HIV-1 (human immunodeficiency virus-1) non-nucleoside reverse transcriptase inhibitor as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an HIV-1 (human immunodeficiency virus-1) non-nucleoside reverse transcriptase inhibitor as well as a preparation method and application thereof. The HIV-1 non-nucleoside reverse transcriptase inhibitor is a piperidine-hydroxymethyl-diarylpyrimidine derivative, has two structural formulae, and also comprises a pharmaceutical salt, a hydrate, a solvate, a polycrystalline or a eutectic of the HIV-1 non-nucleoside reverse transcriptase inhibitor. The compound or the composition thereof can be used for preparing medicines for preventing or treating AIDS and the like. The result of an in-vitro cellular level anti-HIV-1 activity experiment shows that the micromolecule has relatively strong anti-HIV-1 biological activity, remarkably inhibits virus replication in MT-4 cells infected by an HIV-1 virus, and has relatively low cytotoxicity.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to an HIV-1 non-nucleoside reverse transcriptase inhibitor and a preparation method and use thereof. Background Art

[0002] AIDS is an infectious disease that poses a serious threat to global human health. It is known for its long incubation period, high mortality rate and inability to be completely cured, so it has always been the focus of research and attention. In 1981, the first case of AIDS was recorded in the United States, and two years later, in 1983, the HIV virus was identified as the pathogen of the disease. Since then, mankind's long struggle against AIDS has begun. According to statistics from the United Nations AIDS Program in 2023, the total number of people infected with HIV worldwide reached 39 million, of which about 1.3 million were newly infected in 2022, and the number of deaths caused by AIDS in the same year was 630,000.

[0003] After HIV invades the human body, it mainly attacks T lymphocytes, destroys the body's cellular and humoral immune mechanisms, and causes the loss of immune system function. The life cycle of HIV virus mainly includes the following steps: First, the virus binds and fuses with the host cell's T lymphocytes, releasing its own RNA genome into the host cell; then, the virus converts RNA into DNA through the reverse transcription process and integrates into the host cell's genome; then, it uses the host cell's enzymes and raw materials for transcription and translation to synthesize the genetic material and proteins required for viral replication; finally, the newly synthesized virus is released after assembly in the host cell, continues to infect other host cells, and further destroys the immune system. In the life cycle of HIV virus, reverse transcriptase (RT) plays a key role. It has three main functions: RNA-dependent DNA polymerase activity, DNA-dependent DNA polymerase activity, and RNA hydrolase activity. It is an important target for the development of HIV-1 inhibitors. Reverse transcriptase inhibitors are mainly divided into two categories: nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). NRTIs work by competing with the RT active site for substrates, but they have problems with poor selectivity and high toxicity. In contrast, NNRTIs bind to the allosteric binding pocket (NNIBP) about 10 angstroms near the RT active site in a non-competitive manner, and are characterized by high selectivity and high activity. Currently, the NNRTIs used in clinical practice are mainly second-generation drugs, such as the diaryl pyrimidine compounds rilpivirine (RPV) and etravirine (ETR). However, these drugs have problems such as poor water solubility (ETR has a solubility far below 1 μg / mL and RPV has a solubility of only 20 ng / mL), low patient response rates (ETR is 36.5% and RPV is 27.3%), and possible toxic side effects caused by long-term use, which limit their clinical application. In addition, the emergence of drug-resistant viral strains has also reduced the efficacy of these drugs. Therefore, the development of new and efficient NNRTIs with broad-spectrum resistance has become one of the hot areas of medicinal chemistry research.

[0004] The present invention aims to optimize the structure of RPV and ETR, enhance the interaction between the compound and the surrounding amino acids by introducing a hydroxymethyl group into the pyrimidine ring, improve the biological activity of the compound against drug-resistant virus strains, and improve its drugability. Summary of the invention

[0005] The purpose of the present invention is to provide a non-nucleoside reverse transcriptase inhibitor having strong anti-HIV-1 biological activity, which can significantly inhibit viral replication in MT-4 cells infected with HIV-1 virus and has low cytotoxicity, as well as a preparation method and use thereof.

[0006] The HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs) provided by the present invention contain piperidine-hydroxymethyl-diaryl pyrimidine derivatives, wherein the piperidine-hydroxymethyl-diaryl pyrimidine derivatives have the structural formula (I) or (II):

[0007]

[0008] Wherein, R is selected from but not limited to substituted or unsubstituted benzene ring;

[0009] As HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs), this class of compounds not only has strong biological activity, but also has low cytotoxicity and high selectivity.

[0010] The HIV-1 non-nucleoside reverse transcriptase inhibitors of the present invention also include pharmaceutically acceptable salts, stereochemical isomers, hydrates or solvates of the above derivatives.

[0011] In the present invention, the pharmaceutically acceptable salt is hydrochloride, hydrobromide, formate, methanesulfonate, trifluoromethanesulfonate, sulfate, phosphate, acetate, p-toluenesulfonate, tartrate, citrate, succinate, maleate, fumarate or malate.

[0012] The present invention also provides a method for preparing the above-mentioned HIV-1 non-nucleoside reverse transcriptase inhibitor containing piperidine-hydroxymethyl-diarylpyrimidine, and the specific steps are as follows:

[0013] Step 1: Use commercially available 2-thioether-4-chloro-5-ethylpyrimidine as a raw material and react with 4-bromo-2,6-dimethylphenol to obtain compound III, and the reaction formula is as follows:

[0014]

[0015] Specifically, a base is added to a solution containing 2-methylsulfide-4-chloro-5-ethylpyrimidine and 4,6-dimethylphenol. The mixture is stirred at a certain temperature. After the reaction is completed, the mixture is added dropwise to water. The filtered solid is dissolved in dichloromethane, washed with water and a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain a residue. The residue is then added dropwise to methanol to precipitate compound III as a yellow solid, which is dried in a vacuum oven and used in the next reaction.

[0016] Wherein: the solvent is selected from N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene, 1,2-dichloroethane;

[0017] The alkali is selected from sodium carbonate, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, potassium phosphate;

[0018] The molar ratio of the compound 2-thioether-4-chloro-5-ethylpyrimidine, 4-bromo-2,6-dimethylphenol, and base is 1:(1.0-3.0):(1-10), and the preferred ratio is 1:(1.0-1.2):(2-3);

[0019] The reaction temperature is 0 to 180°C, preferably 50 to 70°C;

[0020] The reaction time is 2 to 24 hours, preferably 6 to 10 hours.

[0021] Step 2: Use an oxidant to oxidize the methylthio group of compound III to sulfone to obtain compound IV, the reaction formula of which is as follows:

[0022]

[0023] Specifically, an oxidant is added to a solution of compound III and stirred at a certain temperature. After the reaction is completed, a reducing alkaline solution is added to the reaction solution. The organic phase is separated, washed with water and a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product IV as a light yellow solid, which is directly used in the next reaction without purification.

[0024] The solvent is selected from dichloromethane, ethyl acetate, methyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene;

[0025] The oxidant is selected from meta-chloroperbenzoic acid, hydrogen peroxide, peracetic acid, potassium monopersulfate (oxone), potassium persulfate;

[0026] The molar ratio of the compound III to the oxidant is 1:(2.0-20.0), and the preferred ratio is 1:(3.0-5.0);

[0027] The reducing alkaline solution is sodium thiosulfate, sodium sulfite, or sodium bisulfite;

[0028] The reaction temperature is -20 to 100°C, preferably 10 to 20°C;

[0029] The reaction time is 0.5 to 24 hours, preferably 1 to 2 hours;

[0030] Step 3: The methylthio group of compound IV undergoes a nucleophilic substitution reaction with 4-amino-1-Boc piperidine to obtain compound V. The reaction formula is as follows:

[0031]

[0032] Specifically, a base is added to a solution of compound IV and 4-amino-1-tert-butyloxycarbonyl-piperidine and stirred at a certain temperature. After the reaction is completed, the reaction mixture is filtered. The filtered off-white solid is placed in water and stirred. After filtering, the obtained solid is rinsed with water. The obtained solid is vacuum dried to obtain V as an off-white solid compound;

[0033] The solvent is selected from acetonitrile, dichloromethane, ethyl acetate, methyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene;

[0034] The alkali is selected from sodium carbonate, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, potassium phosphate;

[0035] The molar ratio of the compound IV and 4-amino-1-tert-butyloxycarbonyl-piperidine is 1:(1.0-10.0), preferably 1:(1.0-1.5);

[0036] The reaction temperature is 0 to 180°C, preferably 10 to 30°C;

[0037] The reaction time is 0.5 to 24 hours, preferably 8 to 10 hours.

[0038] Step 4: Reduce the ethyl ester group of compound V to a hydroxyl group to obtain compound VI. The reaction formula is as follows:

[0039]

[0040] Specifically, at a certain temperature, a reducing agent is added to a solution of compound V and stirred. After the reaction is completed, an alkaline solution is added to quench the reaction. Dichloromethane is then added for extraction. The organic phase is dried over anhydrous sodium sulfate and concentrated to obtain a crude product. Using ethyl acetate and petroleum ether as mobile phases, the compound VI is purified by silica gel column chromatography to obtain an off-white solid;

[0041] The solvent is selected from tetrahydrofuran, dichloromethane, dioxane, toluene, and 1,2-dichloroethane;

[0042] The reducing agent is selected from lithium aluminum tetrahydride, diisobutyl aluminum hydride, sodium borohydride, and tributyltin hydride;

[0043] The molar ratio of the compound V to the reducing agent is 1:(1.0-20.0), preferably 1:(2.0-5.0);

[0044] The reaction temperature is -78 to 100°C, preferably -10 to 10°C;

[0045] The reaction time is 0.5 to 16 hours, preferably 1 to 2 hours.

