Phenylsulfonamide piperidinone glycine derivatives, processes for their preparation and use

Abstract

The application provides a benzene sulfonamide piperazine ketone glycine derivative and a preparation method and application thereof. The derivative has a structure shown in the following general formula I. The application also relates to a preparation method of a compound containing the structure of formula I and application of the compound as an HIV-1 / 2 capsid protein regulator in preparation of an anti-AIDS drug.

Description

Technical Field

[0001] This invention belongs to the field of organic compound synthesis and pharmaceutical application technology, specifically relating to a benzenesulfonamide piperazine glycine derivative, its preparation method, and its use as an anti-HIV drug. Background Technology

[0002] AIDS (Acquired Immunodeficiency Syndrome) is a chronic infectious disease caused by the Human Immunodeficiency Virus (HIV). HIV is an RNA retrovirus, with two subtypes: HIV-1 and HIV-2. HIV-1 is the currently prevalent pathogen. HIV-2 is mainly prevalent in West Africa, but with globalization and increasingly frequent personnel exchanges, the risk of HIV-2 infection is constantly increasing, which should be given sufficient attention. Currently, anti-HIV drugs remain an effective weapon in the fight against AIDS. More than 30 chemical entities targeting key stages of the HIV-1 life cycle have been approved for marketing for combination therapy. However, drug resistance caused by rapid HIV mutations has become the biggest challenge facing existing anti-AIDS drugs, forcing researchers to develop anti-AIDS drugs with new targets, new mechanisms, and new structures. The HIV-1 capsid protein is a structural protein essential for the formation of morphologically mature, infectious viral particles, internally encapsulating nucleic acids and enzymes crucial for viral infection. It plays a crucial role in both the early (uncoating, reverse transcription, nucleus entry, etc.) and late (assembly, maturation) stages of viral replication, and is a new target for anti-AIDS drug research.

[0003] Proteolysis Targeting Chimeria (PROTAC) is a technology that utilizes the intracellular ubiquitin-proteasome degradation system to target and degrade target proteins. PROTAC is a bifunctional molecule composed of a target protein ligand, an intermediate linker chain, and an E3 ubiquitin ligase ligand. It can simultaneously recognize the target protein and the E3 ligase, forming a triplet complex with them spatially, subsequently mediating the ubiquitination and degradation of the target protein. Due to its multiple advantages, PROTAC technology has been widely applied to various targets. Therefore, designing degradative agents using the PROTAC strategy against the multifunctional HIV capsid protein, and interfering with its normal function through a novel mechanism of action, holds promise for unlocking greater antiviral potential.

[0004] Our research group previously discovered a novel HIV capsid protein regulator, I-3L, using PF74 (reported by Pfizer) as a lead compound and through structural modification. However, its antiviral activity still needs improvement. Based on the structural biology information of the I-3L-HIV-1 capsid protein cocrystal, this invention utilizes the PROTAC strategy, employing the terminal NH2 exposed in the solvent opening region as the linking site, and connecting E3 ubiquitin ligands with linkers of varying lengths to design and synthesize a novel benzylamide-piperazinone glycine derivative. This derivative holds promise for improving the low efficacy and severe drug resistance issues of existing HIV capsid protein regulators. No similar compounds have been reported in the prior art.

[0005] Summary of the Invention

[0006] This invention provides a benzylsulfonamide piperazine glycine derivative, its preparation method and application, and also provides the activity screening results of the above compound against HIV-1 / 2 and its pharmaceutical application.

[0007] The technical solution of the present invention is as follows:

[0008] 1. Benzenesulfonamide piperazine glycine derivative

[0009] The benzenesulfonamide piperazine glycine derivative involved in this invention has the structure shown in general formula I:

[0010]

[0011] Linker is a linking group, selected from aliphatic amino acid chains with 3-20 atoms in the main chain;

[0012] R is a H atom, pomalidomide, 1-methylpomalidomide, lenalidomide, 1-methyllenalidomide, thalidomide, or 1-methylthalidomide.

[0013] According to a preferred embodiment of the present invention, the Linker is an aliphatic amino acid chain with a main chain number of 4-10 atoms; more preferably, it is a compound with the structure shown below:

[0014]

[0015] According to a further preferred embodiment of the present invention, the benzenesulfonamide piperazine ketone glycine derivative is a compound having one of the following structures:

[0016]

[0017] 2. Preparation method of benzenesulfonamide piperazine ketone glycine derivative

[0018] This invention also provides a method for preparing the benzenesulfonamide piperazine glycine derivative, the synthesis steps of which are as follows: 3-fluorophthalic anhydride I-1 is used as a raw material and 3-amino-2,6-piperidindione hydrochloride and sodium acetate are refluxed at 120°C for 10 h in acetic acid solvent to obtain intermediate I-2; I-2 is dissolved in N,N-dimethylformamide solution, potassium carbonate is used as a base, iodomethane is added under stirring, and the reaction is carried out at room temperature for 24 h to obtain intermediate I-3; N-Boc-L-phenylalanine (1) is used as a starting material and N-methyl-4-aminoanisole undergoes an amide condensation reaction in dichloromethane to obtain intermediate 2; 2 is used as a reaction solvent and the Boc group is removed under the action of trifluoroacetic acid to obtain intermediate 3; then intermediate 3 undergoes an amide condensation reaction with bromoacetic acid in dichloromethane to obtain intermediate 4; 4 reacts with 1-Boc-3-piperazinone in N,N-dimethylformamide as a reaction solvent and undergoes an S-reaction reaction under the action of cesium carbonate. N 2. A nucleophilic substitution reaction yields intermediate 5; subsequently, 5 is deactivated with the Boc group in trifluoroacetic acid using dichloromethane as the reaction solvent to obtain intermediate 6; then, 6 is reacted with p-nitrobenzenesulfonyl chloride in dichloromethane as the reaction solvent, followed by an acylation reaction in triethylamine to obtain intermediate 7; 7 is then subjected to a hydrogenation reduction reaction in methanol and dichloromethane using 10% palladium on carbon as the reaction solvent to obtain the key intermediate 8; N-tert-butoxycarbonyl carboxylic acid fragments of different lengths and 2-(7-azabenzotriazole)-N,N,N',N' -Tetramethylurea hexafluorophosphate was reacted in an ice-water bath for 30 min, and intermediate 8 and N,N-diisopropylethylamine were added and stirred overnight at room temperature to obtain intermediate 9(ae); 9(ae) was reacted with dichloromethane as a reaction solvent and the Boc group was removed in the presence of trifluoroacetic acid to obtain the target compound 10(ae); 10(ae) was reacted with I-2 or I-3 in N,N-dimethylformamide solution with N,N-diisopropylethylamine as a base and refluxed at 90 °C for 10 h to obtain the target compound 11(ae) ​​or 11(fj);

[0019] The synthesis route is as follows:

[0020]

[0021] Reagents and conditions: (i) 3-amino-2,6-piperidinidone hydrochloride, acetic acid, sodium acetate, 120℃, 10h; (ii) potassium carbonate, iodomethane, N,N-dimethylformamide, room temperature, 24h; (iii) N-methyl-4-aminoanisole, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, 0℃→room temperature; (iv) trifluoroacetic acid, dichloromethane, 0℃→room temperature; (v) bromoacetic acid, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, 0℃→room temperature; (vi) 1-Boc-3-piperazine Ketone, cesium carbonate, N,N-dimethylformamide, 45°C; (vii) trifluoroacetic acid, dichloromethane, 0°C → room temperature; (viii) p-nitrobenzenesulfonyl chloride, triethylamine, dichloromethane, 0°C → room temperature; (ix) H2, 10% Pd·C, dichloromethane / methanol, room temperature; (x) N-tert-butoxycarbonylcarboxylic acid fragments of different lengths, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, 0°C, 30 min, room temperature; (xi) trifluoroacetic acid, dichloromethane, 0°C → room temperature; (xii) I-2 or I-3, N,N-dimethylformamide, N,N-diisopropylethylamine, 90°C, 10 h.

[0022] The N-tert-butoxycarbonyl carboxylic acid fragments of different lengths include: N-tert-butoxycarbonyl-4-aminobutyric acid, N-tert-butoxycarbonyl-6-aminohexanoic acid, N-tert-butoxycarbonyl-7-aminoheptanoic acid, N-tert-butoxycarbonyl-8-aminooctanoic acid, and N-tert-butoxycarbonyl-10-aminodecanoic acid.

[0023] The room temperature described in this invention is 20-30℃.

[0024] According to a preferred embodiment of the present invention, the preparation method of the benzenesulfonamide piperazine ketone glycine derivative of the present invention comprises the following specific steps:

[0025] (1) Dissolve 3-fluorophthalic anhydride (I-1), 3-amino-2,6-piperidindione hydrochloride and sodium acetate in acetic acid and reflux at 120°C for 10 h. After the reaction is complete, remove the solvent under reduced pressure, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and obtain I-2 after separation and purification by silica gel column chromatography.

