H),1.38–1.33(m,1H),1.25–1.23(m,1H).HRMS m / z:calcd for C 27 H 28 ClF2N7O4S[M+H] + 604.1704found604.1719.

[0221] Characterization data of compound 49: 11H NMR (400 MHz, DMSO-d6) δ 12.28 (s, 1H), 8.37 (s, 2H), 7.98 (d, J = 6.4 Hz, 1H), 7.75 (s, 1H), 7.58 (s, 1H), 7.46 (d, J = 7.6 Hz, 1H), 7.42 (s, 1H), 7.36–7.22 (m, 5H), 7.19 (m, 1H), 7.11 (d, J = 8.1 Hz, 1H), 5.10 (t, J = 7.7 Hz, 1H), 3.90 (s, 1H), 3.76 (m, 1H), 3.57 (m, 2H), 3.29 (s, 2H), 2.46 (t, J = 6.8 Hz, 2H), 2.43–2.30 (m, 1H), 1.95–1.80 (m, 2H), 1.77–1.62 (m, 3H), 0.84 (m, 2H). HRMS m / z: calcd for C 28 H 32 ClN7O3S [M+H] + 582.2049 found 582.2026.

[0222] Characterization data of compound 51: 1 1H NMR (400 MHz, DMSO-d6) δ 12.22 (s br, 1H), 8.33 (s, 1H), 8.16 (s, 1H), 8.05 (dd, J = 8.1, 1.5 Hz, 1H), 7.77 (t, J = 5.7 Hz, 1H), 7.61 (t, J = 5.4 Hz, 1H), 7.47 (dd, J = 8.0, 1.5 Hz, 1H), 7.43 (t, J = 5.6 Hz, 1H), 7.37–7.30 (m, 5H), 7.24 (td, J = 5.6, 3.0 Hz, 1H), 7.10 (td, J = 7.7, 1.6 Hz, 1H), 3.92 (dd, J = 10.7, 7.4 Hz, 1H), 3.71–3.64 (m, 2H), 3.57 (s, 3H), 3.39 (m, 5H), 2.48 (t, J = 6.6 Hz, 2H), 2.27 (m, 1H), 2.07–1.96 (m, 1H), 1.78–1.72 (m, 2H). 13C NMR(101MHz,DMSO-d6)δ173.88,166.74,160.82,159.80,156.40,141.52,136.79,129.66,128.95,127.73 ,127.62,127.12,125.99,125.17,124.92,104.71,55.37,41.73,40.37,38.17,34.35,29.19,29.17.HRMS m / z:calcd for C 28 H 32 ClN7O3S[M+H] + 582.2049found 582.2024.

[0223] Characterization data of compound 53: 1 H NMR(400MHz, DMSO-d6)δ8.36(s,1H),8.16(s,1H),8.07–8.01(m,1H),7.78(t,J=5.7Hz,1H), 7.63(s,1H),7.47(dd,J=8.0,1.5Hz,2H),7.35–7.32(m,5H),7.26–7.22(m,1H),7.10(td,J= 7.7,1.6Hz,1H),3.91(dd,J=10.7,7.4Hz,1H),3.70–3.64(m,2H),3.61–3.51(m,3H),3.44–3 .35(m,5H),2.48(t,J=6.4Hz,2H),2.29–2.24(m,1H),2.06–1.96(m,1H),1.78–1.71(m,2H). 13 C NMR(101MHz,DMSO-d6)δ173.92,166.72,160.79,159.80,156.41,136.77,129.67,128.95,127.73,127.6 2,127.13,126.06,125.25,124.95,104.65,55.38,48.56,42.04,40.52,38.14,34.35,29.16,23.65.HRMS m / z:calcdfor C 28 H 32 ClN7O3S[M+H] + 582.2049found 582.2052.