[0046] Step 5: The bromine of compound VI undergoes Suzuki coupling reaction with 4-cyanophenylboronic acid or pyridine-4-boronic acid to obtain compound VIIa or VIIb. The reaction formula is as follows:

[0047]

[0048] Specifically, compound VI, 4-cyanophenylboronic acid or pyridine-4-boronic acid and a base are added to a solvent. After adding a palladium catalyst, the mixture is degassed and replaced with nitrogen three times, and stirred at a certain temperature for reaction. After the reaction is completed, the reaction solution is concentrated. The obtained residue is purified by column chromatography and eluted with ethyl acetate and petroleum ether. The eluate is concentrated and vacuum dried to obtain compound VIIa or VIIb as an off-white solid;

[0049] The solvent is selected from acetonitrile, dichloromethane, ethyl acetate, methyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene, 1,2-dichloroethane;

[0050] The base is selected from sodium carbonate, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, potassium phosphate;

[0051] The molar ratio of the compound VI, 4-cyanophenylboronic acid or pyridine-4-boronic acid, palladium catalyst and base is 1:(1.0-10.0);(0.01%-1.0);(2.0-10.0), preferably 1:(1.0-1.5);(0.1%-0.2%);(2.0-3.0);

[0052] The reaction temperature is 50-180°C, preferably 100-120°C;

[0053] The reaction time is 2 to 48 hours, preferably 10 to 16 hours.

[0054] Step 6: Remove the Boc protecting group of compound VIIa or VIIb with acid to obtain compound VIIa or VIIb. The reaction formula is as follows:

[0055]

[0056] Specifically, a specific acid is added to a solution of compound VIIa or VIIb, and the reaction is stirred at a certain temperature. After the reaction is completed, the reaction solution is added dropwise to an alkaline solution to quench, and then extracted with dichloromethane. The organic phase is washed with water and a saturated sodium chloride solution, dried with anhydrous sodium sulfate and concentrated, and finally filtered to obtain an off-white solid compound VIIa or VIIb.

[0057] The solvent is selected from dichloromethane, dioxane, acetone, ethanol, methanol, isopropanol, tert-butanol, acetonitrile, tetrahydrofuran, toluene, and 1,2-dichloroethane;

[0058] The acid is selected from hydrochloric acid, hydrobromic acid, trifluoroacetic acid, sulfuric acid, and glacial acetic acid;

[0059] The reaction temperature is -50 to 100°C, preferably 0 to 30°C;

[0060] The reaction time is 0.5 to 48 hours, preferably 1 to 2 hours.

[0061] Step 7: The piperidine nitrogen atom of compound VIIIa or VIIIb reacts with a substituted benzyl halide to obtain compound I or II; the general reaction formula is as follows:

[0062]

[0063] Specifically, at a certain temperature, a base is added to a solvent containing compound VIIIa or VIIIb and a substituted benzyl halide, and the mixture is stirred at a certain temperature for several hours. After the reaction is completed, the reaction solution is poured into water and filtered to obtain a solid crude product; using dichloromethane and methanol as mobile phases, the final products I and II are purified by column chromatography, which are yellow or off-white solids (yield: 81-96%).

[0064] Wherein: the solvent used is one or more mixed solvents selected from N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, ethanol, isopropanol, n-butanol, and isobutanol;

[0065] The alkali used is one of the following: sodium carbonate, potassium carbonate, sodium hydroxide, potassium phosphate, one or more;

[0066] The substituted benzyl halide RCH2X is one of the following: chloro, bromo, iodo;

[0067] The molar ratio of the raw material compound VIIIa or compound VIIIb, substituted benzyl halide and base is 1:(1.0-2.0):(1-5). The optimal molar ratio is 1:1:2;

[0068] The reaction temperature is 0 to 180°C; preferably the reaction temperature is 20 to 50°C;

[0069] The reaction time is 2 to 24 hours, preferably 14 to 18 hours.

[0070] The present invention also provides a pharmaceutical composition, which contains an effective dose of the piperidine-hydroxymethyl-diaryl pyrimidine-containing non-nucleoside reverse transcriptase inhibitor, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0071] The present invention also provides the use of the piperidine-hydroxymethyl-diaryl pyrimidine-containing non-nucleoside reverse transcriptase inhibitor, or a pharmaceutically acceptable salt thereof, in the preparation of a drug for preventing and treating AIDS.

[0072] In the compounds of the present invention, a hydroxymethyl group is introduced on the 5-carbon of the pyrimidine nucleus, the purpose of which is to increase the antiviral activity of the target compound by increasing the hydrogen bond or polar interaction between the inhibitor and the surrounding amino acids (especially I180, E138K). At the same time, the biphenyl or benzene-pyridine structure on the left wing can penetrate into the binding pocket to enhance the binding force with the aromatic amino acids Phe227, Trp229, Tyr188 and Tyr181, further improving the biological activity of the target compound against drug-resistant virus strains. In addition, the more flexible 4-aminopiperidine fragment on the right wing increases the adaptability of the inhibitor in the non-nucleoside reverse transcriptase inhibitor binding pocket and the hydrogen bond interaction with the surrounding amino acids. The solvent / protein region of the benzyl targeting pocket of various substitutions is introduced on the piperidine nitrogen atom, further enhancing the interaction between the inhibitor and the reverse transcriptase. Therefore, a series of newly designed derivatives have a broad-spectrum inhibitory effect on HIV-1 mutants. Experimental results show that the compound has a more significant anti-HIV-1 activity, and has lower cytotoxicity and higher selectivity. DETAILED DESCRIPTION

[0073] The content of the present invention can be better understood through the following examples, but the content of the present invention cannot be limited thereto.

[0074] Example 1 to Example 26 are the preparations of compounds of formula (I). Example 27 to Example 37 are the preparations of compounds of formula (II).

[0075] Example 1: Synthesis of final product Ⅰa:

[0076]

[0077] Step 1: Use commercially available 2-thioether-4-chloro-5-ethylpyrimidine as a raw material and react with 4-bromo-2,6-dimethylphenol to obtain compound III, and the reaction formula is as follows:

[0078]

[0079] Potassium carbonate (50mmol, 2eq) was added to a DMF (100mL) solution containing 2-thioether-4-chloro-5-ethyl pyrimidine (50mmol, 1eq) and 4,6-dimethylphenol (10mmol, 1eq). The resulting mixture was stirred at 50°C for 16 hours. The reaction was detected by TLC until the raw material 2-thioether-4-chloro-5-ethyl pyrimidine disappeared. After the reaction was completed, the mixture was added dropwise to 1.5L of water and filtered. The obtained solid was dissolved in dichloromethane (1L); the organic phase was washed with water (500mL) and saturated sodium chloride (500mL), dried over anhydrous sodium sulfate, and concentrated to obtain about 100mL of residue; methanol was added dropwise to the residue to precipitate crystals to obtain compound III as a yellow solid, which was dried in a vacuum oven for the next reaction. Yield: 87%, mp: 116.2–118.9℃. 1 HNMR (400MHz, DMSO-d6): δ8.94(s,1H),7.40(s,2H),4.34(s,2H),2.26(s,3H),2.04(s,6H),1.32(s,3H).HRMS(ESI + ):m / z calcd.for C 16 H 18 BrN2O3S + [M+H] + :397.0216, found 397.0222.

[0080] Step 2: Use 85% by mass of m-chloroperbenzoic acid (m-CPBA) to oxidize the methylthio group of compound III into sulfone to obtain compound IV, the reaction formula of which is as follows:

[0081]

[0082] m-Chloroperbenzoic acid (85%, 150 mmol, 3 eq) was added in batches to a dichloromethane solution (1 L) of compound III (50 mmol, 1 eq), and the temperature of the reaction solution was controlled below 10°C during this process; the reaction solution was then stirred at 10°C for 2 hours; the reaction was detected by TLC until compound III disappeared; after the reaction was completed, saturated sodium carbonate solution (500 mL) and saturated sodium sulfite solution (500 mL) were added to the reaction solution; the organic phase was separated, washed with water (500 mL) and saturated sodium chloride solution (500 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product IV as a light yellow solid, which was directly used in the next step without purification.

[0083] Step 3: The methylthio group of compound IV undergoes a nucleophilic substitution reaction with 4-amino-1-Boc piperidine to obtain compound V, the reaction formula of which is as follows:

[0084]

[0085] Potassium carbonate (100 mmol, 2 eq) was added to a mixture containing compound IV (50 mmol, 1 eq), 4-amino-1-tert-butyloxycarbonyl-piperidine (50 mmol, 1 eq) and acetonitrile (50 mL); the reaction mixture was stirred at 25°C for 2 hours; the reaction was detected by TLC until compound IV disappeared; after the reaction was completed, the reaction mixture was filtered; the off-white solid obtained was placed in 500 mL of water and stirred for 30 min; filtered, and the filter cake was washed with water (100 mL); the obtained solid was vacuum dried to obtain compound V, which was an off-white solid compound. Yield: 82%, mp: 138.2–141.5°C. 1 H NMR (400MHz, DMSO-d6): δ8.73-8.70(m,1H),8.37-8.35(m,2H),4.29-4.22(m,2H),3.97-3.77(m,2H),3.06-2.71(m,2H),2. 39(s,1H),2.04-2.03(m,6H),1.73(d,J=16Hz,1H),1.50(d,J=16Hz,1H),1.37(s,9H),1.31-1.25(m,4H),1.15-1.05(m,1H). 13 C NMR (100MHz, DMSO-d6): δ167.1,163.7,162.6,154.2,149.6,133.4,131.0,1 17.8,100.5,99.5,79.2,60.4,49.7,48.5,42.5,31.0,28.5,16.2.HRMS(ESI + ):m / z calcd.for C 25 H 34 BrN4O5 + [M+H] + :549.1707,found 549.1714.

[0086] Step 4: The ethyl ester group of compound V is reduced to a hydroxyl group using lithium aluminum hydride to obtain compound VI; the reaction formula is as follows:

[0087]

[0088] Under nitrogen atmosphere at -10℃, lithium aluminum hydride (LiAlH4, 2.5mol / L tetrahydrofuran solution, 50mmol, 5eq) was gradually dripped into anhydrous tetrahydrofuran (50mL) solution of compound V (10mmol, 1eq); after reacting at -10℃ for 2 hours, the reaction was detected by TLC until compound V disappeared; at 0-5℃, sodium hydroxide aqueous solution (1.0mol / L) was added dropwise to quench the reaction; then dichloromethane (500mL) was added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain a crude product. Compound VI was purified by silica gel (200-300 mesh) column chromatography with ethyl acetate and petroleum ether as mobile phases to obtain compound VI as an off-white solid. Yield: 78%, mp: 141.2–143.8℃. 1 H NMR (400MHz, DMSO-d6): δ8.12(s,1H),7.33(s,2H),6.84(brs,1H),5.00(s,1H),4.46(s,2H) ,3.83(s,3H),3.02-2.74(m,2H),2.03(s,6H),1.71(s,2H),1.37(s,9H),1.29-1.14(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.3,161.4,158.9,154.3,149.5,133.7,131.2,117.7,109.7,79.0,56.3,48.2,43.1,31.5,28.5,16.3.HRMS (ESI + ):m / z calcd.for C 23 H 32 BrN4O4 + [M+H] + :507.1601, found 507.1600.