[0026] (2) I-2 and potassium carbonate were added to N,N-dimethylformamide, and then iodomethane was added dropwise. The mixture was stirred at room temperature for 24 hours. After the reaction was completed, the solvent was removed by vacuum distillation, the mixture was extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain I-3.

[0027] (3) Boc-L-phenylalanine (1) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate were added to dichloromethane and stirred in an ice bath for 30 min. Then N,N-diisopropylethylamine and N-methyl-4-aminoanisole were added to the reaction solution, the ice bath was removed and the mixture was brought to room temperature, and TLC was used for monitoring. After the reaction was completed, the solvent was removed by vacuum distillation and the mixture was extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain intermediate 2.

[0028] (4) Add intermediate 2 obtained in the previous step to dichloromethane. Under ice bath conditions and stirring, slowly add trifluoroacetic acid to this solution. After the addition is complete, remove the ice bath and bring to room temperature. Monitor the reaction by TLC. After the reaction is complete, remove the solvent under reduced pressure. Then add saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7. Then add dichloromethane solution for extraction. Dry the organic phase with anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain intermediate 3.

[0029] (5) Bromoacetic acid and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate were added to dichloromethane and stirred in an ice bath for 30 min. Then, intermediate 3 and N,N-diisopropylethylamine were added to this solution, the ice bath was removed, and the mixture was brought to room temperature. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain intermediate 4.

[0030] (6) Intermediate 4, 1-Boc-3-piperazinone and cesium carbonate were added to N,N-dimethylformamide and stirred at 45°C for 12 h from room temperature. After the reaction was completed, an appropriate amount of saturated sodium chloride solution was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain intermediate 5.

[0031] (7) Add intermediate 5 to dichloromethane and slowly add trifluoroacetic acid dropwise under stirring in an ice bath. After the addition is complete, remove the ice bath and bring the mixture to room temperature. Monitor the reaction by TLC. After the reaction is complete, remove the solvent under reduced pressure and then add saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7. Then add dichloromethane solution for extraction. Dry the organic phase with anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain intermediate 6.

[0032] (8) Add intermediate 6 and triethylamine to dichloromethane, and slowly add p-nitrobenzenesulfonyl chloride under stirring in an ice bath. Then remove the ice bath and bring to room temperature. Monitor by TLC. After the reaction is complete, add saturated sodium chloride solution, extract with dichloromethane, dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and obtain intermediate 7 by silica gel column chromatography.

[0033] (9) Dissolve intermediate 7 in methanol and dichloromethane, add 10% palladium on carbon, replace with hydrogen three times, stir overnight at room temperature under hydrogen balloon protection; after the reaction is complete, filter with diatomaceous earth, evaporate the filtrate to dryness to obtain intermediate 8;

[0034] (10) N-tert-butoxycarbonylcarboxylic acid fragments of different lengths and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate were added to dichloromethane and stirred in an ice bath for 30 min. Then, intermediate 8 and N,N-diisopropylethylamine were added to the reaction solution, the ice bath was removed and the mixture was brought to room temperature, and TLC was used for monitoring. After the reaction was completed, the solvent was removed by vacuum distillation and the mixture was extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain intermediate 9(ae).

[0035] (11) Intermediate 9(ae) was dissolved in dichloromethane, and trifluoroacetic acid was slowly added dropwise under stirring in an ice bath. After the addition was complete, the ice bath was removed and the mixture was brought to room temperature. The reaction was monitored by TLC. After the reaction was completed, the solvent was removed by vacuum distillation, and then saturated sodium bicarbonate solution was added to adjust the pH of the reaction solution to 7. Then, dichloromethane solution was added for extraction. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain the target compound 10(ae).

[0036] (12) Dissolve 10(ae) and I-2 or I-3 in N,N-dimethylformamide, add N,N-diisopropylethylamine, and react at 90°C for 10 h. After the reaction is complete, remove the solvent under reduced pressure, extract with water and ethyl acetate, dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography to obtain the target compound 11(aj).

[0037] 3. Applications of benzenesulfonamide piperazine glycine derivatives

[0038] This invention discloses the screening results of the anti-HIV-1 / 2 activity of the benzylamide piperazine glycine derivative and its first application in the preparation of HIV-1 / 2 drugs. Experiments demonstrate that the benzylamide piperazine glycine derivative of this invention can be used as an HIV-1 / 2 capsid protein regulator in the preparation of anti-AIDS drugs.

[0039] Anti-HIV-1 / 2 activity and cytotoxicity assays of the target compound

[0040] The anti-HIV-1 / 2 activity and toxicity of a benzylamide piperazine glycine derivative synthesized according to the above method were tested at the cellular level. Their anti-HIV-1 and HIV-2 activity and toxicity data are listed in Table 1, with the capsid protein regulators PF74 and I-3L reported in the literature as positive controls.

[0041] The newly synthesized benzylsulfonamide piperazine glycine derivatives of this invention exhibit significant anti-HIV activity. All compounds (except 10b) showed anti-HIV-1 activity at low micromolar concentration levels, EC50. 50 The values ​​ranged from 0.35 to 4.39 μM. Among them, 11h (EC) 50 =0.35±0.02μM) and I-3L (EC 50 =0.24±0.12μM) has comparable activity to PF74 (EC) 50 =2.47±1.36μM) 7 times; in addition, 10d (EC 50 =0.79±0.15μM), 11b (EC 50 =0.97±0.17μM), 11c(EC) 50 =0.71±0.19μM), 11d(EC) 50 (0.88±0.33 μM) also showed good activity, with most compounds in this series exhibiting better activity than PF74. Furthermore, this series of compounds possesses superior anti-HIV-2 activity, EC50... 50 The values ​​ranged from 0.02 to 1.54 μM, among which 10a (EC) 50(IIIB) =3.29±1.34μM, EC 50(ROD) =0.04±0.02μM, Ratio (IIIB / ROD) =82.25) is a selective regulator of HIV-2 capsid protein. Therefore, the newly synthesized benzylsulfonamide piperazine glycine derivative of this invention is worthy of further research.

[0042] Effect of the target compound on HIV-1 capsid protein content

[0043] The results of the effect of representative compounds on HIV-1 capsid protein content are as follows: Figure 1 As shown. Figure 1 A represents the result obtained without the addition of the proteasome inhibitor MG-132. Figure 1 B represents the result obtained after adding MG-132. Test results show that compound 11h can effectively reduce the content of HIV-1 capsid protein and preliminarily demonstrates that it may be degraded via the proteasome pathway. Therefore, this invention provides a new direction for the development of anti-HIV drugs.

[0044] The benzylsulfonamide piperazine glycine derivative of the present invention is a novel HIV capsid protein regulator that can be used as an anti-HIV-1 / 2 drug.

[0045] This invention utilizes a constructed HIV-1 capsid protein degradation screening model and, through synthesis optimization, discovered that the synthesized representative compounds can effectively reduce the expression level of HIV-1 capsid protein in vitro, providing a potential treatment method for treating AIDS through HIV capsid protein.

[0046] An anti-HIV-1 / 2 pharmaceutical composition comprising the benzylsulfonamide piperazine glycine derivative of the present invention and one or more pharmaceutically acceptable carriers or excipients.

[0047] This invention discloses a benzylsulfonamide piperazine glycine derivative, its preparation method, anti-HIV activity screening results, HIV-1 capsid protein degradation results, and its first application as a preparation of anti-HIV-1 / 2 drugs. Attached Figure Description

[0048] Figure 1 This is a graph showing the effect of a representative compound on the content of HIV-1 capsid protein. Detailed Implementation

[0049] The following examples help to understand the present invention, but do not limit the scope of the invention. All percentages are mass percentages.

[0050] Example 1: Preparation of Intermediate I-2

[0051] 3-Fluorophthalic anhydride (2.00 g, 12.04 mmol, 1.0 eq.), 3-amino-2,6-piperidinidone hydrochloride (2.59 g, 12.04 mmol, 1.0 eq.), and sodium acetate (1.18 g, 14.4 mmol, 1.2 eq.) were dissolved in 40 mL of acetic acid and refluxed at 120 °C for 10 h. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was extracted with water (40 mL) and ethyl acetate (40 mL × 3 times). The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under vacuum. The product was separated by silica gel column chromatography to obtain intermediate I-2 2.50 g. White solid product, yield 75%; spectral data: 1H NMR(400MHz,DMSO-d6)δ11.15(s,1H,NH),7.95(q,J=7.4Hz,1H,Ph-H),7.83–7.66(m,2H,Ph-H),5.16(dd,J =12.8,5.1Hz,1H,CH),2.97–2.82(m,1H,CH2),2.68–2.52(m,1H,CH2),2.18–1.96(m,2H,CH2).ESI-MS:m / z 275.06[MH] - C 13 H9FN2O4[276.05].

