CN109824756B - Phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone and preparation method and application thereof - Google Patents

Abstract

The invention provides a phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone and a preparation method and application thereof. The derivative has a structure shown in the following general formula I. The invention also relates to a preparation method of the derivatives and application of the derivatives as HIV-1 inhibitors in preparing anti-AIDS drugs.

Description

Phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone and preparation method and application thereof

Technical Field

The invention belongs to the technical field of organic compound synthesis and medical application, and particularly relates to a phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone, and a preparation method and application thereof.

Background

Acquired Immune Deficiency Syndrome (AIDS) is a serious infectious disease that endangers Human life and health, mainly caused by Human Immunodeficiency Virus Type I (HIV-1). Currently, clinically applied drugs for treating aids mainly include: reverse transcriptase inhibitors, protease inhibitors, integrase inhibitors and invasion inhibitors. The high-efficiency Antiretroviral Therapy (HAART) prolongs the survival time of patients to a great extent and improves the life quality of the patients, but the problems of drug resistance, drug toxic and side effects, latent infection, high cost of long-term taking of drugs and the like greatly reduce the efficacy of the Therapy, limit the application of the Therapy and further force researchers to develop anti-AIDS drugs with new targets, new mechanisms and new structures.

The HIV-1 capsid is assembled from a portion of the Gag precursor protein that is cleaved to yield capsid protein units. During the conversion of immature virions to mature virions, capsid proteins assemble into capsids, encapsulating viral RNA and nuclear-associated proteins (reverse transcriptase, protease, integrase, etc.) to form mature HIV-1 virions. The mature virus particle is infectious and the next round of replication of the virus can take place. In recent years, with the researchers' deep knowledge of capsid protein structure, the related information of crystal structure is reported in succession. Therefore, the capsid protein of HIV-1 can be used as the action target of a novel anti-HIV-1 medicament.

The Pfizer company obtains a compound PF-74 capable of obviously inhibiting HIV-1 replication through high-throughput screening of a compound library, and the structure-activity relationship and mechanism research of the compound shows that the compound interferes with the uncoating process of viruses and the process of forming infectious particles by combining HIV-1 capsid protein. Although PF-74 has novel structure, unique mechanism and definite target, PF-74 has lower curative effect, poorer drug-like property and is easy to induce drug resistance compared with the anti-HIV-1 drug on the market at present. Therefore, the development of capsid protein inhibitors with higher efficacy and good drug-like and drug-resistance is an attractive direction in the development field of anti-aids drugs in recent years.

According to the crystal structure characteristics of the binding site of the PF-74 and HIV-1 capsid protein, the phenylalanine HIV-1 capsid protein inhibitor of 4- (phenylsulfonyl) piperazine-2-ketone with a brand-new structure is discovered through reasonable drug design, chemical synthesis and biological activity evaluation, and the problems of low curative effect, poor drug property and drug resistance of the existing HIV-1 capsid protein inhibitor are hopefully improved.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone and a preparation method thereof, and also provides an activity screening result of the compound as an HIV-1 capsid protein inhibitor and an application thereof.

The technical scheme of the invention is as follows:

1. phenylalanine derivatives containing 4- (phenylsulfonyl) piperazin-2-one

A phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one, or a pharmaceutically acceptable salt, ester, or prodrug thereof, having the structure shown in formula I:

wherein the content of the first and second substances,

r is: c₁-C₆Alkyl, OC₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₃-C₆Cycloalkyl, OC₃-C₆Cycloalkyl, substituted benzene ring, substituted benzyl, substituted naphthalene ring, various substituted six-membered heterocyclic rings or various substituted five-membered heterocyclic rings, H, OH, halogen, nitro, amino, cyano, trifluoromethyl or trifluoromethoxy.

In accordance with a preferred aspect of the present invention,

r is: H. 4-F, 3-F, 2-F, 4-Cl, 3-Cl, 2-Cl, 4-Br, 3-Br, 2-Br, 4-methoxy, 3-methoxy, 2-methoxy, 4-methyl, 3-methyl, 2-methyl, 4-nitro, 3-nitro, 2-nitro, 4-amino, 3-amino and 2-amino.