[0089] Step 5: The bromine of compound VI undergoes Suzuki coupling reaction with 4-cyanophenylboronic acid to obtain compound VIIa, and the reaction formula is as follows:

[0090]

[0091] Compound VI (10 mmol, 1 eq), 4-cyanophenylboronic acid (10 mmol, 1 eq) and cesium carbonate (20 mmol, 2 eq) were added to a mixed solvent of dioxane and water (150 mL and 50 mL, respectively); Pd(dppf)Cl2 (0.5 mmol, 0.05 eq) was added, the mixture was degassed and replaced with nitrogen three times; the reaction was stirred at 110°C for 16 hours; the reaction was monitored by TLC. The reaction solution was concentrated, and the residue was purified by silica gel (200-300 mesh) column chromatography, eluted with ethyl acetate and petroleum ether; the eluate was concentrated and vacuum dried to obtain compound VIIa as a colorless solid. Yield: 89%, mp: 194.9–197.4°C. 1 H NMR (400MHz, DMSO-d6): δ8.15(s,1H),7.92-7.87(m,4H),7.53(s,2H),6.86(brs,1H),5.02(t,J=4Hz,1H),4. 50(d,J=4Hz,2H),3.83(s,3H),3.01-2.73(m,2H),2.12(s,6H),1.70(s,2H),1.34(s,9H),1.25-1.17(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.4,158.8,154.3,150.9,144.7,135.5,133.3,131.8 ,127.8,127.6,119.4,110.2,109.7,79.0,56.4,48.5,43.0,31.5,28.5,16.8.HRMS(ESI + ):m / z calcd.for C 30 H 36 N5O4 + [M+H] + :530.2762, found 530.2745.

[0092] Step 6: Remove the Boc protecting group of compound VIIa with trifluoroacetic acid to obtain compound VIIa; the reaction formula is as follows:

[0093]

[0094] Trifluoroacetic acid (2.5 mL) was added dropwise to a solution of compound VIIa (2 mmol) in dichloromethane (5 mL) at 25°C; the reaction solution was stirred at 25°C for 1 hour; the reaction was monitored by TLC until compound VIIa disappeared; after the reaction was completed, the reaction solution was added dropwise to saturated sodium bicarbonate (50 mL) at 0°C to quench; it was extracted with dichloromethane (100 mL); the organic phase was dried over anhydrous sodium sulfate and concentrated to obtain compound VIIa as an off-white solid. Yield: 95%, mp: 245.1–247.5°C. 1 H NMR (400MHz, DMSO-d6): δ8.12(s,1H),7.92-7.86(m,4H),7.51(s,2H),6.82(brs,1H),5.01(s,1H),4.49(s, 2H),4.00-3.68(m,1H),2.79(s,2H),2.38(s,1H),2.12-2.04(m,7H),1.68-1.62(m,4H),1.23-1.16(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.1,160.2,157.3,149.7,144.5,135.5,131.8, 131.0,126.7,126.3,118.1,109.5,108.3,56.3,43.8,31.4,15.6.HRMS(ESI + ):m / z calcd.for C 25 H 28 N5O2 + [M+H] + :430.2238, found 430.2227.

[0095] Step 7: Compound VIIIa undergoes nucleophilic substitution reaction with 4-methylaminosulfonylbenzyl bromide. The reaction formula is as follows:

[0096]

[0097] At 25°C, potassium carbonate (1 mmol, 2 eq) was added to a DMF (1 mL) solution of compound VIIIa (0.5 mmol, 1 eq) and 4-methylaminosulfonylbenzyl bromide (0.5 mmol, 1 eq), and the mixture was stirred at 25°C for 16 hours. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was poured into water (50 mL). A solid crude product was obtained by filtration. The product Ia was purified by silica gel (200-300 mesh) column chromatography using dichloromethane and methanol as mobile phases to obtain an off-white solid.

[0098] Off-white solid,yield:85%,mp:210.1–211.9℃. 1 H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.92-7.86(m,4H),7.70(d,J=8Hz,2H),7.50(s,2H),7.41(d,J=8Hz,2H),7.23(q,J=4Hz,1H),6.70(d,J=4Hz,1 H),4.85(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.42(s,3H),2.67-2.65(m,2H ), 2.43 (d, J = 4Hz, 3H), 2.13 (s, 6H), 1.82-1.64 (m, 4H), 1.38-1.35 (m, 2H). 13 C NMR(100MHz,DMSO-d6):166.5,161.5,158.8,151.0,144.7,144.1,138.2,135.5,133.3,131.8, 129.6,127.8,127.5,127.1,119.4,110.2,109.7,62.0,56.4,52.8,48.5,31.6,29.1,16.7.HRMS calcd.for C 33 H 37 N6O4S + [M+H] + :613.2592, found613.2589.HPLC analysis:retention time=7.37min;peak area,98.89%(λ=254nm).IR:3470,3235,3140,2953,2224,1615cm -1 .

[0099] Example 2: Synthesis of Final Product Ib

[0100]

[0101] The operation is the same as in Example 1

[0102] Off-white solid,yield:83%,mp:192.1–194.1℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.91-7.85(m,4H),7.71(d,J=8Hz,2 H),7.48(s,2H),7.39(d,J=8Hz,2H),7.36-7.34(m,1H),6.68(d,J=4Hz,1H) ,4.86(t,J=4Hz,1H),4.54-4.51(m,3H),3.41-3.36(m,5H),2.84-2.80(m, 2H),2.67-2.64(m,2H),2.12(s,6H),1.81-1.63(m,4H),1.40-1.34(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.1,144.8,143.8,139.8,135.6,133.2,131.8,12 9.5,127.8,127.4,126.9,119.3,110.3,109.4,61.9,60.5,56.4,52.7,49.0,45.6,31.6,16.6.HRMS calcd.for C 34 H 39 N6O5S + [M+H] + :643.2697, found643.2677.HPLC analysis:retention time=7.31min;peak area,97.30%(λ=254nm).IR:3509,3268,3146,2944,2817,2229,1617,1405,1305cm -1 .

[0103] Example 3: Synthesis of Final Product Ic

[0104]

[0105] The operation is the same as in Example 1

[0106] Off-white solid,yield:87%,mp:120.5–122.9℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.91-7.85(m,4H),7.70(d,J=8Hz,2H),7 .48(s,2H),7.39(d,J=8Hz,2H),7.32(t,J=4Hz,1H),6.68(d,J=4Hz,1H),4.86(s ,1H),4.52(s,2H),4.27(s,1H),3.39-3.36(m,5H),2.84-2.79(m,2H),2.66-2.6 3(m,2H),2.12(s,6H),1.80-1.63(m,4H),1.57-1.50(m,2H),1.41-1.34(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.5,158.8,150.9,144.7,143.9,139.4,135.5,133.3,131.8,129.6 ,127.4,127.8,126.9,119.4,110.2,109.7,62.0,58.5,56.4,52.8,48.4,40.4,32.8,31.7,16.7.HRMS calcd.for C 35 H 41 N6O5S + [M+H] + :657.2854, found 657.2825.HPLC analysis:retention time=7.43min;peakarea,99.04%(λ=254nm).IR:3291,2922,2227,1614,1570,1154,1092cm -1 .

[0107] Example 4: Synthesis of Final Product Id

[0108] The operation is the same as in Example 1

[0109]

[0110] Off-white solid,yield:94%,mp:183.4–185.2℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.91-7.84(m,4H),7.72(d,J=8Hz,2H),7.5 1-7.50(m,1H),7.48(s,2H),7.39(d,J=8Hz,2H),6.69(d,J=4Hz,1H),4.86(t,J=4 Hz,1H),4.53(d,J=4Hz,2H),3.41(s,3H),3.31-3.28(m,2H),3.15(s,3H),2.95-2 .90(m,2H),2.67-2.64(m,2H),2.12(s,6H),1.82-1.63(m,4H),1.41-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.1,144.9,143.7,140.0,135.6,133.2,131.9,129.6 ,127.8,127.4,126.8,119.3,110.3,109.4,71.0,61.8,58.3,56.4,52.6,48.9,42.7,31.6,16.6.HRMS calcd.forC 35 H 41 N6O5S + [M+H] + :657.2854, found 657.2836.HPLC analysis:retention time=7.47min;peak area,97.62%(λ=254nm).IR:3512,3235,3139,2923,2226,1616,1524,1323,1155cm -1 .

[0111] Example 5: Synthesis of final product Ie

[0112]

[0113] The operation is the same as in Example 1.

[0114] Off-white solid,yield:85%,mp:215.9–217.6℃. 1H NMR (400MHz, DMSO-d6): 8.14 (s, 1H), 7.91-7.85 (m, 4H), 7.64 (d, J = 8Hz, 2H), 7 .48(s,2H),7.44(d,J=8Hz,2H),6.69(d,J=4Hz,1H),4.87(t,J=4Hz,1H),4.55( d,J=4Hz,1H),4.53(d,J=4Hz,2H),3.57-3.45(m,3H),3.18-3.15(m,3H),2.78 -2.72(m,2H),2.66(s,2H),2.13(s,6H),1.85-1.64(m,6H),1.48-1.36(m,4H). 13 CNMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.1,144.9,144.4,135.6,135.3,133.2,131.9,129. 7,127.8,127.7,127.4,119.3,110.3,109.4,64.4,61.8,56.4,52.7,48.9,43.6,33.5,31.6,16.6.HRMS calcd.for C 37 H 43 N6O5S + [M+H] + :683.3010, found 683.2984.HPLC analysis:retention time=7.45min;peak area,98.95%(λ=254nm).IR:3230,2944,2822,2772,2223,1613,1418cm -1 .

[0115] Example 6: Synthesis of Final Product If

[0116]

[0117] The operation is the same as in Example 1.

[0118] Off-white solid,yield:92%,mp:246.2–247.6℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.91-7.85(m,4H),7.76(d,J=8Hz,2H),7.49(s,2H),7.36(d,J=8Hz,2H),7.17(s,2H),6.68(d,J =4Hz,1H),4.84(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.38(s,3H),2.66-2.63(m,2H),2.13(s,6H),1.79-1.63(m,4H),1.40-1.34(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,144.9,143.4,143.3,135.6,133.2,131.9,12 9.3,127.8,127.4,126.1,119.3,110.3,109.4,62.0,56.5,52.7,49.0,31.7,16.7.HRMScalcd.for C 32 H 35 N6O4S + [M+H] + :599.2435, found 599.2428.HPLC analysis:retentiontime=7.26min;peak area,96.12%(λ=254nm).IR:3336,2930,2781,2225,1610,1418,1332cm -1 .