[0052] Example 2: Preparation of intermediate I-3

[0053] I-2 (2.50 g, 9.05 mmol, 1.0 eq.) and potassium carbonate (1.25 g, 9.05 mmol, 1.0 eq.) were dissolved in 15 mL of N,N-dimethylformamide. Iodimethane (0.56 mL, 9.05 mmol, 1.0 eq.) was added dropwise under stirring at room temperature for 24 h. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was extracted with water (30 mL) and dichloromethane (30 mL × 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography to obtain intermediate I-3 2.00 g. White solid product, yield 76%; spectral data: 1 H NMR(400MHz,DMSO-d6)δ7.96(tdt,J=7.4,4.7,2.3Hz,1H,Ph-H),7.83–7.70(m,2H,Ph-H),5.30–5.12(m,1H,CH),3.08– 2.85(m,3H,CH3),2.85–2.73(m,1H,CH2),2.66–2.51(m,1H,CH2),2.08(tdd,J=12.7,7.5,4.2Hz,1H,CH2).ESI-MS:m / z 289.25[MH] - C 14 H 11 FN2O4[290.07].

[0054] Example 3: Preparation of Intermediate 2

[0055] The starting materials Boc-L-phenylalanine (1) (6.96 g, 26.24 mmol, 1.2 eq.) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (12.47 g, 32.81 mmol, 1.5 eq.) were added to 30 mL of dichloromethane and stirred in an ice bath for 30 min; then N,N-diisopropylethylamine (7.23 mL, 43.74 mmol) was added. 2.0 eq.) and N-methyl-4-aminoanisole (3.00 g, 21.87 mmol, 1.0 eq.) were added, and the mixture was stirred at room temperature after removing the ice bath. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was extracted with water (40 mL) and dichloromethane (40 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography to obtain 27.20 g of the intermediate, a yellow oily substance, with a yield of 86%. Spectroscopic data: 1 H NMR(400MHz,DMSO-d6)δ7.22(d,J=8.3Hz,2H,Ph-H),7.20–7.11(m,3H,Ph-H),7.09 (d,J=8.2Hz,1H,NH),7.03(d,J=8.6Hz,2H,Ph-H),6.79(d,J=7.3Hz,2H,Ph-H),4.2 7–4.06(m,1H,CH),3.81(s,3H,OCH3),3.13(s,3H,NCH3),2.75(dd,J=13.4,3.8Hz, 1H,PhCH),2.61(dd,J=13.3,10.3Hz,1H,PhCH),1.30(s,9H,C(CH3)3).ESI-MS:m / z 385.4 [M+H] + C 22 H 28 N2O4[384.5].

[0056] Example 4: Preparation of Intermediate 3

[0057] Intermediate 2 (7.20 g, 18.73 mmol) was added to 40 mL of dichloromethane. Trifluoroacetic acid (10 mL) was slowly added dropwise to this solution with stirring in an ice bath. After the addition was complete, the ice bath was removed and the solution was brought to room temperature. The reaction was monitored by TLC. After the reaction was complete, the pH of the reaction solution was adjusted to 7 with saturated sodium bicarbonate solution. Water (40 mL) and dichloromethane (40 mL × 3) were added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 34.50 g of intermediate as a yellow oil, with a yield of 84%. Spectroscopic data: 1H NMR(400MHz,DMSO-d6)δ7.29–7.13(m,3H,Ph-H),7.03–6.75(m,6H,Ph-H),3.77(s,3H,OCH3),3.44–3.35(m,1H,CH),3. 06(s,3H,NCH3),2.75(dd,J=12.8,6.7Hz,1H,PhCH),2.45(dd,J=12.9,7.1Hz,1H,PhCH),1.87(s,2H,NH2).ESI-MS:m / z 285.05[M+H] + C 17 H 20 N2O2 [284.36].

[0058] Example 5: Preparation of Intermediate 4

[0059] Bromoacetic acid (2.64 g, 19.00 mmol, 1.2 eq.) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (9.03 g, 23.75 mmol, 1.5 eq.) were added to 30 mL of dichloromethane and stirred in an ice bath for 30 min. Then, intermediate 3 (4.50 g, 15.83 mmol, 1.0 eq.) and N,N-diisopropylethylamine (5.23 mL, 31.66 mmol, 2.0 eq.) were added to this solution. After removing the ice bath, the mixture was stirred at room temperature and monitored by TLC. After the reaction was complete, the solvent was removed under reduced pressure, and the intermediate was separated by silica gel column chromatography to obtain 44.36 g of the intermediate as a white oil, with a yield of 68%. Spectroscopic data: ESI-MS: m / z 405.04 [M+H] + ; 407.12[M+H+2] + C 19 H 21 BrN2O3[404.07].

[0060] Example 6: Preparation of Intermediate 5

[0061] Intermediate 4 (4.30 g, 10.61 mmol, 1.0 eq.), 1-Boc-3-piperazinone (2.55 g, 12.73 mmol, 1.2 eq.), and cesium carbonate (6.91 g, 21.22 mmol, 2.0 eq.) were added to 15 mL of N,N-dimethylformamide, and the mixture was stirred at 40 °C for 12 h after the temperature was increased from room temperature. After the reaction was completed, the solvent was removed by vacuum distillation, and the mixture was extracted with water (30 mL) and ethyl acetate (30 mL × 3 times). The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under vacuum. The intermediate was separated by silica gel column chromatography to obtain 53.62 g of intermediate as a white solid, with a yield of 65%. Spectroscopic data: ESI-MS: m / z 523.09 [MH]-; C28 H 36 N4O6[524.62].

[0062] Example 7: Preparation of Intermediate 6

[0063] Intermediate 5 (3.60 g, 6.86 mmol) was added to 32 mL of dichloromethane. Trifluoroacetic acid (8 mL) was slowly added dropwise with stirring in an ice bath. After the addition was complete, the ice bath was removed and the mixture was brought to room temperature. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed under reduced pressure, and the pH of the reaction solution was adjusted to 7 by adding saturated sodium bicarbonate solution. Then, dichloromethane (30 mL × 3) was added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain 62.18 g of intermediate as a yellow oil, with a yield of 75%. Spectroscopic data: ESI-MS: m / z 425.4 [M+H] + C 23 H 28 N4O4[424.5].

[0064] Example 8: Preparation of Intermediate 7

[0065] Intermediate 6 (2.00 g, 4.71 mmol, 1.0 eq) was dissolved in 20 mL of dichloromethane, and then triethylamine (1.31 mL, 9.42 mmol, 2.0 eq) was added dropwise. While stirring in an ice bath, p-nitrobenzenesulfonyl chloride (1.56 g, 7.07 mmol, 1.5 eq) was slowly added. The ice bath was then removed, and the mixture was brought to room temperature. The reaction was monitored by TLC. After the reaction was complete, 20 mL of saturated sodium chloride solution was added, followed by extraction with dichloromethane (20 mL × 3 times). The organic phase was separated, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The intermediate was separated by silica gel column chromatography to obtain 72.18 g of intermediate as a white solid, with a yield of 76%. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ8.45(d,J=8.7Hz,2H,Ph-H),8.38(d,J=7.9Hz,1H,NH),8.10(d,J=8.7Hz,2H,Ph-H),7.23–7.16(m,3H,Ph-H),7.10(d,J=7.9Hz,2 H,Ph-H),6.97(d,J=8.7Hz,2H,Ph-H),6.88–6.79(m,2H,Ph-H),4.44(td,J=8.6,5.3Hz,1H,CH),3.90(d,J=16.4Hz,1H,piperazine-CH),3.83(d,J=16.0H z,1H,piperazine-CH),3.79(s,3H,OCH3),3.68(d,J=16.5Hz,1H,piperazineCH),3.63(d,J=16.4Hz,1H,piperazineCH),3.34–3.26(m,2H,piperazine -CH2),3.18(t,J=5.0Hz,2H,piperazine-CH2),3.10(s,3H,NCH3),2.84(dd,J=13.5,4.8Hz,1H,PhCH),2.62(dd,J=13.4,9.4Hz,1H,PhCH).ESI-HRMS:m / z 610.1961[M+H] + C 29 H 31 N5O8S[609.1893].