According to a further preferred embodiment of the invention, the phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one is one of the following compounds:

as used herein, "pharmaceutically acceptable salts" means salts of the compounds which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and lower animals without undue toxicity, irritation, and allergic response and the like, are commensurate with a reasonable benefit-to-risk ratio, are generally water or oil soluble or dispersible, and are effective for their intended use. Including pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts, which are contemplated herein and are compatible with the chemical nature of the compounds of formula I. A list of suitable salts is found on pages 1-19 of s.m. berge et al, j.pharm.sci.,1977, 66.

The term "prodrug" as used herein refers to pharmaceutically acceptable derivatives such that the resulting biotransformation product of these derivatives is the active drug as defined for the compound of formula I.

2. Preparation method of phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone

A method for preparing a phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one comprising the steps of: generating an intermediate 2 with Boc-L-phenylalanine 1 as an initial raw material and dichloromethane as a reaction solvent through an amide condensation reaction and N-methyl-4-aminoanisole; then the intermediate 2 is dissolved in a proper amount of dichloromethane, and Boc groups are removed under the action of trifluoroacetic acid to obtain an intermediate 3; then, carrying out amide condensation reaction on the intermediate 3 and bromoacetic acid to obtain an intermediate 4; the intermediate 4 and 3-oxo-1-piperazine carboxylic acid tert-butyl ester take nucleophilic substitution reaction by taking dichloromethane as a solvent to obtain an intermediate 5; then, dissolving the intermediate 5 in a proper amount of dichloromethane solution, and removing Boc groups under the action of trifluoroacetic acid to obtain an intermediate 6; finally, the intermediate 6 and the corresponding substituted benzene sulfonyl chloride are subjected to sulfonylation reaction to obtain a target compound (7 a-k); the compound (7i-k) is further reduced by hydrogenation to give the objective compound (7 l-n).

The synthetic route is as follows:

reagents and conditions: (i) n-methyl-4-aminoanisole, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, N, N-diisopropylethylamine and dichloromethane are added, and the temperature is changed to room temperature at 0 ℃; (ii) trifluoroacetic acid, dichloromethane, room temperature; (iii) bromoacetic acid, O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate, N, N-diisopropylethylamine and dichloromethane, and the temperature is changed to room temperature at 0 ℃; (iv) 3-oxo-1-piperazine carboxylic acid tert-butyl ester, potassium carbonate and N, N-dimethylformamide, and the temperature is changed to 40 ℃ at room temperature; (v) trifluoroacetic acid, dichloromethane, room temperature; (vi) correspondingly substituted benzene sulfonyl chloride, triethylamine and dichloromethane are cooled to room temperature at 0 ℃; (vii) h₂10% Pd. C, dichloromethane/methanol, room temperature.

Wherein R is as described in formula I above.

The substituted benzene sulfonyl chloride is benzene sulfonyl chloride, 4-fluorobenzene sulfonyl chloride, 3-fluorobenzene sulfonyl chloride, 2-fluorobenzene sulfonyl chloride, 4-chlorobenzene sulfonyl chloride, 4-bromobenzene sulfonyl chloride, 4-methoxybenzene sulfonyl chloride, 4-methylbenzene sulfonyl chloride, 4-nitrobenzene sulfonyl chloride, 3-nitrobenzene sulfonyl chloride and 2-nitrobenzene sulfonyl chloride.

The room temperature of the invention is 20-30 ℃.