[0224] Characterization data of compound 54: 11H NMR (400 MHz, DMSO-d6) δ 12.21 (s, 1H), 8.28 (s, 1H), 8.04 (dd, J = 8.2, 1.5 Hz, 1H), 7.91 (s, 1H), 7.58 (s, 1H), 7.48–7.41 (m, 2H), 7.32 (t, J = 7.6 Hz, 3H), 7.27–7.20 (m, 3H), 7.14 (t, J = 5.9 Hz, 1H), 7.08 (td, J = 7.7, 1.5 Hz, 1H), 4.08–4.00 (m, 2H), 3.55 (s, 2H), 3.36–3.32 (m, 4H), 2.99 (t, J = 12.4 Hz, 2H), 2.81–2.74 (m, 1H), 2.46 (t, J = 6.7 Hz, 2H), 1.95–1.90 (m, 1H), 1.78–1.69 (m, 4H), 1.63–1.55 (m, 1H). 13 13C NMR (101 MHz, DMSO-d6) δ 173.80, 166.79, 160.10, 159.83, 155.56, 143.83, 136.83, 129.65, 129.01, 127.74, 127.43, 127.07, 125.78, 124.93, 124.77, 105.23, 60.24, 55.38, 42.72, 38.16, 34.21, 32.05, 29.18, 25.94.HRMS m / z: calcd for C 29 H 34 ClN7O3S [M + H] + 596.2205 found 596.2203.

[0225] Characterization data of compound 55: 1 1H NMR (400 MHz, DMSO-d6) δ 12.27 (s, 1H), 8.28 (s, 1H), 8.05 (dd, J = 8.2, 1.5 Hz, 1H), 7.93 (s, 1H), 7.58 (s, 1H), 7.46 (dd, J = 8.0, 1.4 Hz, 1H), 7.42 (t, J = 5.6 Hz, 1H), 7.35–7.27 (m, 5H), 7.21–7.13 (m, 2H), 7.09 (td, J = 7.7, 1.6 Hz, 1H), 4.17 (d, J = 13.0 Hz, 2H), 3.56 (s, 2H), 3.74–3.32 (m, 4H), 3.02 (t, J = 12.7 Hz, 2H), 2.83–2.75 (m, 1H), 2.46 (t, J = 6.8 Hz, 2H, 2H), 1.81–1.59 (m, 6H). 13C NMR(101MHz,DMSO-d6)δ173.91,166.74,160.07,159.83,155.63,146.10,136.87,129.66,128.86,127.74 ,127.28,126.66,125.73,124.92,124.74,105.42,45.73,45.60,42.39,38.13,34.34,33.47,29.23.HRMS m / z:calcdfor C 29 H 34 ClN7O3S[M+H] + 596.2205found 596.2234.

[0226] Characterization data of compound 56: 1 H NMR (400MHz, DMSO-d6) δ12.25(s,1H),10.81(s,1H),9.00(t,J=5.7Hz,1H),8.43(d,J=4.7Hz,3H),8.05–8.00(m,1 H),7.64(s,1H),7.57(d,J=7.8Hz,1H),7.48(dd,J=8.0,1.5Hz,1H),7.42(s,1H),7.35(dd,J=8.6,6.5Hz,2H),7.1 8(d,J=2.0Hz,1H),7.12(dd,J=7.7,1.6Hz,1H),7.07(dd,J=7.7,1.2Hz,1H),7.01(dd,J=7.7,1.6Hz,1H),3.56(s, 2H), 3.49 (q, J = 7.0Hz, 2H), 3.39 (q, J = 6.2Hz, 4H), 2.94 (t, J = 7.5Hz, 2H), 2.46 (t, J = 6.7Hz, 2H), 1.79–1.72 (m, 2H). 13 C NMR(101MHz,DMSO-d6)δ167.03,161.80,160.42,156.71,136.73,129.69,127.75,126.18,125.26,125.08,123.1 3,121.38,118.71,112.36,111.86,101.67,65.49,55.37,45.95,38.04,34.45,29.21,25.66,14.01,10.34.HRMS m / z:calcd for C 28 H 31 ClN8O3S[M+H] + 595.2001found 595.1979.