[0066] Example 9: Preparation of Intermediate 8

[0067] Intermediate 7 (2.00 g, 3.28 mmol) was dissolved in methanol:dichloromethane (15 mL:15 mL), and then 10% palladium on carbon (15.0 mg) was added. The mixture was purged three times with hydrogen gas, and stirred overnight at room temperature under hydrogen balloon protection. After the reaction was complete, diatomaceous earth was added for filtration, the solvent was evaporated under reduced pressure, and the intermediate was separated by silica gel column chromatography to obtain 81.56 g of intermediate as a white solid, with a yield of 82%. Spectroscopic data: 1HNMR (400MHz, DMSO-d6) δ8.37(d,J=7.9Hz,1H,NH),7.42(d,J=8.6Hz,2H,Ph-H),7.30–7.15(m,3H,Ph-H),7.12(d,J=7.9Hz,2H,Ph-H),6. 98(d,J=8.8Hz,2H,Ph-H),6.88–6.79(m,2H,Ph-H),6.68(d,J=8.7Hz,2H,Ph-H),6.20(s,2H,NH2),4.44(td,J=8.8,5.2Hz,1H,CH),3.86(s ,2H,piperazine-CH2),3.79(s,3H,OCH3),3.46–3.36(m,2H,piperazineCH2),3.23–3.12(m,2H,piperazine-CH2),3.10(s,3H,NCH3),3 .07–2.95(m,2H,piperazine-CH2),2.84(dd,J=13.5,4.8Hz,1H,PhCH),2.63(dd,J=13.4,9.5Hz,1H,PhCH).ESI-HRMS:m / z580.2228[M+H] + C 29 H 33 N5O6S[579.2152].

[0068] Example 10: Preparation of intermediate 9(ae)

[0069] N-tert-butoxycarbonylcarboxylic acid fragments of different lengths (0.259 mmol, 1.2 eq.) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.388 mmol, 1.5 eq.) were dissolved in dichloromethane (15 mL), and the mixture was in an ice bath for 30 min. Then, 8 (150 mg, 0.259 mmol, 1.0 eq.) and N,N-diisopropylethylamine (0.518 mmol, 2.0 eq.) were added, and the mixture was allowed to cool to room temperature after removing the ice bath. After the reaction was complete, the solvent was removed under reduced pressure, and the mixture was extracted with water (20 mL) and dichloromethane (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography to obtain intermediate 9 (ae).

[0070] The N-tert-butoxycarbonylcarboxylic acid fragment used was N-tert-butoxycarbonyl-4-aminobutyric acid. Product 9a was a yellow solid with a yield of 76%. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ10.37(s,1H,NH),8.36(d,J=8.0Hz,1H,NH),7.92–7.83(m,2H,Ph-H),7.74(d,J=8.9Hz,2H,Ph-H),7.23–7.06(m,5H,Ph- H),7.02–6.93(m,2H,Ph-H),6.83(t,J=5.3Hz,2H,Ph-H),6.81(d,J=1.8Hz,1H,NH),4.43(td,J=8.7,5.0Hz,1H,CH),3.85(s,2H,CH2),3.78(s,3 H,OCH3),3.53–3.42(m,2H,CH2),3.14(d,J=10.4Hz,4H,CH2×2),3.09(s,3H,NCH3),2.96(q,J=6.6Hz,2H,CH2),2.83(dd,J=13.5,5.0Hz,1H,Ph- CH),2.62(dd,J=13.6,9.4Hz,1H,Ph-CH),2.35(t,J=7.4Hz,2H,CH2),1.70(p,J=7.3Hz,2H,CH2),1.36(s,9H,N(CH3)3).ESI-MS:m / z765.15[M+H] + C 38 H 48 N6O9S[764.32].

[0071] The N-tert-butoxycarbonylcarboxylic acid fragment used was N-tert-butoxycarbonyl-6-aminohexanoic acid. Product 9b was a yellow solid with a yield of 78%. Spectroscopic data: ESI-MS: m / z 793.59 [M+H] + 815.69 [M+Na] + C 40 H 52 N6O9S[792.35].

[0072] The N-tert-butoxycarbonylcarboxylic acid fragment used was N-tert-butoxycarbonyl-7-aminoheptanoic acid. Product 9c was a yellow solid with a yield of 77%. Spectroscopic data: ESI-MS: m / z 829.60 [M+Na] + C 41 H 54 N6O9S[806.37].

[0073] The N-tert-butoxycarbonylcarboxylic acid fragment used was N-tert-butoxycarbonyl-8-aminooctanoic acid. The product was a yellow solid on day 9, with a yield of 80%. Spectroscopic data: ESI-MS: m / z 843.24 [M+Na] + C42 H 56 N6O9S[820.38].

[0074] The N-tert-butoxycarbonylcarboxylic acid fragment used was N-tert-butoxycarbonyl-10-aminodecanoic acid. Product 9e was a yellow solid with a yield of 82%. Spectroscopic data: ESI-MS: m / z 871.24 [M+Na] + C 44 H 60 N6O9S[848.41].

[0075] Example 11: Preparation of target compound 10(ae)

[0076] Intermediate 9(ae) (200 mg) was added to dichloromethane (16 mL), and trifluoroacetic acid (4 mL) was slowly added dropwise under stirring in an ice bath. After the addition was complete, the ice bath was removed and the mixture was brought to room temperature. The reaction was monitored by TLC. After the reaction was completed, the solvent was removed by vacuum distillation, and then saturated sodium bicarbonate solution was added to adjust the pH of the reaction solution to 7. Dichloromethane (15 mL × 3) was then added for extraction. The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under vacuum to obtain the target compound 10(ae).

[0077] 10a is a yellow solid, yield: 72%, melting point: 140-142℃. Spectroscopic data: 1 H NMR (400MHz, DMSO-d6) δ10.45(s,1H,NH),8.35(d,J=7.9Hz,1H,NH),7.87(d,J=8.7Hz,2H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.13(d d,J=24.3,7.1Hz,5H,Ph-H),6.96(d,J=8.8Hz,2H,Ph-H),6.86–6.79(m,2H,Ph-H),4.44(td,J=8.7,5.5Hz,1H,CH),3.85(s,2H,CH2), 3.78(s,3H,OCH3),3.54–3.43(m,2H,CH2),3.42–3.33(m,2H,CH2),3.14(d,J=8.8Hz,4H,CH2×2),3.09(s,3H,NCH3),2.83(dd,J=13.5 ,4.8Hz,1H,Ph-CH),2.62(q,J=8.2,6.8Hz,3H,Ph-CH,CH2),2.41(t,J=7.3Hz,2H,CH2),1.71(dt,J=14.2,6.8Hz,2H,NH2).ESI-MS:m / z 665.68[M+H] + C 33 H 40N6O7S[664.27].

[0078] 10b is a yellow solid with a yield of 69% and a melting point of 148-150℃. Spectroscopic data: 1 H NMR (400MHz, DMSO-d6) δ10.42(s,1H,NH),8.35(d,J=7.6Hz,1H,NH),7.88(d,J=8.7Hz,2H,Ph-H),7.74(d,J=8.9Hz,2H,Ph-H),7.14(dd,J=24. 5,7.7Hz,5H,Ph-H),6.97(d,J=8.9Hz,2H,Ph-H),6.82(d,J=7.6Hz,2H,Ph-H),4.47–4.40(m,1H,CH),3.85(s,2H,CH2),3.79(s,3H,OCH3),3.52 (m,4H,CH2×2),3.18–3.13(m,2H,CH2),3.09(s,3H,NCH3),2.96–2.87(m,2H,CH2),2.83(dd,J=13.2,5.3Hz,1H,Ph-CH),2.67(d,J=10.2Hz,1H, Ph-CH),2.41–2.32(m,2H,CH2),1.67–1.52(m,2H,NH2),1.39(dd,J=14.5,7.2Hz,2H,CH2),1.35–1.20(m,4H,CH2×2).ESI-MS:m / z693.61[M+H] + C 35 H 44 N6O7S[692.30].

[0079] 10c is a yellow solid, yield: 74%, melting point: 160-162℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ10.37(s,1H,NH),8.35(d,J=7.9Hz,1H,NH),7.81(dd,J=5 1.5,8.8Hz,2H,Ph-H),7.42(d,J=8.7Hz,1H,Ph-H),7.25–7.07(m,5H,Ph-H),6.97( d,J=8.8Hz,2H,Ph-H),6.87–6.80(m,2H,Ph-H),6.68(d,J=8.7Hz,1H,Ph-H),4.45( td,J=8.5,5.3Hz,1H,CH),3.86(s,2H,CH2),3.79(s,3H,OCH3),3.55–3.43(m,2H,C H2),3.40(d,J=5.7Hz,2H,CH2),3.37–3.25(m,2H,CH2),3.25–3.12(m,4H,CH2×2) ,3.10(s,3H,NCH3),3.09–2.99(m,2H,CH2),2.84(dd,J=13.5,4.9Hz,1H,Ph-CH),2 .67–2.59(m,1H,Ph-CH),2.37(t,J=7.3Hz,1H,CH),1.67–1.53(m,1H,CH),1.45–1. 34(m,2H,NH2),1.34–1.27(m,2H,CH2),1.24(m,2H,CH2).ESI-MS:m / z707.45[M+H] + C 36 H 46 N6O7S[706.31].