The preferred preparation method of the phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone comprises the following steps:

(1) adding Boc-L-phenylalanine 1 and 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate into dichloromethane, and stirring for 30min under an ice bath condition; adding N, N-diisopropylethylamine and N-methyl-4-aminoanisole into the reaction solution, removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, evaporating the solvent under reduced pressure, then adding a saturated sodium bicarbonate solution into the residue in the bottle, extracting with dichloromethane, separating an organic phase, adding a 1N HCl solution for washing, separating the organic phase, adding a saturated sodium chloride solution for washing, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and separating and purifying the obtained crude product by silica gel column chromatography to obtain an intermediate 2;

(2) adding the intermediate 2 obtained in the previous step into dichloromethane, slowly dropwise adding excessive trifluoroacetic acid into the solution under the condition of ice bath and stirring, then removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, evaporating the solvent under reduced pressure, adding a saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7, and then adding a dichloromethane solution for extraction; separating and taking an organic phase, washing the organic phase for 3 times by using a saturated sodium chloride solution, drying the organic phase by using anhydrous sodium sulfate, filtering, and evaporating the solvent to dryness under reduced pressure to obtain an intermediate 3;

(3) adding bromoacetic acid and O- (7-azabenzotriazole-1-yl) -N, N, N ', N' -tetramethylurea hexafluorophosphate into dichloromethane, and stirring for 1h under an ice bath condition; then adding the intermediate 3 and N, N-diisopropylethylamine into the solution, removing the ice bath, and stirring at room temperature for 6 h; after the reaction is finished, filtering, decompressing and evaporating the solvent, and separating by silica gel column chromatography to obtain an intermediate 4;

(4) adding the intermediate 4, the tert-butyl 3-oxo-1-piperazinecarboxylate and potassium carbonate into N, N-dimethylformamide, and stirring for 12 hours from room temperature to 40 ℃; after the reaction is finished, adding a proper amount of saturated sodium chloride solution into the reaction solution, extracting with ethyl acetate, drying an organic phase with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure, and separating by silica gel column chromatography to obtain an intermediate 5;

(5) adding the intermediate 5 into dichloromethane, slowly adding trifluoroacetic acid into the dichloromethane under the condition of ice bath and stirring, then removing the ice bath, transferring the mixture to room temperature, and monitoring by TLC; after the reaction is finished, evaporating the solvent under reduced pressure, adding a saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7, and then adding a dichloromethane solution for extraction; separating and taking an organic phase, washing the organic phase for 3 times by using a saturated sodium chloride solution, drying the organic phase by using anhydrous sodium sulfate, filtering, and evaporating the solvent to dryness under reduced pressure to obtain an intermediate 6;

(6) adding the intermediate 6 and triethylamine into dichloromethane, slowly adding the corresponding substituted benzenesulfonyl chloride into the dichloromethane under the condition of ice bath and stirring, removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, adding saturated sodium chloride solution, extracting by dichloromethane, separating and taking an organic phase, drying by using anhydrous sodium sulfate, filtering, evaporating the solvent by decompression, separating by silica gel column chromatography to obtain a crude product of the target compound, and recrystallizing by ethyl acetate to obtain a pure product (7a-k) of the target compound;

(7) dissolving a target compound (7i-k) in methanol and dichloromethane, adding 10% Pd & C, replacing with hydrogen for three times, and stirring at room temperature for two hours under the protection of a hydrogen balloon; after the reaction is finished, adding diatomite for filtration, evaporating the filtrate to dryness to obtain a crude product of the target compound, and purifying by a silica gel preparation plate to obtain a pure product (7l-7n) of the target compound.

3. Use of phenylalanine derivatives containing 4- (phenylsulfonyl) piperazin-2-one

The invention discloses a screening result of anti-HIV-1 activity of a phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone and an application thereof as an HIV-1 inhibitor for the first time. Experiments prove that the phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone can be used as an HIV-1 inhibitor for preparing anti-AIDS medicaments. The invention also provides application of the compound in preparing anti-HIV-1 medicines.

anti-HIV-1 Activity and toxicity test of the target Compound

A class of phenylalanine derivatives containing 4- (phenylsulfonyl) piperazin-2-one synthesized as described above were tested for anti-HIV-1 activity and toxicity at the cellular level, and their anti-HIV-1 activity and toxicity data are shown in Table 1, with the literature-reported capsid protein inhibitor PF-74 as a positive control.