[0227] Characterization data of compound 57: 1 H NMR (400MHz, DMSO-d6) δ8.99(t,J=5.7Hz,1H),8.65(s,1H),8.46(s,1H),8.21(d,J=8.5Hz,1H),8.05(dd, J=8.2,1.6Hz,1H),7.67(t,J=5.3Hz,1H),7.56–7.43(m,2H),7.36(td,J=7.9,1.5Hz,1H),7.28–7.19(m,4 H),7.12(td,J=7.7,1.6Hz,1H),5.42(dd,J=8.5,5.1Hz,1H),4.51(td,J=5.1,1.9Hz,1H),3.58(s,2H),3. 42(q,J=6.1Hz,4H),3.10(dd,J=16.2,5.2Hz,1H),2.89(m,1H),2.48(t,J=6.8Hz,2H),1.80–1.73(m,2H). 13 C NMR (101MHz, DMSO) δ173.77,167.48,161.83,160.48,157.57,142.25,141.40,136.74,129.71,127.79,127.75,126 .75,126.22,125.29,125.24,125.10,124.94,101.57,72.64,57.50,49.08,45.94,38.11,34.33,29.25,9.78.HRMS m / z:calcdfor C 27 H 30 ClN7O4S[M+H] + 584.1841found 584.1859.

[0228] Characterization data of compound 58: 11H NMR (400 MHz, DMSO-d6) δ 10.84 (s, 1H), 8.83 (d, J = 6.0 Hz, 1H), 8.49 (s, 1H), 8.39 (s, 1H), 8.29 (d, J = 7.8 Hz, 1H), 7.97 (d, J = 7.9 Hz, 1H), 7.79 (d, J = 29.4 Hz, 1H), 7.58 (d, J = 8.0 Hz, 2H), 7.47 (dd, J = 8.1, 1.4 Hz, 1H), 7.36–7.30 (m, 2H), 7.19–7.16 (m, 1H), 7.11 (td, J = 7.7, 1.6 Hz, 1H), 7.04 (t, J = 7.5 Hz, 1H), 6.96 (t, J = 7.4 Hz, 1H), 4.49 (t, J = 10.2 Hz, 1H), 3.54 (s, 4H), 3.29–3.22 (m, 2H), 3.19–3.08 (m, 2H), 2.45 (t, J = 6.9 Hz, 2H), 1.73–1.66 (m, 2H). HRMS m / z: calcd for C 29 H 31 ClN8O5S [M+H] + 639.1899 found 639.1894.

[0229] Characterization data of compound 59: 1 1H NMR (400 MHz, DMSO-d6) δ 12.42 (s, 1H), 8.90 (t, J = 5.7 Hz, 1H), 8.65 (s, 1H), 8.57–8.46 (m, 2H), 8.02 (dd, J = 8.2, 1.5 Hz, 1H), 7.69 (t, J = 5.3 Hz, 1H), 7.47 (dd, J = 8.0, 1.5 Hz, 1H), 7.34 (dt, J = 15.0, 7.4 Hz, 5H), 7.26–7.14 (m, 2H), 7.11 (td, J = 7.7, 1.6 Hz, 1H), 4.89–4.83 (m, 1H), 3.70–3.59 (m, 2H), 3.43–3.29 (m, 4H), 1.89–1.68 (m, 4H), 1.10 (s, 6H), 0.90 (t, J = 7.3 Hz, 3H) ppm. 13C NMR(101MHz,DMSO-d6)δ176.38,170.80,166.65,161.76,160.49,157.02,144.52,136.69,129.69,128.66,127.71,127 .10,127.00,126.33,125.39,125.16,101.34,60.68,60.23,54.93,43.49,43.18,38.03,29.46,23.63,11.87ppm.HRMS m / z:calcd for C 29 H 36 ClN7O3S[M+H] + 598.2362found 598.2363.