[0080] At 10 days, it is a yellow solid, yield: 71%, melting point: 140-142℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ10.46(s,1H,NH),8.36(d,J=8.0Hz,1H,NH),7.89(d,J=8.7Hz,2H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.14(dd,J=23.7, 7.3Hz,5H,Ph-H),6.97(d,J=8.9Hz,2H,Ph-H),6.87–6.78(m,2H,Ph-H),4.44(td,J=8.7,5.1Hz,1H,CH),3.91–3.81(m,2H,CH2),3.79(s,3H,OCH 3),3.49(d,J=5.6Hz,2H,CH2),3.15(d,J=9.2Hz,4H,CH2×2),3.10(s,3H,NCH3),2.84(dd,J=13.2,4.8Hz,1H,Ph-CH),2.80–2.71(m,2H,CH2),2. 63(dd,J=13.5,9.4Hz,1H,Ph-CH),2.38(t,J=7.3Hz,2H,CH2),1.66–1.57(m,2H,NH2),1.57–1.42(m,4H,CH2×2),1.31(s,6H,CH2×3).ESI-MS:m / z 721.51[M+H] + C 37 H 48 N6O7S[720.33].

[0081] 10e is a yellow solid, yield: 68%, melting point: 130-131℃. Spectral data: 1H NMR (400MHz, DMSO-d6) δ10.49(s,1H,NH),8.36(d,J=7.9Hz,1H,NH),7.89(d,J=8.8Hz,2H,Ph-H),7.74(d,J=8.8Hz,2H,Ph-H),7.14(dd,J=23.5,7.2 Hz,5H,Ph-H),6.97(d,J=8.9Hz,2H,Ph-H),6.88–6.79(m,2H,Ph-H),4.44(td,J=8.7,5.0Hz,1H,CH),3.84(d,J=17.7Hz,2H,CH2),3.79(s,3H,OCH3), 3.49(d,J=6.0Hz,2H,CH2),3.15(d,J=9.0Hz,4H,CH2×2),3.10(s,3H,NCH3),2.84(dd,J=13.4,5.0Hz,1H,Ph-CH),2.78–2.71(m,2H,CH2),2.63(dd,J =13.5,9.5Hz,1H,Ph-CH),2.37(t,J=7.3Hz,2H,CH2),1.66–1.56(m,2H,NH2),1.50(dt,J=22.5,6.1Hz,4H,CH2×2),1.28(s,10H,CH2×5).ESI-MS:m / z 749.55[M+H] + C 39 H 52 N6O7S[748.36].

[0082] Example 12: Preparation of target compound 11 (aj)

[0083] 10(ae) (100 mg, 1.0 eq.) and I-2 (1.1 eq.) or I-3 (1.1 eq.) were dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (2.0 eq.) was added. The reaction was carried out at 90 °C for 10 h. After the reaction was completed, the solvent was removed by vacuum distillation, and the mixture was extracted with water (15 mL) and ethyl acetate (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain the target compound 11(aj).

[0084] 11a is a yellow solid, yield: 42%, melting point: 224-225℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H,NH),10.43(s,1H,NH),8.34(d,J=7.9Hz,1H, NH),7.86(d,J=8.8Hz,2H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.61–7.54(m,1H,P h-H),7.22–7.06(m,6H,Ph-H),7.02(d,J=7.0Hz,1H,Ph-H),6.96(d,J=8.9Hz,2H, Ph-H),6.87–6.78(m,2H,Ph-H),6.67(t,J=5.8Hz,1H,NH),5.05(dd,J=12.7,5.4Hz ,1H,CH),4.43(q,J=8.6Hz,1H,CH),3.85(s,2H,CH2),3.78(s,3H,OCH3),3.54–3. 42(m,2H,CH2),3.38(q,J=6.7Hz,2H,CH2),3.14(d,J=9.3Hz,4H,CH2×2),3.09(s,3 H,NCH3),2.90–2.78(m,2H,CH2),2.66–2.54(m,2H,CH2),2.47(d,J=7.2Hz,2H,CH 2),2.12–1.93(m,2H,CH2),1.91(p,J=6.9Hz,2H,CH2).ESI-MS:m / z919.38[MH]-;C 46 H 48 N8O 11 S[920.32].

[0085] 11b is a yellow solid with a yield of 54% and a melting point of 175-177℃. Spectroscopic data: 1H NMR(400MHz,DMSO-d6)δ11.10(s,1H,NH),10.37(s,1H,NH),8.37(d,J=8.0Hz,1H,NH),7 .87(d,J=8.8Hz,2H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.61–7.53(m,1H,Ph-H),7.29–7 .06(m,6H,Ph-H),7.02(d,J=7.1Hz,1H,Ph-H),6.97(d,J=8.8Hz,2H,Ph-H),6.87–6.79( m,2H,Ph-H),6.56(t,J=5.9Hz,1H,NH),5.05(dd,J=12.8,5.2Hz,1H,CH),4.43(td,J=8.7 ,5.4Hz,1H,CH),3.85(s,2H,CH2),3.78(s,3H,OCH3),3.54–3.45(m,2H,CH2),3.29(d,J =6.4Hz,2H,CH2),3.14(d,J=9.7Hz,4H,CH2×2),3.09(s,3H,NCH3),2.92–2.79(m,2H,CH2 ),2.66–2.55(m,2H,CH2),2.38(t,J=7.2Hz,2H,CH2),2.19–1.94(m,2H,CH2),1.63(tt, J=14.6,7.6Hz,4H,CH2×2),1.39(dt,J=16.9,8.8Hz,2H,CH2).ESI-MS:m / z971.32[M+Na] + C 48 H 52 N8O 11 S[948.35].

[0086] 11c is a yellow solid, yield: 38%, melting point: 210-212℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H,NH),10.36(s,1H,NH),8.35(d,J=7.9Hz,1H,N H),7.87(d,J=8.8Hz,2H,Ph-H),7.74(d,J=8.8Hz,2H,Ph-H),7.62–7.54(m,1H,Ph- H),7.21–7.06(m,6H,Ph-H),7.02(d,J=7.0Hz,1H,Ph-H),6.97(d,J=8.8Hz,2H,Ph- H),6.87–6.79(m,2H,Ph-H),6.54(t,J=5.9Hz,1H,NH),5.05(dd,J=12.8,5.4Hz,1H, CH),4.44(td,J=8.6,5.3Hz,1H,CH),3.85(s,2H,CH2),3.78(s,3H,OCH3),3.55–3. 46(m,2H,CH2),3.31–3.26(m,2H,CH2),3.19–3.11(m,4H,CH2×2),3.10(s,3H,NCH3 ),2.91–2.80(m,2H,CH2),2.67–2.55(m,2H,CH2),2.37(t,J=7.3Hz,2H,CH2),2.09 –1.95(m,2H,CH2),1.69–1.53(m,4H,CH2×2),1.42–1.33(m,4H,CH2×2).ESI-MS:m / z 963.54 [M+H] + C 49 H 54 N8O 11 S[962.36].

[0087] 11d is a yellow solid, yield: 40%, melting point: 215-216℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H,NH),10.36(s,1H,NH),8.35(d,J=7.9Hz,1H,NH ),7.87(d,J=8.8Hz,2H,Ph-H),7.74(d,J=8.8Hz,2H,Ph-H),7.63–7.54(m,1H,Ph-H), 7.14(dd,J=23.3,6.3Hz,6H,Ph-H),7.02(d,J=7.0Hz,1H,Ph-H),6.97(d,J=8.1Hz,2 H,Ph-H),6.89–6.78(m,2H,Ph-H),6.53(t,J=5.9Hz,1H,NH),5.05(dd,J=12.8,5.4Hz ,1H,CH),4.44(td,J=8.5,5.2Hz,1H,CH),3.85(s,2H,CH2),3.78(s,3H,OCH3),3.54 –3.46(m,2H,CH2),3.28(q,J=6.4,5.7Hz,2H,CH2),3.19–3.11(m,4H,CH2×2),3.10(s ,3H,NCH3),2.86(ddd,J=17.9,9.7,4.4Hz,2H,CH2),2.67–2.56(m,2H,CH2),2.36(t ,J=7.2Hz,2H,CH2),1.69–1.50(m,4H,CH2×2),1.44–1.21(m,8H,CH2×4).ESI-MS:m / z 977.54[M+H] + C 50 H 56 N8O 11 S[976.38].