The newly synthesized phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone shows remarkable anti-HIV-1 activity. For example, the anti-HIV-1 activity of the target compounds 7a, 7c, 7d, 7f, 7h, 7i, 7l, 7M, 7n was in the range of 0.09-0.54. mu.M, which was superior to that of the positive control PF-74 (EC)₅₀＝0.52±0.18μM,CC₅₀>47.00,SI>90.91) or equivalent, wherein the target compoundsanti-HIV-1 Activity of substance 7l (EC)₅₀＝0.09±0.03μM,CC₅₀>34.50,SI>383.36) has great significance in further research.

The phenylalanine derivatives containing 4- (benzenesulfonyl) piperazine-2-ketone can be used as HIV-1 inhibitors. In particular to the application of the compound as an HIV-1 inhibitor in preparing anti-AIDS drugs.

An anti-HIV-1 pharmaceutical composition comprises a class of phenylalanine derivatives containing 4- (phenylsulfonyl) piperazin-2-one of the present invention and one or more pharmaceutically acceptable carriers or excipients.

The invention provides a phenylalanine derivative containing 4- (benzenesulfonyl) piperazine-2-ketone and a preparation method thereof, and also provides a screening result of anti-HIV-1 activity of partial compounds and the first application thereof in the field of antivirus. Tests prove that the phenylalanine derivatives containing 4- (benzenesulfonyl) piperazine-2-one can be used as HIV-1 inhibitors and have high application value. In particular to the application of the compound as an HIV-1 inhibitor in preparing anti-AIDS drugs.

Detailed Description

The invention will be understood by the following examples, which are given by way of illustration and are not intended to limit the scope of the invention.

Example 1: preparation of tert-butyl (S) - (1- ((4-methoxyphenyl) (methyl) amino) -1-oxo-3-phenylpropan-2-yl) carbamate (2)

The starting material Boc-L-phenylalanine (1) (2.90g,10.93mmol,1.5eq.) and 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate (5.69g,10.93mmol,1.5eq) were added to 20mL of dichloromethane and stirred under ice bath conditions for 30 min; then N, N-diisopropylethylamine (3.61mL,21.87mmol,3eq.) and N-methyl-4-aminoanisole (1.0g, 7.29mmol,1eq.) were added, the ice bath was removed and the mixture was stirred at room temperature and monitored by TLC; after 6h, the reaction was completed, the solvent was evaporated under reduced pressure, then 40mL of saturated sodium bicarbonate solution and 40mL of dichloromethane were added to the residue in the flask for extraction, the organic phase was separated and washed with 40mL of 1N HCl solution, the organic phase was separated and washed with 40mL of saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product of tert-butyl (2) intermediate (S) - (1- ((4-methoxyphenyl) (methyl) amino) -1-oxo-3-phenylpropan-2-yl) carbamate (2), 2.48g of which was a yellow oily substance, in 89% yield, which was obtained by silica gel column chromatography (eluent EA: PE ═ 1: 8).

Spectral data:

¹H 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),4.27–4.06(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).

¹³C NMR(100MHz,DMSO-d₆)δ172.22(C＝O),158.98,155.75(C＝O),138.53(2×C),136.12(2×C),129.28(2×C),128.47(2×C),126.70,115.21(2×C),78.33,55.94,53.55,37.86,37.07,28.65(3×C).

ESI-MS:m/z 385.4(M+1),407.5(M+23).C₂₂H₂₈N₂O₄[384.5].

example 2: preparation of (S) -2-amino-N- (4-methoxyphenyl) -N-methyl-3-phenylpropionamide (3)

Intermediate 2(4.0g,10.40mmol,1.0eq.) was added to 30mL of dichloromethane, and trifluoroacetic acid (3.86mL,52.02mmol,5.0eq.) was then added slowly to this solution, stirred at room temperature, monitored by TLC; after 1h, the reaction was completed, and then the reaction solution was adjusted to pH 7 with saturated sodium bicarbonate solution, extracted with 40mL of dichloromethane, the organic phase was separated, washed with saturated sodium chloride solution (20 mL. times.3 times), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 2.36g of a crude product of intermediate (S) -2-amino-N- (4-methoxyphenyl) -N-methyl-3-phenylacrylamide (3), as a yellow oil, in 80% yield.

Spectral 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).