[0230] Example 6. Synthesis of Compound 60

[0231] The synthesis method is as follows:

[0232]

[0233] Referring to the synthesis method of compound 21, intermediate 5c was used to replace 7c, and the raw material cyclopropylamine in the subsequent steps was replaced by (S)-α-methylbenzylamine to prepare the target compound 60. 1 H NMR (400MHz, DMSO-d6) δ8.89(t,J=5.7Hz,1H),8.62(s,1H),8.57(d,J=7.8Hz,1H),8.38(s,1H), 7.99(dd,J=8.2,1.6Hz,1H),7.64(dd,J=8.0,1.4Hz,1H),7.57(d,J=5.6Hz,1H),7.43–7.29(m,6H ),7.28–7.19(m,1H),7.05(td,J=7.7,1.6Hz,1H),5.12(p,J=7.1Hz,1H),3.63–3.50(m,2H),3.33 (dh,J=20.1,6.4Hz,4H),2.47(t,J=6.8Hz,2H),1.72(p,J=6.9Hz,2H),1.46(d,J=7.0Hz,3H)ppm. 13CNMR(101MHz,DMSO)δ166.35,161.79,160.52,157.13,145.45,137.97,132.88,128.71,128.36,127.0 4,126.46,125.69,125.65,117.43,101.31,55.37,48.50,41.79,38.01,34.28,29.12,22.76ppm.HRMS m / z:calcd for C 26 H 30 BrN7O3S[M+H] + 600.1387found600.1395and 602.1375.

[0234] Example 7. Synthesis of Compounds 61 and 62

[0235] The synthetic route is as follows: R is derived from different amine raw materials

[0236]

[0237] 1. Synthesis of Compound 61

[0238] Referring to the synthesis method of compound 23, the initial raw material 2,4-dichloro-5-pyrimidinecarboxylic acid ethyl ester was replaced with 2,4-dichloro-6-pyrimidinecarboxylic acid ethyl ester (61a) to prepare intermediate 61e, and then the raw material n-propylamine was replaced with benzylamine to continue to prepare the target compound 61 according to the synthesis method of compound 23. 1 H NMR (400MHz, DMSO-d6) δ8.83(t,J=6.3Hz,1H),8.28(d,J=8.2Hz,1H),7.96(s,1H),7.79(s,2H),7.52(s,1H),7.46(dd,J=8.0,1.5Hz,1H),7.40–7.20( m,7H),7.04(td,J=7.7,1.6Hz,1H),6.64(s,1H),4.47(d,J=6.3Hz,2H),3.5 5(d,J=6.5Hz,2H),3.38–3.30(m,4H),2.42(s,2H),1.80–1,73(m,2H).HRMS m / z:calcdfor C 25 H 28 ClN7O3S[M+H] + 542.1736found 542.1740.

[0239] 2. Synthesis of Compound 62

[0240] Referring to the synthesis method of compound 61, the target compound 62 can be obtained by replacing the raw material benzylamine with (S)-α-methylbenzylamine. 1 H NMR (400MHz, DMSO-d6) δ8.53(d,J=8.5Hz,1H),8.17(d,J=9.1Hz,2H),7.75(t,J=5. 3Hz,1H),7.67(t,J=5.4Hz,1H),7.52–7.45(m,2H),7.40–7.30(m,5H),7.28–7.24(m ,1H),7.07(td,J=7.7,1.6Hz,1H),6.60(s,1H),5.12–5.05(m,1H),3.56(s,2H),3.3 3(q,J=6.7Hz,2H),2.47(t,J=6.8Hz,2H),1.79–1.72(m,2H),1.48(d,J=6.9Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ173.77,164.40,163.07,159.43,155.25,144.32,137.13,129.67,128.91,127.77,1 27.43,126.49,124.74,124.14,123.86,96.71,60.23,48.62,38.49,34.32,29.16,22.65,21.23,14.56.HRMS m / z:calcd for C 26 H 30 ClN7O3S[M+H] + 556.1892found 556.1889.