[0088] 11e is a yellow solid, yield: 35%, melting point: 210-212℃. Spectral data: 1H NMR(400MHz,DMSO-d6)δ11.09(s,1H,NH),10.34(s,1H,NH),8.35(d,J=7.9Hz,1H,NH),7 .87(d,J=8.8Hz,2H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.63–7.54(m,1H,Ph-H),7.22– 7.05(m,6H,Ph-H),7.02(d,J=7.0Hz,1H,Ph-H),6.97(d,J=7.7Hz,2H,Ph-H),6.83(d,J= 7.4Hz,2H,Ph-H),6.52(t,J=5.8Hz,1H,NH),5.05(dd,J=12.7,5.3Hz,1H,CH),4.50–4.40 (m,1H,CH),3.85(s,2H,CH2),3.79(s,3H,OCH3),3.51(q,J=6.6Hz,2H,CH2),3.45–3.37 (m,2H,CH2),3.31–3.24(m,2H,CH2),3.22–3.11(m,4H,CH2×2),3.10(s,3H,NCH3),2.84( dd,J=13.2,4.6Hz,2H,CH2),2.68–2.58(m,2H,CH2),2.40–2.31(m,2H,CH2),1.68–1.49 (m,4H,CH2×2),1.33–1.27(m,6H,CH2×3),1.24(s,4H,CH2×2).ESI-MS:m / z1005.65[M+H] + C 52 H 60 N8O 11 S[1004.41].

[0089] 11f is a yellow solid, yield: 41%, melting point: 160-165℃. Spectral data: 1H NMR (400MHz, DMSO-d6) δ10.47(s,1H,NH),8.36(d,J=7.9Hz,1H,NH),7.86(d,J= 8.8Hz,2H,Ph-H),7.74(d,J=8.8Hz,2H,Ph-H),7.64–7.55(m,1H,Ph-H),7.26–7 .06(m,6H,Ph-H),7.02(d,J=7.0Hz,1H,Ph-H),6.96(d,J=8.9Hz,2H,Ph-H),6.8 8–6.78(m,2H,Ph-H),6.69(t,J=6.3Hz,1H,NH),5.12(dd,J=13.0,5.3Hz,1H,CH) ,4.47–4.39(m,1H,CH),3.85(s,2H,CH2),3.78(s,3H,OCH3),3.48(d,J=6.4Hz, 2H,CH2),3.31(s,2H,CH2),3.16(t,J=5.2Hz,4H,CH2×2),3.14–3.10(m,2H,CH2 ),3.09(s,3H,NCH3),3.02(s,3H,NCH3),2.87–2.75(m,2H,CH2),2.66–2.59(m, 2H,CH2),2.09–1.95(m,2H,CH2),1.90(dt,J=14.6,7.3Hz,2H,CH2).ESI-MS:m / z 957.45 [M+Na] + C 47 H 50 N8O 11 S[934.33].

[0090] 11g is a yellow solid; yield: 39%; melting point: 170-172℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ10.37(s,1H,NH),8.37(d,J=7.8Hz,1H,NH),7.86(d,J=8.8Hz,2 H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.63–7.53(m,1H,Ph-H),7.14(dd,J=23.4,7.2Hz ,6H,Ph-H),7.02(d,J=7.0Hz,1H,Ph-H),6.97(d,J=8.7Hz,2H,Ph-H),6.89–6.78(m,2H, Ph-H),6.56(t,J=5.9Hz,1H,NH),5.12(dd,J=12.8,5.2Hz,1H,CH),4.49–4.39(m,1H,CH) ,3.85(s,2H,CH2),3.78(s,3H,OCH3),3.48(d,J=6.7Hz,2H,CH2),3.44–3.38(m,2H,CH2 ),3.29(d,J=6.7Hz,2H,CH2),3.22–3.11(m,4H,CH2×2),3.09(s,3H,NCH3),3.02(s,3H, NCH3),2.83(dd,J=13.0,4.5Hz,2H,CH2),2.62(dd,J=13.1,9.2Hz,2H,CH2),2.42–2.31 (m,2H,CH2),1.76–1.46(m,4H,CH2×2),1.46–1.30(m,2H,CH2).ESI-MS:m / z963.05[M+H] + C 49 H 54 N8O 11 S[962.36].

[0091] After 11 hours, it is a yellow solid; yield: 28%; melting point: 180-181℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ10.35(s,1H,NH),8.35(d,J=8.0Hz,1H,NH),7.87(d,J=8.8Hz ,2H,Ph-H),7.74(d,J=8.8Hz,2H,Ph-H),7.64–7.54(m,1H,Ph-H),7.23–7.07(m,6H,P h-H),7.02(d,J=7.0Hz,1H,Ph-H),6.97(d,J=8.7Hz,2H,Ph-H),6.87–6.79(m,2H,Ph- H),6.54(t,J=5.9Hz,1H,NH),5.12(dd,J=13.0,5.4Hz,1H,CH),4.51–4.39(m,1H,CH) ,3.85(s,2H,CH2),3.78(s,3H,OCH3),3.54–3.46(m,2H,CH2),3.41(dd,J=12.8,6.8H z,2H,CH2),3.31–3.25(m,2H,CH2),3.15(d,J=8.7Hz,4H,CH2×2),3.10(s,3H,NCH3), 3.02(s,3H,NCH3),2.87–2.76(m,2H,CH2),2.62(dt,J=16.2,8.1Hz,2H,CH2),2.37(t ,J=7.3Hz,2H,CH2),1.69–1.54(m,4H,CH2×2),1.41–1.33(m,4H,CH2×2).ESI-MS:m / z 977.29[M+H] + C 50 H 56 N8O 11 S[976.38].

[0092] 11i is a yellow solid, yield: 36%, melting point: 170-172℃. Spectral data: 1H NMR (400MHz, DMSO-d6) δ10.34(s,1H,NH),8.35(d,J=7.8Hz,1H,NH),7.87(d,J=8.8Hz ,2H,Ph-H),7.74(d,J=8.8Hz,2H,Ph-H),7.62–7.55(m,1H,Ph-H),7.22–7.07(m,6H,Ph -H),7.02(d,J=7.0Hz,1H,Ph-H),6.97(d,J=8.8Hz,2H,Ph-H),6.86–6.79(m,2H,Ph-H ),6.53(t,J=5.9Hz,1H,NH),5.12(dd,J=13.0,5.4Hz,1H,CH),4.51–4.39(m,1H,CH),3 .85(s,2H,CH2),3.78(s,3H,OCH3),3.49(d,J=6.3Hz,2H,CH2),3.41(d,J=3.6Hz,2H, CH2),3.31–3.26(m,2H,CH2),3.18–3.11(m,4H,CH2×2),3.09(s,3H,NCH3),3.02(s,3H ,NCH3),2.87–2.76(m,2H,CH2),2.62(dd,J=13.9,9.7Hz,2H,CH2),2.36(t,J=7.4Hz,2 H,CH2),1.64–1.54(m,4H,CH2×2),1.38–1.32(m,6H,CH2×3).ESI-MS:m / z991.43[M+H] + C 51 H 58 N8O 11 S[990.39].

[0093] 11j is a yellow solid, yield: 32%, melting point: 158-160℃. Spectroscopic data: 1H NMR (400MHz, DMSO-d6) δ10.34(s,1H,NH),8.35(d,J=7.8Hz,1H,NH),7.87(d,J=8.7Hz,2 H,Ph-H),7.74(d,J=8.7Hz,2H,Ph-H),7.63–7.55(m,1H,Ph-H),7.20–7.06(m,6H,Ph-H), 7.02(d,J=6.9Hz,1H,Ph-H),6.97(d,J=8.2Hz,2H,Ph-H),6.83(d,J=7.2Hz,2H,Ph-H),6. 53(t,J=5.9Hz,1H,NH),5.12(dd,J=12.9,5.4Hz,1H,CH),4.48–4.40(m,1H,CH),3.85(s, 2H,CH2),3.79(s,3H,OCH3),3.54–3.46(m,2H,CH2),3.45–3.35(m,2H,CH2),3.30–3.24( m,2H,CH2),3.20–3.11(m,4H,CH2×2),3.10(s,3H,NCH3),3.02(s,3H,NCH3),2.88–2.77( m,2H,CH2),2.62(dd,J=13.6,9.6Hz,2H,CH2),2.35(t,J=7.5Hz,2H,CH2),1.58(td,J=14 .6,7.6Hz,4H,CH2×2),1.35(d,J=3.8Hz,2H,CH2),1.33–1.26(m,8H,CH2×4).ESI-MS:m / z 1019.56[M+H] + C 53 H 62 N8O 11 S[1018.43].