[0119] Example 7: Synthesis of final product IG

[0120]

[0121] The operation is the same as in Example 1.

[0122] Off-white solid,yield:93%,mp:210.6–212.4℃. 1 H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.91-7.83(m,6H),7.49-7.46(m,4H),6.70(d,J=4Hz,1H),4.86(t,J=4Hz,1 H),4.53(d,J=4Hz,2H),3.46(s,3H),3.16(s,3H),2.66(s,2H),2.13(s,6H),1.85-1.65(m,4H),1.42-1.36(m,2H).13 C NMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.1,145.3,144.8,140.1,135.6,133.2,131.9, 129.8,127.8,127.4,127.3,119.3,110.3,109.5,61.8,56.4,52.7,48.9,44.2,31.6,16.6.HRMS calcd.for C 33 H 36 N5O4S + [M+H] + :598.2483, found598.2466.HPLC analysis:retention time=7.32min;peak area,98.73%(λ=254nm).IR:3502,3247,2931,2231,1610cm -1 .

[0123] Example 8: Synthesis of Final Product Ih

[0124]

[0125] The operation is the same as in Example 1.

[0126] Off-white solid,yield:96%,mp:203.8–205.5℃. 1 H NMR (400MHz, DMSO-d6): δ8.19(q,J=4Hz,1H),8.13(s,1H),7.91-7.84(m,4H),7.73(d,J=8Hz,2H),7.48(s,2H),7.24(d,J=8Hz,2H),6.67(d,J=4Hz,1 H),4.83(t,J=4Hz,1H),4.52(d,J=4Hz,2H),3.35(s,3H),2.79(d,J=4Hz,3 H),2.65-2.62(m,2H),2.12(s,6H),1.77-1.61(m,4H),1.35-1.31(m,2H). 13C NMR (100MHz, DMSO-d6): δ167.1,166.5,161.7,159.2,151.1,144.9,142.3,135.6,133.9,133.2,131. 9,128.8,127.8,127.41,127.37,119.3,110.3,109.4,62.2,56.4,52.7,49.1,31.7,26.6,16.6.HRMS calcd.for C 34 H 37 N6O3 + [M+H] + :577.2922, found577.2910.HPLC analysis:retention time=7.14min;peak area,99.09%(λ=254nm).IR:3301,2925,2222,1634,1612,1568cm -1 .

[0127] Example 9: Synthesis of Final Product Ii

[0128]

[0129] The operation is the same as in Example 1.

[0130] Off-white solid,yield:85%,mp:204.2–205.9℃. 1 H NMR (400MHz, DMSO-d6): δ8.19 (t, J=

[0131] 4Hz,1H),8.14(s,1H),7.91-7.84(m,4H),7.76(d,J=8Hz,2H),7.48(s,2H),7.25(d,J=8Hz,2H),6.66(s,1H),4.86(s,1 H),4.58(s,1H),4.42(s,2H),3.54-3.35(m,7H),2.65-2.62(m,2H),2.12(s,6H),1.77-1.62(m,4H),1.39-1.30(m,2H). 13C NMR (100MHz, DMSO-d6): δ166.7,166.5,161.5,159.3,158.9,151.0,144.8,142.4,135.5,133.7,133.3 ,131.8,128.8,127.8,127.5,119.4,110.2,108.7,62.2,60.3,56.4,52.8,49.3,42.6,31.7,16.7.HRMS calcd.for C 35 H 39 N6O4 + [M+H] + :607.3027, found 607.3010.HPLC analysis:retention time=7.17min;peak area,96.86%(λ=254nm).IR:3561,3276,2920,2222,1611,1567cm -1 .

[0132] Example 10: Synthesis of Final Product Ij

[0133]

[0134] The operation is the same as in Example 1.

[0135] Off-white solid,yield:96%,mp:176.7–178.5℃. 1 H NMR (400MHz, DMSO-d6): δ8.25 (t, J=

[0136] 4Hz,1H),8.15(s,1H),7.92-7.85(m,4H),7.77(d,J=8Hz,2H),7.49(s,2H),7.27(d,J=8Hz,2H),6.70(d,J=4Hz,1H),4.8 7(s,1H),4.53(s,2H),4.36(s,1H),3.51-3.32(m,7H),2.68(s,2H),2.14(s,6H),1.84-1.65(m,6H),1.43-1.32(m,2H). 13C NMR (100MHz, DMSO-d6): δ166.7,166.5,161.6,159.2,151.1,144.9,141.9,135.6,134.1,133.2,131.9,12 9.0,127.9,127.5,127.4,119.3,110.3,109.4,62.0,59.3,56.4,52.6,48.9,37.1,32.9,31.5,16.7.HRMS calcd.for C 36 H 41 N6O4 + [M+H] + :621.3184, found 621.3177.HPLC analysis:retention time=7.16min;peak area,97.89%(λ=254nm).IR:3532,3250,2953,2229,1615cm -1 .

[0137] Example 11: Synthesis of Final Product Ik

[0138]

[0139] The operation is the same as in Example 1.

[0140] Off-white solid,yield:89%,mp:194.6–196.5℃. 1 H NMR (400MHz, DMSO-d6): δ8.27 (t, J=

[0141] 4Hz,1H),8.15(s,1H),7.90-7.84(m,4H),7.76(d,J=8Hz,2H),7.48(s,2H),7.25(d,J=8Hz,2H),6.67(d,J=4Hz,1H),4.87(t,J=4Hz,1H),4.53( d,J=4Hz,2H),3.47-3.43(m,4H),3.34(s,2H),3.27(s,3H),3.24(s,1H) ,2.65-2.62(m,2H),2.13(s,6H),1.77-1.62(m,4H),1.43-1.30(m,2H). 13C NMR (100MHz, DMSO-d6): δ166.7,166.5,161.5,158.8,150.9,144.8,142.4,135.5,133.5,133.3,131.8,12 8.9,127.8,127.5,127.3,119.4,110.2,109.6,71.0,62.2,58.4,56.4,52.8,48.6,39.3,31.6,16.7.HRMS calcd.for C 36 H 41 N6O4 + [M+H] + :621.3184, found 621.3169.HPLC analysis:retention time=7.33min;peak area,95.09%(λ=254nm).IR:3558,3277,2921,2222,1612,1568cm -1 .

[0144] Example 12: Synthesis of Final Product Il

[0145]

[0146] The operation is the same as in Example 1.

[0147] Off-white solid,yield:90%,mp:207.9–209.7℃. 1 H NMR (400MHz, DMSO-d6): δ8.15(s,1H),7.90-7.84(m,4H),7.80(d,J=8Hz,2H),7.67(s,1H),7.48(s,2H),7.26(d,J=8Hz,2H),7. 14(s,1H),6.68(s,1H),4.88(s,1H),4.53(s,2H),3.40(s,3H),2.67(s,2H),2.12(s,6H),1.84-1.65(m,4H),1.42-1.33(m,2H). 13 C NMR (100MHz, DMSO-d6): δ168.4,168.3,166.5,161.6,159.2,151.1,144.9,142.0,135.6,133.7, 133.2,131.9,129.0,127.9,127.4,119.3,110.3,109.4,62.0,56.4,52.5,48.9,31.5,16.6.HRMS calcd.forC 33H 35 N6O3 + [M+H] + :563.2765, found 563.2760.HPLC analysis:retention time=7.09min;peak area,99.17%(λ=254nm).IR:3355,3248,2946,2223,1667,1611,1586cm -1 .

[0148] Example 13: Synthesis of Final Product Im

[0149] The operation is the same as in Example 1.

[0150]

[0151] The operation is the same as in Example 1.

[0152] Off-white solid,yield:91%,mp:165.5–167.6℃. 1 H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.83(m,4H),7.74-7.72(m,3H),7.47(s,2H),7.35-7.14(m,3H),6.6 7(d,J=4Hz,1H),4.52(s,2H),3.37(s,4H),2.68-2.65(m,2H),2.12(s,6H),1.91-1.63(m,4H),1.42-1.32(m,2H). 13 CNMR (100MHz, DMSO-d6): δ168.4,166.5,161.5,158.8,150.9,144.8,139.2,135.5,134.7,133. 3,132.0,131.8,128.4,127.8,127.6,126.4,119.4,110.2,109.5,62.5,56.4,52.8,31.6,29.5 16.7.HRMS calcd.for C 33 H 35 N6O3 + [M+H] + :563.2765, found 563.2762. HPLC analysis: retention time=7.13min; peak area, 98.00% (λ=254nm).

[0153] Example 14: Synthesis of Final Product In

[0154]

[0155] The operation is the same as in Example 1.

[0156] Off-white solid,yield:95%,mp:253.0–254.9℃. 1 H NMR (400MHz, DMSO-d6): δ8.24(q,J=4Hz,1H),8.14(s,1H),7.90-7.83(m,4 H),7.69-7.67(m,2H),7.47(s,2H),7.35-7.28(m,2H),6.67(d,J=4Hz,1H), 4.86(t,J=4Hz,1H),4.53-4.52(d,J=4Hz,2H),3.35(s,3H),2.80(d,J=4Hz, 3H),2.66-2.64(m,2H),2.12(s,6H),1.78-1.62(m,4H),1.39-1.33(m,2H). 13 C NMR (100MHz, DMSO-d6): δ167.3,166.5,161.7,159.2,151.1,144.9,139.3,135.6,135.1,133.2,131.9,131 .7,128.4,127.9,127.8,127.4,126.0,119.3,110.3,109.4,62.5,56.4,52.7,49.1,31.7,26.6,16.6.HRMS calcd.for C 34 H 37 N6O3 + [M+H] + :577.2922, found577.2923.HPLC analysis:retention time=7.49min;peak area,99.66%(λ=254nm).IR:3384,3276,2939,2804,2225,1611,1588cm -1 .

[0157] Example 15: Synthesis of Final Product Io

[0158]

[0159] The operation is the same as in Example 1.