[0241] Example 8. Synthesis of Compound 63

[0242] The synthesis method is as follows:

[0243]

[0244] Compound 53 (1 eq) was dissolved in acetonitrile, and triethylamine (1:5 volume ratio to acetonitrile) was added with stirring. Then, bromomethyl acetate (1.5 eq) was slowly added dropwise and reacted at 60°C for 30 h. The reaction solution was diluted with saturated sodium bicarbonate solution and extracted with ethyl acetate (20 ml, three times). The organic layers were combined, dried over anhydrous magnesium sulfate, concentrated, and purified by thin-layer chromatography (DCM:MeOH = 40:1) to obtain the target compound 63 as a white solid (76%). 1H NMR (400MHz, DMSO-d6) δ8.30(s,1H),8.16(s,1H),8.07(d,J=8.2Hz,1H),7.75(t,J=5.8Hz,1H),7. 47(d,J=8.0Hz,1H),7.41–7.18(m,7H),7.09(t,J=7.7Hz,1H),5.83(s,2H),4.22(t,J=7.3Hz,2H),3 .91(dd,J=10.7,7.5Hz,1H),3.75(t,J=6.7Hz,2H),3.66(d,J=7.0Hz,2H),3.54(s,1H),3.43–3.36( m,3H),2.79(t,J=6.7Hz,2H),2.30–2.23(m,1H),2.04(s,3H),2.01-2.96(m,1H),1.89–1.82m,2H). 13 C NMR (101MHz, DMSO) δ183.28,170.65,167.02,166.73,160.79,159.73,156.41,136.75,129.66,128.94,12 7.76,127.62,127.12,125.76,124.97,124.81,104.69,77.59,44.82,44.44,38.02,31.36,28.00,21.06.

[0245] The beneficial effects of the present invention are demonstrated below through specific test examples.

[0246] Test Example 1: Inhibitory activity of the compounds of the present invention on SIRT5 protein

[0247] (1) Experimental materials

[0248] Recombinant human SIRT5 protein (method see: Eur. J. Med. Chem. (2020) 112201); fluorescent peptide substrate P16 (Ac-Leu-Gly-Ser-Lys (Su) -AMC, customized by Dangang Biotechnology Co., Ltd.); Sigma's positive control product Suramin; all synthetic target compounds.

[0249] (2) Experimental methods

[0250] For specific operation methods, please refer to Eur.J.Med.Chem.(2020)112201; 60 μL of sample was added to each well of the test plate, including SIRT5 (0.2 μM), P16 (5 μM), NAD +(200 μM) and different concentrations of compounds and 150 mM NaCl, 25 mM Tris (pH 8.0) and 10% glycerol were added, and three parallel experiments were set up for all test wells. The dose-effect relationship obtained by the test was fitted with the corresponding half-maximal inhibitory effective concentration (IC 50 ).

[0251] (3) Experimental results

[0252] The inhibitory activity of the compounds of the present invention against SIRT5 was tested by the above experimental methods. The effective concentration (IC50) of the compounds against SIRT5 was 50 )See Table 1.

[0253] Table 1. Inhibitory activity of the compounds of the present invention on SIRT5

[0254]

[0255]

[0256] The above results show that the compounds of the present invention have good inhibitory activity against SIRT5, especially compounds 37, 41, 51 and 53. 50 The value was less than 1 μM, indicating excellent inhibitory activity.

[0257] The present invention provides a class of compounds with SIRT5 protein inhibitory activity, which can be used to prepare SIRT5 protein inhibitors and drugs for treating cancer. The present invention provides more options for the development and application of SIRT5 small molecule inhibitors and anticancer drugs.

Claims

1. A compound or a salt thereof, characterized in that: The structure of the compound is shown in Formula IVb: in, m is 1.

2. A compound or a salt thereof, characterized in that: The compound is one of the following compounds:

3. Use of the compound or salt thereof according to claim 1 or 2 in the preparation of a SIRT5 protein inhibitor.

4. A drug, characterized in that: The preparation is prepared by taking the compound or salt thereof as claimed in claim 1 or 2 as active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

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