[0094] Example 13. In vitro anti-HIV activity assay of the target compound (MT-4 cells)

[0095] Terminology Explanation: MT-4 cells: human acute lymphoblastic leukemia cells; MTT assay: MTT stands for 3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide, trade name: thiazolium blue; DMSO: dimethyl sulfoxide.

[0096] Test Principles

[0097] Because HIV-infected MT-4 cells undergo cytopathic effects within a certain period (5-7 days), an appropriate concentration of the test compound solution was added to an HIV-infected MT-4 cell suspension. After a period of culture (5-7 days), MT-4 cell viability was determined using the MTT assay to obtain the drug concentration (EC50) that protects 50% of cells from cytopathic effects. 50 This allows us to determine the anti-HIV activity of the target compound. Simultaneously, we can determine the concentration (CC) at which the target compound induces cytopathic effects in 50% of uninfected HIV-positive cells. 50 ), calculate the selectivity index (SI = CC) 50 / EC 50 ).

[0098] The principle of MTT assay: MTT, or 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide, binds to succinate dehydrogenase in living cells but does not react with dead cells. Currently, the MTT assay is a rapid enzyme assay method for reflecting cell viability.

[0099] Test materials and methods

[0100] (1) HIV-1(III) B HIV-2 (ROD) strain: provided by the Institute of Microbiology and Immunology, Rega Institute, University of Leuven, Belgium;

[0101] (2) MT-4 cells: provided by the Institute of Microbiology and Immunology, Rega Institute, University of Leuven, Belgium;

[0102] (3) MTT: Purchased from Sigma, Inc., USA;

[0103] (4) Sample preparation: Dissolve the sample in DMSO to prepare an appropriate concentration before use, and dilute it 5 times with double-distilled water, with 5 dilutions in total;

[0104] (5) Positive controls: I-3L, PF74;

[0105] (6) Test method: After dilution, the sample was added to the suspension of HIV-infected MT-4 cells. After a period of time, cell viability was determined by the MTT assay. The absorbance (A) value was recorded at 590 nm using an ELISA reader, and the EC50 was calculated. 50 CC 50 And SI;

[0106] (7) MTT staining method: After adding the sample and culturing for a period of time, add 20 μL of MTT solution (5 mg / mL) to each well and continue culturing for several hours. Discard the staining solution and add 150 μL of LDMSO to each well. Mix thoroughly and record the absorbance at 590 nm in an ELISA reader.

[0107] The specific steps are as follows: Dissolve the compound in DMSO or water, then dilute with phosphate buffer, adding 3×10... 5 MT-4 cells were pre-incubated with 100 μL of different concentrations of compound solutions at 37°C for 1 h. Then, 100 μL of an appropriate concentration of virus diluent was added to this mixture, and the cells were incubated at 37°C for 1 h. After washing three times, the cells were resuspended in culture media containing or without the compound. The cells were then incubated for another 7 days at 37°C in a 5% CO2 environment, and the original culture medium was replenished with either compound-containing or compound-free media on the third day post-infection. Each culture condition was repeated twice. The cytopathic effect of the virus was monitored daily using a reverse optical microscope. Generally, the virus diluents used in this experiment often resulted in cytopathic effects on the fifth day post-infection. The drug inhibitory concentration was defined as the concentration at which the drug produced 50% inhibition of the virus-induced cytopathic effect without direct cytotoxicity to the cells (EC50). 50 It is worth emphasizing that when a compound has poor water solubility and requires DMSO to dissolve, the volumetric concentration of DMSO relative to water is generally less than 10% (the final concentration of DMSO in MT-4 cell culture medium is less than 2%). Because DMSO can affect the antiviral activity of the tested compound, antiviral activity control experiments with the same concentration of DMSO solution should be performed in parallel. Furthermore, the final concentration of DMSO (1 / 1000) is far lower than the concentration required to affect HIV replication in MT-4 cells.

[0108] In vitro anti-HIV-1(III) activity of the target compound B The activity screening data for HIV-2 (ROD) were provided by the Institute of Microbiology and Immunology, Rega Institute, University of Leuven, Belgium. All activity data were obtained through at least three independent, parallel experiments, and the results are shown in Table 1.

[0109] Table 1. Anti-HIV-1 / 2 activity, toxicity, and selectivity index of the target compound (MT-4 cells)

[0110]

[0111]

[0112] a EC 50The concentration of compounds that protect 50% of HIV-1-infected cells from cytopathic effects; b CC 50 The concentration of a compound that causes 50% of uninfected HIV-1 cells to become diseased; c SI: Selectivity coefficient, CC 50 / EC 50 The ratio; I-3L, PF74: reported HIV-1 capsid protein regulators, used as positive controls.

[0113] Experimental conclusions and analysis: As shown in Table 1, the newly synthesized benzylsulfonamide piperazine glycine derivatives of this invention exhibit significant anti-HIV activity. All compounds (except 10b) showed anti-HIV-1 activity at low micromolar concentration levels, EC 10. 50 The values ​​ranged from 0.35 to 4.39 μM. Among them, 11h (EC) 50 =0.35±0.02μM) and I-3L (EC 50 =0.24±0.12μM) has comparable activity to PF74 (EC) 50 =2.47±1.36μM) 7 times; in addition, 10d (EC 50 =0.79±0.15μM), 11b (EC 50 =0.97±0.17μM), 11c(EC) 50 =0.71±0.19μM), 11d(EC) 50 (0.88±0.33 μM) also showed good activity, with most compounds in this series exhibiting better activity than PF74. Furthermore, this series of compounds possesses superior anti-HIV-2 activity, EC50... 50 The values ​​ranged from 0.02 to 1.54 μM, among which 10a (EC) 50(IIIB) =3.29±1.34μM, EC 50(ROD) =0.04±0.02μM, Ratio (IIIB / ROD) =82.25) is a selective regulator of HIV-2 capsid protein. Therefore, the newly synthesized benzylsulfonamide piperazine glycine derivative of this invention is worthy of further research.

[0114] Example 14. Effect of the target compound on HIV-1 capsid protein content

[0115] HEK293T cells (5×10) 4 (4 μg / well) of Env-deficient HIV-1pNL4-3-LucR protein carrying HIV-1CA protein was seeded in 96-well plates and incubated at 37°C for 24 hours. Five hours or 24 hours before adding the test compound, 4 μg of the compound was added. + E -The plasmid was transfected into HEK293T cells. Transfection was performed using polyethyleneimine (PEI) co-precipitation. After 5 hours of transfection and culture, the DNA-containing medium was removed, cells were washed with DMEM, and fresh medium containing 10 μL of the test compound was added. Note that for wells treated with the protease inhibitor MG-132, HEK293T cells needed to be pre-incubated with MG-132 for 1 hour before treatment with the test compound. Control wells included those without test compound treatment or without vector transfection. At different time points, each well was repeated three times to collect attached cells and supernatant, and lysed using 0.1% Triton-X (Sigma-Aldrich, St. Louis, Missouri, USA) for subsequent p24 quantification analysis by ELISA. Lysed cells can be stored at -80°C if not used directly.

[0116] Each well of an ELISA plate was coated with 2 μg / mL mouse anti-p24 and incubated overnight at 4°C. The incubation was then stopped at 25°C with 3% BSA for 2 hours, followed by washing four times with PBST buffer (0.05% Tween in PBS). Lysed cells were cultured at 25°C for 2 hours. Simultaneously, p24 protein standards were added to prepare a standard curve. After 2 hours of incubation, the ELISA plate was washed four times with PBST buffer, and 0.1 μg / mL horseradish peroxidase (HRP)-conjugated mouse anti-human HIV-1 p24 monoclonal antibody was added, followed by incubation at 25°C for 1 hour. The plate was then thoroughly washed (four times) with PBST buffer. Subsequently, 0.4 mg / mL o-phenylenediamine in citrate phosphate buffer and sodium perborate (Sigma-Aldrich) solution was added, and the plate was incubated in the dark for 30 minutes. Multiskan was then used. TM The GO Microplate spectrophotometer (purchased from Thermo Scientific) measures optical density at a wavelength of 450 nm.

[0117] The results of the effect of representative compounds on HIV-1 capsid protein content are as follows: Figure 1 As shown. Figure 1 A represents the result obtained without the addition of the proteasome inhibitor MG-132. Figure 1 B represents the result obtained after adding MG-132. Test results show that compound 11h can effectively reduce the content of HIV-1 capsid protein and preliminarily demonstrates that it may be degraded via the proteasome pathway. Therefore, this invention provides a new direction for the development of anti-HIV drugs.