[0160] Off-white solid,yield:79%,mp:191.2–193.4℃.1 H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.86(m,4H),7.85(d,J=8Hz,2H),7.48(s,2H),7.34(d,J=8Hz,2H),6.70(t,J=4Hz,1H), 4.85(t,J=4Hz,1H),4.52(d,J=4Hz,2H),3.85(s,3H),3.43(s,3H),2.67(s,2H),2.12(s,6H),1.83-1.63(m,4H),1.38-1.35(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.6,166.5,161.7,159.2,151.1,144.8,135.6,133.2,131.9,129.5,129. 3,129.0,127.8,127.4,119.3,110.3,109.4,62.0,56.4,52.6,52.4,48.9,31.5,16.6.HRMScalcd.for C 34 H 36 N5O4 + [M+H] + :578.2762, found 578.2764.HPLC analysis:retention time=7.86min;peak area,99.22%(λ=254nm).IR:3507,3384,2948,2798,2225,1720,1614,1529,1290cm -1 .

[0161] Example 16: Synthesis of Final Product Ip

[0162]

[0163] The operation is the same as in Example 1.

[0164] Off-white solid,yield:85%,mp:168.3–170.4℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.84(m,4H),7.62(d,J=16Hz,1H) ,7.58(d,J=8Hz,2H),7.48(s,2H),7.22(d,J=8Hz,2H),6.68(d,J=4Hz,1H),6. 54(d,J=16Hz,1H),4.84(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.73(s,3H),3.33 (s,3H),2.65-2.63(m,2H),2.13(s,6H),1.77-1.62(m,4H),1.39-1.33(m,2H). 13 C NMR (100MHz, DMSO-d6): δ167.1,166.5,161.7,159.2,151.1,144.8,144.7,141.9,135.6,133.2,131.9, 129.6,128.6,127.8,127.4,119.3,117.9,110.3,109.4,62.2,56.4,52.7,51.8,49.1,31.7,16.6.HRMS calcd.for C 36 H 38 N5O4 + [M+H] + :604.2918, found 604.2909.HPLC analysis:retention time=8.03min;peak area,98.70%(λ=254nm).IR:3565,3355,3226,2945,2822,2228,1708,1614cm -1 .

[0165] Example 17: Synthesis of Final Product Iq

[0166]

[0167] The operation is the same as in Example 1.

[0168] Off-white solid,yield:89%,mp:238.7–240.9℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.84(m,4H),7.70(d,J=8Hz,2H),7.48(s,2H),7.38(d,J=8Hz,2H),6.68(d,J=4Hz,1 H),4.85(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.41(s,3H),2.64-2.61(m,2H),2.12(s,6H),1.81-1.62(m,4H),1.40-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,145.2,144.8,135.6,133.2,132.5,131.9,1 29.9,127.8,127.4,119.27,119.25,110.3,110.2,109.4,61.9,56.4,52.7,48.9,31.6,16.6.HRMS calcd.forC 33 H 33 N6O2 + [M+H] + :545.2660, found 545.2657.HPLC analysis:retention time=7.77min;peak area,96.88%(λ=254nm).IR:3544,3241,3132,2920,2790,2229,1615,1524cm -1 .

[0169] Example 18: Synthesis of Final Product Ir

[0170]

[0171] The operation is the same as in Example 1.

[0172] Off-white solid,yield:85%,mp:126.0–127.8℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.84(m,4H),7.66(d,J=8Hz,1H),7.60(s,1H),7.54-7.46(m,4H),6.69(d,J=4Hz,1 H), 4.86 (t, J = 4Hz, 1H), 4.53 (d, J = 4Hz, 2H), 3.40 (s, 3H), 2.66-2.64 (m, 2H), 2.13 (s, 6H), 1.96-1.64 (m, 4H), 1.41-1.32 (m, 2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.1,144.9,140.7,135.6,134.0,133.2,132.4,131.9,131.2 ,129.8,127.8,127.4,119.3,119.2,111.8,110.3,109.4,61.4,56.4,52.5,48.9,31.6,16.6.HRMScalcd.for C 33 H 33 N6O2 + [M+H] + :545.2660, found 545.2652.HPLC analysis:retention time=7.63min;peak area,99.58%(λ=254nm).IR:3243,2924,2226,1613,1416cm -1 .

[0173] Example 19: Synthesis of Final Product ls

[0174]

[0175] The operation is the same as in Example 1.

[0176] Off-white solid,yield:82%,mp:188.7–190.9℃. 1H NMR (400MHz, DMSO-d6): 8.15 (s, 1H), 7.88-7.83 (m, 4H), 7.72 (d, J = 8Hz, 1H), 7.60 (t, J = 8Hz, 1H), 7.48 (s, 2H), 7.43-7.40 (m, 2H), 6.70 (d ,J=4Hz,1H),4.86(t,J=4Hz,1H),4.54(d,J=4Hz,2H),3.50(s,3H),2.67-2.65(m,2H),2.13(s,6H),1.89-1.63(m,4H),1.42-1.30(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,144.9,142.8,135.6,133.32,133.25,133.2,131.9 ,130.3,128.3,127.8,127.4,119.3,118.0,112.6,110.3,109.4,60.3,56.4,52.6,48.9,31.6,16.6.HRMS calcd.for C 33 H 33 N6O2 + [M+H] + :545.2660, found 545.2657.HPLCanalysis:retention time=8.47min;peak area,98.33%(λ=254nm).IR:3490,3363,3243,2961,2825,2226,1614,1542cm -1 .

[0177] Example 20: Synthesis of Final Product It

[0178]

[0179] The operation is the same as in Example 1.

[0180] Yellow solid,yield:83%,mp:227.6–229.4℃. 1H NMR (400MHz, DMSO-d6): δ8.15(s,1H),8.12(d,J=8Hz,2H),7.90-7.84(m,4H),7.48-7.46(m,4H),6.70(d,J=4Hz,1H), 4.86(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.48(s,3H),2.67(s,2H),2.13(s,6H),1.86-1.64(m,4H),1.43-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,147.4,147.2,144.8,135.6,133.2,131 .9,130.0,127.8,127.4,123.7,119.3,110.3,109.4,61.5,56.4,52.7,48.9,31.6,16.6.HRMS calcd.for C 32 H 33 N6O4 + [M+H] + :565.2558, found 565.2553.HPLC analysis:retention time=8.02min;peak area,97.78%(λ=254nm).IR:3586,3249,2952,2750,2226,1607,1520cm -1 .

[0181] Example 21: Synthesis of Final Product Iu

[0182]

[0183] The operation is the same as in Example 1.

[0184] Yellow solid,yield:85%,mp:202.9–204.7℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),8.07-8.05(m,2H),7.89-7.83(m,4H),7.65-7.63(m,1H),7.58-7.54(m,1H),7.47(s,2H),6.69(d, J=4Hz,1H),4.86(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.48(s,3H),2.69-2.67(m,2H),2.12(s,6H),1.88-1.65(m,4H),1.42-1.37(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,148.5,144.9,141.5,135.6,133.2,131.9,130.0 ,127.8,127.4,123.4,122.3,119.3,110.3,109.4,61.3,56.4,52.5,48.9,31.6,16.6.HRMScalcd.for C 32 H 33 N6O4 + [M+H] + :565.2558, found565.2556.HPLC analysis:retention time=7.96min;peak area,99.02%(λ=254nm).IR:3501,3383,2918,2802,2757,2223,1611cm -1 .

[0185] Example 22: Synthesis of Final Product IV

[0186]

[0187] The operation is the same as in Example 1.

[0188] Off-white solid,yield:82%,mp:198.9–201.2℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.91-7.85(m,4H),7.48(s,2H),7.22-7.19(m,2H),7.08-7.03(m,2H),6.68(d,J=4Hz ,1H),4.85(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.32(s,3H),2.65(s,2H),2.12(s,6H),1.78-1.63(m,4H),1.36-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.8(d,J=241Hz),161.7,159.2,151.1,144.8,135.6,135.0,133.2,131.9 (d,J=9Hz),131.0,127.8,127.4,119.3,115.2(d,J=12Hz),110.3,109.4,61.6,56.4,52.5,49.0,31.6,16.6. 19 F NMR(376MHz,DMSO-d6):δ-116.1.HRMS calcd.for C 32 H 33 FN5O2 + [M+H] + :538.2613, found538.2613.HPLC analysis:retention time=7.69min;peak area,98.71%(λ=254nm).IR:3517,3386,2928,2769,2225,1612,1538cm -1 .

[0189] Example 23: Synthesis of Final Product Iw

[0190]

[0191] The operation is the same as in Example 1.

[0192] Off-white solid,yield:83%,mp:165.9–168.0℃. 1H NMR (400MHz, DMSO-d6): δ8.15(s,1H),7.90-7.84(m,4H),7.61(d,J=8Hz,2H),7.48(s,2H),7.42(d,J=8Hz,2H),6.70(d,J=4H z,1H),4.86(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.44(s,3H),2.67(s,2H),2.13(s,6H),1.84-1.64(m,4H),1.42-1.33(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,144.8,143.7,135.6,133.2,131.9,129.8,128.2(d,J=32H z),127.8,127.4,125.4(q,J=4Hz),124.8(q,J=270Hz),119.3,110.3,109.4,61.8,56.4,52.6,48.9,31.5,16.6. 19 F NMR(376MHz,DMSO-d6):δ-60.87.HRMS calcd.for C 33 H 33 F3N5O2 + [M+H] + :588.2581, found 588.2572.HPLC analysis:retention time=8.45min;peak area,98.84%(λ=254nm).IR:3502,3359,2946,2821,2225,1611,1541cm -1 .

[0193] Example 24: Synthesis of Final Product Ix

[0194]

[0195] The operation is the same as in Example 1.

[0196] Off-white solid,yield:84%,mp:197.3–199.6℃. 1H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.84(m,4H),7.48(s,2H),7.30(d,J=8Hz,2H),7.22(d,J=8Hz,2H),6.69(d,J=4H z,1H),4.85(t,J=4Hz,1H),4.53(d,J=4Hz,2H),3.36(s,3H),2.64(s,2H),2.13(s,6H),1.80-1.63(m,4H),1.40-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.1,147.8,144.8,138.3,135.6,133.2,131.9,130. 8,127.81,127.4,120.6(d,J=255Hz),120.9,119.3,110.3,109.4,61.5,56.4,52.6,49.0,31.6,16.6. 19 FNMR(376MHz,DMSO-d6):δ-56.8.HRMS calcd.for C 33 H 33 F3N5O3 + [M+H] + :604.2530, found604.2526.HPLC analysis:retention time=8.46min;peak area,99.21%(λ=254nm).IR:3366,2930,2761,2226,1615,1538cm -1 .

[0197] Example 25: Synthesis of Final Product Iy

[0198]

[0199] The operation is the same as in Example 1.

[0200] Off-white solid,yield:81%,mp:203.0–204.9℃. 1H NMR (400MHz, DMSO-d6): δ8.16 (s, 1H), 7.90-7.85 (m, 4H), 7.48 (s, 2H), 7.18 (d, J = 8Hz, 2H), 7.10 (d, J = 8Hz, 2H), 6.78 (d, J = 4Hz, 1H),4.92(s,1H),4.53(s,2H),3.61(s,3H),2.84(s,2H),2.28(s,3H),2.14-2.12(m,8H),1.76-1.73(m,2H),1.56-1.49(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.6,159.1,151.1,144.8,136.5,135.6,134.9,133.1,131.9,129. 3,129.1,127.8,127.3,119.3,110.3,109.4,62.0,56.4,52.3,48.9,31.3,21.1,16.6.HRMScalcd.for C 33 H 36 N5O2 + [M+H] + :534.2864, found 534.2856.HPLC analysis:retention time=7.98min;peak area,96.07%(λ=254nm).IR:3243,2924,2226,1613,1416cm -1 .

[0201] Example 26: Synthesis of Final Product Iz

[0202]

[0203] The operation is the same as in Example 1.

[0204] Off-white solid,yield:86%,mp:206.1–208.4℃. 1 H NMR (400MHz, DMSO-d6): δ8.14(s,1H),7.90-7.84(m,4H),7.48(s,2H),7.22(d,J=8Hz,2H),7.13(d,J=8Hz,2H),6.67(d,J=4 Hz,1H),4.88(s,2H),4.52-4.47(m,4H),3.61(s,3H),2.68-2.66(m,2H),2.12(s,6H),1.81-1.63(m,4H),1.40-1.34(m,2H).13 C NMR (100MHz, DMSO-d6): δ166.5,161.5,158.8,150.9,144.7,141.6,137.1,135.5,133.3,131.8, 129.0,127.8,127.5,126.8,119.4,110.2,108.7,63.2,62.4,56.4,52.6,48.5,31.6,16.7.HRMS calcd.for C 33 H 36 N5O3 + [M+H] + :550.2813, found 550.2818.HPLC analysis:retention time=7.22min;peakarea,97.42%(λ=254nm).IR:3534,3236,3138,2922,2816,2225,1617,1528,1417,1147cm -1 .

[0205] Example 27: Synthesis of Final Product IIa

[0206]

[0207] Step 1: The bromine of compound VI (in Example 1) undergoes Suzuki coupling reaction with pyridine-4-phenylboronic acid to obtain compound VIIb, the reaction formula of which is as follows:

[0208]

[0209] Compound VI (10 mmol, 1 eq), pyridine-4-boronic acid (10 mmol, 1 eq) and cesium carbonate (20 mmol, 2 eq) were added to a mixed solvent of dioxane and water (150 mL and 50 mL, respectively); Pd(dppf)Cl2 (0.5 mmol, 0.05 eq) was added, and the mixture was degassed and replaced with nitrogen three times. The reaction was stirred at 110°C for 16 hours; the reaction was monitored by TLC to be complete; the reaction solution was concentrated, and the resulting residue was purified by column chromatography, eluted with ethyl acetate and petroleum ether; the eluate was concentrated and vacuum dried to obtain compound VIIb as a colorless solid. Yield: 83%, mp: 190.2–192.8°C. 1H NMR (400MHz, DMSO-d6): δ8.62(d,J=4Hz,1H),8.14(s,1H),7.71(d,J=4Hz,2H),7.58(s,2H),6.87(brs,1H),5.02(t,J=4 Hz,1H),4.50(d,J=4Hz,2H),3.83(s,3H),3.00-2.74(m,2H),2.13(s,6H),1.70(s,2H),1.34(s,9H),1.25-1.17(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.4,158.8,154.3,151.2,151.1,150.7,147.6 ,134.4,131.9,127.3,121.5,79.0,56.4,48.5,42.6,31.5,28.5,16.7.HRMS(ESI + ):m / z calcd.for C 28 H 36 N5O4 + [M+H] + :506.2762, found 506.2755.

[0210] Step 2: The Boc protecting group of compound VIIb is removed with trifluoroacetic acid to obtain compound VIIIb, and the reaction formula is as follows:

[0211]

[0212] Trifluoroacetic acid (2.5 mL) was added dropwise to a solution of compound VIIb (2 mmol) in dichloromethane (5 mL) at 25°C; the reaction solution was stirred at 25°C for 1 hour; the reaction was monitored by TLC until compound VIIb disappeared. After the reaction was completed, the reaction solution was added dropwise to saturated sodium bicarbonate (50 mL) at 0°C to quench; and extracted with dichloromethane (100 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain an off-white solid compound VIIIb, which was directly used in the next step.

[0213] Step 3: Compound VIIIb undergoes nucleophilic substitution reaction with 4-methylaminosulfonylbenzyl bromide to obtain IIa. The reaction formula is as follows:

[0214]

[0215] At 25°C, potassium carbonate (1 mmol, 2 eq) was added to a DMF (1 mL) solution of compound VIIIb (0.5 mmol, 1 eq) and 4-methylaminosulfonylbenzyl bromide (0.5 mmol, 1 eq), and the mixture was stirred at 25°C for 16 hours. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was poured into water (50 mL). A solid crude product was obtained by filtration. The product IIa was purified by column chromatography using dichloromethane and methanol as mobile phases to obtain an off-white solid.

[0216] Off-white solid,yield:90%,mp:172.2–174.4℃. 1 H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.15(s,1H),7.70(d,J=8Hz,2H),7.67(d,J=4Hz,2H),7.53(s,2H),7.39(d,J=8Hz,2H),7.25(s, 1H),6.70(s,1H),4.86(s,1H),4.53(s,2H),3.39(s,3H),2.65(s,2H), 2.43-2.42(m,3H),2.13(s,6H),1.81-1.65(m,4H),1.38-1.32(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.4,150.6,147.2,144.0,138.6,134.5,131. 9,129.5,127.2,127.0,121.5,109.4,61.9,56.4,52.7,49.0,31.7,29.1,16.6.HRMScalcd.for C 31 H 37 N6O4S + [M+H] + :589.2592, found589.2588.HPLC analysis:retention time=6.27min;peak area,99.64%(λ=254nm).IR:3253,3139,2954,2776,1616,1149cm -1 .

[0217] Example 28: Synthesis of Final Product IIb

[0218]

[0219] The operation is the same as Example 27.

[0220] Off-white solid,yield:82%,mp:214.1–216.5℃. 1 H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.15(s,1H),7.71(d,J=8Hz,2H) ,7.68(d,J=4Hz,2H),7.54(s,2H),7.39(s,1H),7.37(d,J=8Hz,2H),6.71(d,J= 4Hz,1H),4.86(t,J=4Hz,1H),4.53-4.52(m,3H),3.41-3.37(m,5H),2.85-2.80 (m,2H),2.65-2.63(m,2H),2.14(s,6H),1.80-1.63(m,4H),1.37-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.5,159.0,151.2,150.6,147.1,143.9,139.5,134.4,13 1.9,129.6,127.4,126.9,121.5,109.7,62.0,60.4,56.4,52.8,48.6,45.5,31.6,16.7.HRMS calcd.for C 32 H 39 N6O5S + [M+H] + :619.2697, found619.2686.HPLC analysis:retention time=5.90min;peak area,98.73%(λ=254nm).IR:3386,3243,2930,2827,2777,1608,1520,1412,1159cm -1 .

[0221] Example 29: Synthesis of Final Product IIc

[0222]

[0223] The operation is the same as Example 27.

[0224] Off-white solid,yield:94%,mp:182.8–185.3℃. 1H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.15(s,1H),7.72(d,J=8Hz,2H),7.67(d,J= 4Hz,2H),7.53(s,2H),7.50(t,J=4Hz,1H),7.38(d,J=8Hz,2H),6.71(d,J=4Hz,1H),4.85(t ,J=4Hz,1H),4.54(d,J=4Hz,2H),3.39(s,2H),3.31-3.28(m,2H),3.20(s,1H),3.15(s,3H) ,2.95-2.91(m,2H),2.65-2.63(m,2H),2.14(s,6H),1.80-1.63(m,4H),1.40-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.7,159.2,151.4,150.6,147.2,143.9,139.9,134.5,131.9,129 .5,127.2,126.9,121.5,109.4,71.1,62.0,58.3,56.4,52.7,49.0,42.7,31.7,16.6.HRMScalcd.for C 33 H 41 N6O5S + [M+H] + :633.2854, found633.2845.HPLC analysis:retention time=6.40min;peak area,97.93%(λ=254nm).IR:3330,2816,1612,1577,1411,1182cm -1 .

[0225] Example 30: Synthesis of Final Product IId

[0226]

[0227] The operation is the same as Example 27.

[0228] Off-white solid,yield:85%,mp:202.2–204.4℃. 1H NMR (400MHz, DMSO-d6): δ8.63(d,J=4Hz,2H),8.15(s,1H),7.67(d,J=4Hz,2H) ,7.64(d,J=8Hz,2H),7.54(s,2H),7.43(d,J=8Hz,2H),6.71(d,J=4Hz,1H),4.9 0(s,1H),4.57(s,1H),4.53(s,2H),3.54(s,2H),3.21-3.15(m,4H),2.77-2.73 (m,2H),2.66-2.63(m,2H),2.13(s,6H),1.82-1.64(m,6H),1.46-1.36(m,4H). 13 CNMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.4,150.6,147.2,144.7,135.2,134.5,131. 9,129.6,127.7,127.2,121.5,109.4,64.4,61.9,56.4,52.8,49.0,43.6,33.5,31.7,16.6.HRMS calcd.for C 35 H 43 N6O5S + [M+H] + :659.3010, found659.3012.HPLC analysis:retention time=6.50min;peak area,97.06%(λ=254nm).IR:3388,2831,1602,1568,1514,1161cm -1 .

[0229] Example 31: Synthesis of Final Product IIe

[0230]

[0231] The operation is the same as Example 27.