Claims

1. A benzenesulfonamide piperazine glycine derivative, characterized in that, It is one of the compounds having the following structures: 。 2. The method for preparing the benzenesulfonamide piperazine ketone glycine derivative as described in claim 1, characterized in that, The synthesis steps are as follows: Intermediate I-2 was obtained by refluxing 3-fluorophthalic anhydride I-1 with 3-amino-2,6-piperidinidone hydrochloride and sodium acetate in acetic acid solvent at 120 °C for 10 h. I-2 was dissolved in N,N-dimethylformamide solution, and iodomethane was added with potassium carbonate as a base under stirring. The reaction was carried out at room temperature for 24 h to obtain intermediate I-3. Intermediate I-2 was obtained by amide condensation of N-Boc-L-phenylalanine 1 and N-methyl-4-aminoanisole in dichloromethane. Intermediate I-3 was obtained by removing the Boc group from 2 in dichloromethane under the action of trifluoroacetic acid. Intermediate I-3 was then amide condensed with bromoacetic acid in dichloromethane to obtain intermediate 4. Intermediate I-4 was then amide condensed with 1-Boc-3-piperazinone in N,N-dimethylformamide solvent under the action of cesium carbonate. N 2. A nucleophilic substitution reaction yields intermediate 5; subsequently, 5 is de-Boc-treated with trifluoroacetic acid in dichloromethane to obtain intermediate 6; then, 6 is reacted with p-nitrobenzenesulfonyl chloride in dichloromethane in triethylamine to obtain intermediate 7; 7 is then hydrogenated and reduced in methanol and dichloromethane under 10% palladium on carbon catalysis to obtain the key intermediate 8; N-tert-butoxycarbonyl carboxylic acid fragments of different lengths and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate are reacted in an ice-water bath for 30 min, and intermediate 8 and N,N-diisopropylethylamine are added and stirred overnight at room temperature to obtain intermediate 9(ae); 9(ae) is de-Boc-treated with trifluoroacetic acid in dichloromethane to obtain the target compound 10(ae); 10(ae) is reacted with I-2 or I-3 in N, In N-dimethylformamide solution, using N,N-diisopropylethylamine as a base, reflux at 90 °C for 10 h to obtain target compound 11 (ae) or 11 (fj). The synthesis route is as follows: Reagents and conditions: (i) 3-Amino-2,6-piperidinidone hydrochloride, acetic acid, sodium acetate, 120 °C, 10 h; (ii) Potassium carbonate, iodomethane, N,N-dimethylformamide, room temperature, 24 h; (iii) N-methyl-4-aminoanisole, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, 0 °C → room temperature; (iv) Trifluoroacetic acid, dichloromethane, 0 °C → room temperature; (v) Bromoacetic acid, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, 0 °C → room temperature; (vi) 1-Boc-3-piperazinone, cesium carbonate, N,N-dimethylformamide, 45℃; (vii) trifluoroacetic acid, dichloromethane, 0℃→room temperature; (viii) p-nitrobenzenesulfonyl chloride, triethylamine, dichloromethane, 0℃→room temperature; (ix) H2, 10% Pd•C, dichloromethane / methanol, room temperature; (x) N-tert-butoxycarbonylcarboxylic acid fragments of different lengths, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, 0℃, 30 min, room temperature; (xi) trifluoroacetic acid, dichloromethane, 0℃→room temperature; (xii) I-2 or I-3, N,N-dimethylformamide, N,N-diisopropylethylamine, 90℃, 10 h; The N-tert-butoxycarbonyl carboxylic acid fragments of different lengths include: N-tert-butoxycarbonyl-4-aminobutyric acid, N-tert-butoxycarbonyl-6-aminohexanoic acid, N-tert-butoxycarbonyl-7-aminoheptanoic acid, N-tert-butoxycarbonyl-8-aminooctanoic acid, and N-tert-butoxycarbonyl-10-aminodecanoic acid.

3. The preparation method of the benzenesulfonamide piperazine ketone glycine derivative as described in claim 2, wherein the specific steps are as follows: (1) Dissolve 3-fluorophthalic anhydride I-1, 3-amino-2,6-piperidindione hydrochloride and sodium acetate in acetic acid and reflux at 120 °C for 10 h. After the reaction is completed, remove the solvent under reduced pressure, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and obtain I-2 after separation and purification by silica gel column chromatography. (2) I-2 and potassium carbonate were added to N,N-dimethylformamide, and then iodomethane was added dropwise. The mixture was stirred at room temperature for 24 hours. After the reaction was completed, the solvent was removed by vacuum distillation, the mixture was extracted with ethyl acetate, the organic phase was dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated under vacuum, and the crude product was purified by silica gel column chromatography to obtain I-3. (3) Boc-L-phenylalanine 1,2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate was added to dichloromethane and stirred for 30 min under ice bath conditions; then N,N-diisopropylethylamine and N-methyl-4-aminoanisole were added to the reaction solution, the ice bath was removed and the mixture was brought to room temperature, and TLC was used for monitoring; after the reaction was completed, the solvent was removed by vacuum distillation and the mixture was extracted with dichloromethane; the organic phase was dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated under vacuum, and the crude product was purified by silica gel column chromatography to obtain intermediate 2; (4) Add intermediate 2 obtained in the previous step to dichloromethane. Under ice bath conditions and stirring, slowly add trifluoroacetic acid to this solution. After the addition is complete, remove the ice bath and bring to room temperature. Monitor the reaction by TLC. After the reaction is complete, remove the solvent under reduced pressure. Then add saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7. Then add dichloromethane solution for extraction. Dry the organic phase with anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain intermediate 3. (5) Bromoacetic acid and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate were added to dichloromethane and stirred in an ice bath for 30 min. Then, intermediate 3 and N,N-diisopropylethylamine were added to this solution, the ice bath was removed and the mixture was brought to room temperature, and TLC was used for monitoring. After the reaction was completed, the solvent was removed by vacuum distillation and the mixture was extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain intermediate 4. (6) Intermediate 4, 1-Boc-3-piperazinone and cesium carbonate were added to N,N-dimethylformamide and stirred at 45°C for 12 h after the reaction was completed. After the reaction was completed, an appropriate amount of saturated sodium chloride solution was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain intermediate 5. (7) Add intermediate 5 to dichloromethane and slowly add trifluoroacetic acid dropwise under stirring in an ice bath. After the addition is complete, remove the ice bath and bring the mixture to room temperature. Monitor the reaction by TLC. After the reaction is complete, remove the solvent under reduced pressure and then add saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7. Then add dichloromethane solution for extraction. Dry the organic phase with anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain intermediate 6. (8) Add intermediate 6 and triethylamine to dichloromethane, and slowly add p-nitrobenzenesulfonyl chloride under stirring in an ice bath. Then remove the ice bath and bring to room temperature. Monitor by TLC. After the reaction is complete, add saturated sodium chloride solution, extract with dichloromethane, dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and obtain intermediate 7 after separation and purification by silica gel column chromatography. (9) Dissolve intermediate 7 in methanol and dichloromethane, add 10% palladium on carbon, replace with hydrogen three times, stir overnight at room temperature under hydrogen balloon protection; after the reaction is complete, add diatomaceous earth for filtration, evaporate the filtrate to dryness to obtain intermediate 8; (10) N-tert-butoxycarbonylcarboxylic acid fragments of different lengths and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate were added to dichloromethane and stirred in an ice bath for 30 min. Then, intermediate 8 and N,N-diisopropylethylamine were added to the reaction solution, the ice bath was removed and the mixture was brought to room temperature, and TLC was used for monitoring. After the reaction was completed, the solvent was removed by vacuum distillation and the mixture was extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain intermediate 9 (ae). (11) Intermediate 9 (ae) was dissolved in dichloromethane, and trifluoroacetic acid was slowly added dropwise under stirring in an ice bath. After the addition was complete, the ice bath was removed and the mixture was brought to room temperature. The reaction was monitored by TLC. After the reaction was completed, the solvent was removed by vacuum distillation, and then saturated sodium bicarbonate solution was added to adjust the pH of the reaction solution to 7. Then, dichloromethane solution was added for extraction. The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by silica gel column chromatography to obtain the target compound 10 (ae). (12) Dissolve 10 (ae) and I-2 or I-3 in N,N-dimethylformamide, add N,N-diisopropylethylamine, and react at 90 °C for 10 h. After the reaction is complete, remove the solvent under reduced pressure, add water and ethyl acetate for extraction, dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and obtain the target compound 11 (aj) by silica gel column chromatography.

4. The use of the benzylsulfonamide piperazine glycine derivative as described in claim 1 in the preparation of drugs for the treatment and prevention of AIDS.

5. An anti-HIV drug composition, characterized in that, It comprises the benzylsulfonamide piperazine glycine derivative of claim 1 or one or more pharmaceutically acceptable carriers or excipients.

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