[0232] Off-white solid,yield:90%,mp:218.9–221.1℃. 1H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.15(s,1H),7.76(d,J=8Hz,2H),7.68(d,J=4Hz,2H),7.54(s,2H),7.35(d,J=8Hz,2H),7.18 (s,2H),6.70(d,J=4Hz,1H),4.87(s,1H),4.53(s,2H),3.38(s,3H),2.66-2.63(m,2H),2.14(s,6H),1.80-1.63(m,4H),1.40-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.5,161.6,159.2,151.4,150.6,147.2,143.3,143.3,134 .5,131.9,129.3,127.2,126.1,121.5,109.4,62.0,56.4,52.7,49.0,31.7,16.6.HRMS calcd.forC 30 H 35 N6O4S + [M+H] + :575.2435, found 575.2438.HPLC analysis:retention time=6.03min;peak area,99.62%(λ=254nm).IR:3242,2950,2812,1616,1600,1415,1151cm -1 .

[0233] Example 32: Synthesis of Final Product IIf

[0234]

[0235] The operation is the same as Example 27.

[0236] Off-white solid,yield:87%,mp:134.9–137.0℃. 1H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.25(q,J=4Hz,1H),8.15(s,1 H),7.75(d,J=8Hz,2H),7.67(d,J=4Hz,2H),7.53(s,2H),7.24(d,J=8Hz,2H) ,6.70(d,J=4Hz,1H),4.90(s,1H),4.53(s,2H),3.36(s,3H),2.79(d,J=4Hz ,1H),2.66-2.64(m,2H),2.13(s,6H),1.80-1.63(m,4H),1.40-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ167.1,166.5,161.6,159.2,151.4,150.6,147.2,142.0,134.5,133 .9,131.9,128.9,127.4,127.1,121.5,109.4,62.1,56.4,52.6,49.0,31.6,26.6,16.6.HRMS calcd.for C 32 H 37 N6O3 + [M+H] + :553.2922, found 553.2916.HPLCanalysis:retention time=6.05min;peak area,98.99%(λ=254nm).IR:3427,3327,3225,2940,1617,1413,1152cm -1 .

[0237] Example 33: Synthesis of Final Product IIg

[0238]

[0239] The operation is the same as Example 27.

[0240] Off-white solid,yield:87%,mp:137.1–139.4℃. 1H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.27(t,J=4Hz,1H),8.15(s,1H),7.79(d,J=8Hz,2H),7.67(d,J=4Hz,2H),7.53(s,2H),7.25(d,J=8 Hz,2H),6.69(s,1H),4.90(s,1H),4.66(s,1H),4.53(s,2H),3.56-3.35 (m,7H),2.66(s,2H),2.13(s,6H),1.79-1.62(m,4H),1.40-1.31(m,2H). 13 CNMR (100MHz, DMSO-d6): δ166.8,166.5,161.6,159.2,151.4,150.6,147.2,142.1,134.5,133.9 ,131.9,128.9,127.6,127.1,121.5,109.4,62.1,60.4,56.4,52.6,49.0,42.8,31.6,16.6.HRMS calcd.for C 33 H 39 N6O4 + [M+H] + :583.3027, found 583.3021.HPLC analysis:retention time=5.94min;peak area,98.70%(λ=254nm).IR:3423,3215,2933,1615,1414,1159cm -1 .

[0241] Example 34: Synthesis of Final Product IIh

[0242]

[0243] The operation is the same as Example 27.

[0244] Off-white solid,yield:82%,mp:130.6–132.9℃. 1H NMR (400MHz, DMSO-d6): δ8.64(d,J=4Hz,2H),8.27(t,J=4Hz,1H),8.14(s,1H),7.75(d,J=8Hz,2H),7.67(d,J=4Hz,2H),7.53(s,2H),7.24(d,J=8Hz ,2H),6.68(s,1H),4.89(s,1H),4.52(s,2H),4.38(s,1H),3.48-3.30(m,7 H),2.66-2.64(m,2H),2.13(s,6H),1.79-1.62(m,6H),1.40-1.34(m,2H). 13 C NMR (100MHz, DMSO-d6): δ166.7,166.5,161.6,159.2,151.4,150.6,147.2,142.0,134.5,134.0,13 1.9,128.9,127.5,127.1,121.5,109.4,62.1,59.2,56.4,52.6,49.0,37.1,32.9,31.6,16.6.HRMS calcd.for C 34 H 41 N6O4 + [M+H] + :597.3184, found597.3167.HPLC analysis:retention time=6.10min;peak area,99.61%(λ=254nm).IR:3235,2946,1616,1417,1153cm -1 .

[0245] Example 35: Synthesis of Final Product IIi

[0246]

[0247] The operation is the same as Example 27.

[0248] Off-white solid,yield:84%,mp:143.5–145.7℃. 1H NMR (400MHz, DMSO-d6): δ8.64(s,2H),8.50(t,J=4Hz,1H),8.12(s,1H),7.74(d,J =8Hz,2H),7.70(d,J=4Hz,2H),7.56(s,2H),7.36-7.33(m,1H),7.17-7.13(m,1H), 7.00-6.80(m,1H),5.04(t,J=4Hz,1H),4.49(d,J=4Hz,2H),3.45-3.42(m,7H),3. 25(s,3H),3.21(s,1H),2.72(s,2H),2.11(s,6H),1.95(s,1H),1.69-1.46(m,4H). 13 C NMR (100MHz, DMSO-d6): δ166.6,166.5,161.5,158.9,151.2,150.6,147.1,142.4,134.4,133.5,13 1.8,128.9,127.6,127.3,121.5,109.8,71.0,62.2,58.4,56.4,52.7,48.6,31.8,29.5,16.7.HRMS calcd.for C 34 H 41 N6O4 + [M+H] + :597.3184, found 597.3172.HPLC analysis:retention time=6.28min;peak area,96.94%(λ=254nm).IR:3227,2943,1616cm -1 .

[0249] Example 36: Synthesis of Final Product IIj

[0250]

[0251] The operation is the same as Example 27.

[0252] Off-white solid,yield:93%,mp:145.5–147.6℃. 1H NMR (400MHz, DMSO-d6): δ8.63(d,J=4Hz,2H),8.15(s,1H),7.75(s,3H),7.66(d,J=4Hz,2H),7.53(s,2H),7.32(s,2H),7.15( s,1H),6.68(s,1H),4.90(s,1H),4.53(s,2H),3.37(s,3H),2.66(s,2H),2.13(s,6H),1.80-1.63(m,4H),1.41-1.32(m,2H). 13 C NMR (100MHz, DMSO-d6): δ168.4,166.5,161.5,158.8,151.3,150.7,147.1,139.1,134.7, 134.4,132.1,131.8,128.4,127.2,126.5,121.5,62.4,56.4,52.7,48.5,31.6,16.7.HRMS calcd.for C 31 H 35 N6O3 + [M+H] + :539.2765, found 539.2755.HPLC analysis: retention time=5.93min;peak area,99.68%(λ=254nm).IR:3250,2930,1615,1602,1414cm -1 .

[0253] Example 37: Synthesis of Final Product IIk

[0254]

[0255] The operation is the same as Example 27.

[0256] Off-white solid,yield:92%,mp:171.7–173.8℃. 1 H NMR (400 MHz, DMSO-d d6): δ8.64(d,J=4Hz,2H),8.15(s,1H),7.80(d,J=8Hz,2H),7.73(s,1H),7.67(d,J=4Hz,2H),7.53(s,2H),7.24(d,J=8Hz,2H),7. 12(s,1H),6.70(s,1H),4.89(s,1H),4.53(s,2H),3.36(s,3H),2.65(s,2H),2.13(s,6H),1.79-1.62(m,4H),1.40-1.31(m,2H). 13 C NMR (100MHz, DMSO-d6): δ168.4,166.5,161.6,159.2,151.4,150.6,147.2,142.4,134.5, 133.6,131.9,128.8,127.9,127.1,121.5,109.4,62.2,56.4,52.7,49.0,31.6,16.6.HRMS calcd.forC 31 H 35 N6O3 + [M+H] + :539.2765, found 539.2766.HPLC analysis:retention time=5.99min;peak area,99.87%(λ=254nm).IR:3513,3346,3255,29571616,1412,1168cm -1 .

[0257] Example 38: Anti-HIV Biological Activity Test

[0258] The Rega Institute of Drugs at the Catholic University of Leuven in Belgium evaluated the anti-HIV activity at the in vitro cellular level, mainly examining two aspects: one is the inhibitory effect on HIV infection of MT-4 cells, and the other is the toxic effect on cells. The specific steps of the experiment are as follows: the compound to be tested is added to the MT-4 cell culture medium, and the cells are attacked by HIV virus at different time points. The MTT method is used to evaluate the protective effect of the drug on HIV-induced cell damage, and the minimum concentration (EC 50 ). While conducting the anti-HIV activity experiment, a cytotoxicity test was also conducted. The MTT method was also used to determine the minimum concentration (CC) that damages 50% of cells that are not infected with HIV. 50 ) Calculate the selectivity index (SI), the formula is SI = EC 50 / CC 50, to evaluate the safety and efficacy of drugs. Through this method, researchers can comprehensively evaluate the anti-HIV activity and cytotoxicity of compounds, providing important reference for drug development.

[0259] Materials and methods: The anti-HIV activity of each compound was monitored by the efficiency of the drug's inhibitory effect on HIV-induced cytopathic effects in cells. MT-4 cells were used for cell culture. The virus strains used were: HIV-1 virus strain ⅢB and common clinical mutant strains and HIV-2 virus strain ROD.

[0260] The experimental procedure is as follows: First, the compound to be tested is dissolved in DMSO or water and diluted with phosphate buffered saline (PBS). ^ 5 MT-4 cells were pre-cultured with 100 μL of compound solutions of different concentrations at 37°C for 1 hour. Then, 100 μL of appropriately diluted virus solution was added to the cells and cultured at 37°C for another hour. After the culture, the cells were washed three times with PBS and resuspended in culture medium with or without the compound. After that, the cells were placed in a 5% CO2 environment and cultured at 37°C for 7 days, during which the culture medium with or without the compound was replaced on the third day after infection. Each culture condition was repeated twice. The virus-induced cytopathic effect was observed every day using an inverted optical microscope. Usually, the virus dilution used in the experiment will cause cell pathology on the fifth day after infection. Drug inhibitory concentration (EC 50 ) refers to the drug concentration that reduces virus-induced cytopathic effects by 50% without directly damaging cells. It is worth noting that when a compound is poorly water-soluble and needs to be dissolved in DMSO, the final concentration of DMSO should be less than 10% (less than 2% in MT-4 cell culture medium) because DMSO may affect the antiviral activity of the compound. Therefore, for solutions containing the same DMSO concentration, a parallel comparison of antiviral activity should also be performed with a blank control group. In addition, the final concentration of DMSO (1 / 1000) is much lower than the concentration required for HIV-1 to replicate in T cells.

[0261] The present invention uses the marketed anti-HIV-1 drugs Nevirapine (NVP), Efavirenz (EFV) and Etravirine (ETR) as reference substances, and the inhibitory activities of some target compounds on HIV-1 wild strains and Y188L, double mutant strains (F227L+V106A, RES056) and cytotoxicity to MT-4 cells are shown in Table 1.

[0262] Table 1: Inhibitory activity against HIV-1 wild-type, Y188L and double mutants (F227L+V106A, RES056) and cytotoxicity against MT-4 cells

[0263]

[0264]

[0265]

[0266] Table 2: Inhibitory activity of some compounds against other common single mutant strains of HIV-1

[0267]

[0268] The experimental results show that most of the target products I and II of the examples have strong anti-HIV-1 activity, can significantly inhibit viral replication in MT-4 cells infected with HIV-1, and have low cytotoxicity and high selectivity index.

[0269] It should be noted that the technical parameters and raw material components not described in detail in the above embodiments can still achieve the same or similar technical effects as the above embodiments when they are changed within the parameter range listed in the present invention. Therefore, the changes and modifications made to the embodiments, or the equivalent transformations made using the contents disclosed in the present specification, are all included in the patent protection scope of the present invention.

Claims

1. A non-nucleoside reverse transcriptase inhibitor of HIV-1, characterized in that: It is a piperidine-hydroxymethyl-diaryl pyrimidine derivative, and its structural formula is shown in (Ⅰ) or (ⅠⅠ): Wherein, R is selected from substituted or unsubstituted benzene ring.

2. The HIV-1 non-nucleoside reverse transcriptase inhibitor according to claim 1, characterized in that: Also included are pharmaceutically acceptable salts, stereochemical isomers, hydrates or solvates of piperidine-hydroxymethyl-diarylpyrimidine derivatives.

3. The HIV-1 non-nucleoside reverse transcriptase inhibitor according to claim 2, characterized in that: The pharmaceutically acceptable salt is hydrochloride, hydrobromide, formate, methanesulfonate, trifluoromethanesulfonate, sulfate, phosphate, acetate, p-toluenesulfonate, tartrate, citrate, succinate, maleate, fumarate or malate.

4. The method for preparing the HIV-1 non-nucleoside reverse transcriptase inhibitor according to claim 1, characterized in that: The specific steps are as follows: Step 1: Using 2-thioether-4-chloro-5-ethylpyrimidine as a raw material, react with 4-bromo-2,6-dimethylphenol to obtain compound III, the reaction formula of which is as follows: Specifically, a base is added to a solution containing 2-methylsulfide-4-chloro-5-ethylpyrimidine and 4,6-dimethylphenol; the mixture is stirred at a certain temperature for reaction; after the reaction is completed, the mixture is added dropwise to water, the filtered solid is dissolved with dichloromethane, washed with water and a saturated sodium chloride solution, dried with anhydrous sodium sulfate, and concentrated to obtain a residue; the residue is then added dropwise to methanol to precipitate compound III as a yellow solid, which is dried in a vacuum oven and used for the next reaction; wherein: The solvent is selected from N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene, and 1,2-dichloroethane; The alkali is selected from sodium carbonate, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, potassium phosphate; The molar ratio of the compound 2-thioether-4-chloro-5-ethylpyrimidine, 4-bromo-2,6-dimethylphenol, and base is 1:(1.0-3.0):(1-10); The reaction temperature is 0 to 180°C; The reaction time is 2 to 24 hours; Step 2: Use an oxidant to oxidize the methylthio group of compound III to sulfone to obtain compound IV, the reaction formula of which is as follows: Specifically, an oxidant is added to a solution of compound III and stirred at a certain temperature for reaction; after the reaction is completed, a reducing alkaline solution is added to the reaction solution; the separated organic phase is washed with water and a saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to obtain a crude product IV as a light yellow solid, which is directly used in the next reaction without purification; wherein: The solvent is selected from dichloromethane, ethyl acetate, methyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene; The oxidant is selected from meta-chloroperbenzoic acid, hydrogen peroxide, peracetic acid, potassium monopersulfate composite salt, potassium persulfate; The molar ratio of the compound III to the oxidant is 1:(2.0-20.0); The reducing alkaline solution is selected from sodium thiosulfate, sodium sulfite, and sodium bisulfite; The reaction temperature is -20 to 100°C; The reaction time is 0.5 to 24 hours; Step 3: The methylthio group of compound IV undergoes a nucleophilic substitution reaction with 4-amino-1-Boc piperidine to obtain compound V, the reaction formula of which is as follows: Specifically, a base is added to a solution of compound IV and 4-amino-1-tert-butyloxycarbonyl-piperidine, and the mixture is stirred at a certain temperature to react; after the reaction is completed, the reaction mixture is filtered; the filtered off-white solid is placed in water and stirred, filtered, and the obtained solid is rinsed with water; the obtained solid is vacuum dried to obtain compound V, which is an off-white solid; wherein: The solvent is selected from acetonitrile, dichloromethane, ethyl acetate, methyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene; The base is selected from sodium carbonate, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, potassium phosphate; The molar ratio of the compound IV to 4-amino-1-tert-butyloxycarbonyl-piperidine is 1:(1.0-10.0); The reaction temperature is 0 to 180°C; The reaction time is 0.5 to 24 hours; Step 4: Reducing the ethyl ester group of compound V to a hydroxyl group to obtain compound VI, the reaction formula of which is as follows: Specifically, at a certain temperature, a reducing agent is added to a solution of compound V, and the mixture is stirred for reaction; after the reaction is completed, an alkaline solution is added to quench the reaction; dichloromethane is then added for extraction; the organic phase is dried over anhydrous sodium sulfate and concentrated to obtain a crude product; ethyl acetate and petroleum ether are used as mobile phases, and the compound VI is purified by silica gel column chromatography to obtain a white solid; wherein: The solvent is selected from tetrahydrofuran, dichloromethane, dioxane, toluene, and 1,2-dichloroethane; The reducing agent is selected from lithium aluminum tetrahydride, diisobutylaluminum hydride, sodium borohydride, and tributyltin hydride; The molar ratio of the compound V to the reducing agent is 1:(1.0-20.0); The reaction temperature is -78 to 100°C; The reaction time is 0.5 to 16 hours; Step 5: The bromine of compound VI undergoes Suzuki coupling reaction with 4-cyanophenylboronic acid or pyridine-4-boronic acid to obtain compound VIIa or VIIb, and the reaction formula is as follows: Specifically, compound VI, 4-cyanophenylboronic acid or pyridine-4-boronic acid and a base are added to a solvent, a palladium catalyst is added, the mixture is degassed and replaced with nitrogen three times, and the reaction is stirred at a certain temperature; after the reaction is completed, the reaction solution is concentrated; the obtained residue is purified by column chromatography, eluted with ethyl acetate and petroleum ether; the eluate is concentrated and vacuum dried to obtain compound VIIa or VIIb, which is an off-white solid; wherein: The solvent is selected from acetonitrile, dichloromethane, ethyl acetate, methyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, toluene, 1,2-dichloroethane; The base is selected from sodium carbonate, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, potassium phosphate; The molar ratio of the compound VI, 4-cyanophenylboronic acid or pyridine-4-boronic acid, palladium catalyst and base is 1:(1.0-10.0);(0.01%-1.0);(2.0-10.0); The reaction temperature is 50-180°C; The reaction time is 2 to 48 hours; Step 6: Remove the Boc protecting group of compound VIIa or VIIb with acid to obtain compound VIIa or VIIb; the reaction formula is as follows: Specifically, an acid is added to a solution of compound VIIa or VIIb, and the reaction is stirred at a certain temperature; after the reaction is completed, the reaction solution is added dropwise to an alkaline solution to quench, and then extracted with dichloromethane; the organic phase is washed with water and a saturated sodium chloride solution, dried with anhydrous sodium sulfate and concentrated, and finally filtered to obtain compound VIIIa or VIIIb, which is an off-white solid; wherein: The solvent is selected from dichloromethane, dioxane, acetone, ethanol, methanol, isopropanol, tert-butanol, acetonitrile, tetrahydrofuran, toluene, and 1,2-dichloroethane; The acid is selected from hydrochloric acid, hydrobromic acid, trifluoroacetic acid, sulfuric acid, and glacial acetic acid; The reaction temperature is -50 to 100°C; The reaction time is 0.5 to 48 hours; Step 7: The piperidine nitrogen atom of compound VIIIa or VIIIb reacts with a substituted benzyl halide to obtain compound I or II; the general reaction formula is as follows: Specifically, at a certain temperature, a base is added to a solvent containing compound VIIIa or VIIIb and a substituted benzyl halide, and the mixture is stirred at a certain temperature for reaction; after the reaction is completed, the reaction solution is poured into water and filtered to obtain a solid crude product; using dichloromethane and methanol as mobile phases, the final products I and II are purified by column chromatography, which are yellow or off-white solids; wherein: The solvent is selected from N,N-dimethylformamide, dimethyl sulfoxide, dioxane, acetone, acetonitrile, tetrahydrofuran, N,N-dimethylacetamide, ethanol, isopropanol, n-butanol, isobutanol; The base is selected from sodium carbonate, potassium carbonate, sodium hydroxide, potassium phosphate; The substituted benzyl halide RCH2X is one of the following: chloro, bromo, iodo; The molar ratio of the raw material compound VIIIa or compound VIIIb, substituted benzyl halide and base is 1:(1.0-2.0):(1-5); The reaction temperature is 0 to 180°C; The reaction time is 2 to 24 hours.

5. A pharmaceutical composition, characterized in that The invention comprises an effective dose of the HIV-1 non-nucleoside reverse transcriptase inhibitor as claimed in any one of claims 1 to 3 and a pharmaceutical carrier.

6. Use of the HIV-1 non-nucleoside reverse transcriptase inhibitor according to any one of claims 1 to 3 in the preparation of a drug for preventing and treating AIDS.

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