¹³C NMR(100MHz,DMSO-d₆)δ174.89(C＝O),158.75,139.00,136.35,129.51(2×C),128.93(2×C),128.47(2×C),126.55,115.04(2×C),55.85,53.35,42.19,37.45.

ESI-MS:m/z 285.05(M+1).C₁₇H₂₀N₂O₂[284.36].

example 3: preparation of intermediate (S) -2- (2-bromoacetyl) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropanamide (4)

Bromoacetic acid (117mg,0.84mmol,1.2eq.), O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate (401mg,1.06mmol,1.5eq.) were added to 15mL dichloromethane and stirred under ice bath conditions for 1 h; to this solution was then added intermediate 3(200mg,0.70mmol,1eq.) and N, N-diisopropylethylamine (232 μ L,1.41mmol,2eq.), removed from the ice bath, stirred at room temperature, monitored by TLC; after completion of the reaction for 6 hours, the solvent was distilled off under reduced pressure, and the residue was separated by column chromatography on silica gel (eluent EA: PE ═ 1:4+ 2.5% triethylamine) to give 190mg of intermediate (S) -2- (2-bromoacetyl) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropionamide (4) as a white oil in 68% yield.

Spectral data:

ESI-MS:m/z 405.4(M+1).C₁₉H₂₁BrN₂O₃[404.1].

example 4: preparation of intermediate (S) -tert-butyl 4- (2- ((1- ((4-methoxyphenyl) (methyl) amino) -1-oxo-3-phenylpropan-2-yl) amino) -2-oxoethyl) -3-tert-butyloxopiperazine-1-carboxylate (5)

Intermediate 4(700mg,1.73mmol,1eq.) and tert-butyl 3-oxo-1-piperazinecarboxylate (415mg,2.07mmol,1.2eq.), potassium carbonate (477mg,3.45mmol,2eq.) were added to N, N-dimethylformamide (6mL) and stirred from room temperature to 40 ℃ for 12 h; after the reaction, 20mL of a saturated sodium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20mL × 3 times), the organic phase was dried over anhydrous sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the intermediate (S) -tert-butyl 4- (2- ((1- ((4-methoxyphenyl) (methyl) amino) -1-oxo-3-phenylpropan-2-yl) amino) -2-oxoethyl) -3-tert-butyloxopiperazine-1-carboxylate (5) was obtained by silica gel column chromatography in 360mg of a white solid, yield 40%, melting point: 138 ℃ and 139 ℃.

Spectral data:

ESI-MS:m/z 523.09(M-1).C₂₈H₃₆N₄O₆[524.62].

example 5: preparation of intermediate (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (2-oxopiperazin-1-yl) acetylamino) -3-phenylpropanamide (6)

Intermediate 5(158mg,0.301mmol) was added to dichloromethane (5mL) and trifluoroacetic acid (2mL) was added slowly thereto with stirring under ice bath conditions, followed by removal of the ice bath to room temperature and monitoring by TLC; after 4h, after the reaction is finished, the solvent is evaporated under reduced pressure, then a saturated sodium bicarbonate solution is added to adjust the pH of the reaction solution to 7, and then 20mL of dichloromethane solution is added for extraction; the organic phase was separated, washed with saturated sodium chloride solution (20mL × 3 times), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain 80mg of intermediate (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (2-oxopiperazin-1-yl) acetylamino) -3-phenylpropionamide (6) as a yellow oil with a yield of 63%.

Spectral data:

ESI-MS:m/z 425.4(M+1),447.4(M+23).C₂₃H₂₈N₄O₄[424.5].

example 6: preparation of target Compound (7a-k)

Dissolving the intermediate 6(1eq.) and triethylamine (2eq.) in 10mL of dichloromethane, slowly adding the corresponding substituted benzenesulfonyl chloride (1.5eq.) while stirring in an ice bath, removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, 20mL of saturated sodium chloride solution is added, then dichloromethane is added for extraction (20mL multiplied by 3 times), an organic phase is separated, dried by anhydrous sodium sulfate, filtered, and the solvent is evaporated to dryness under reduced pressure to obtain a crude product of the target compound, and the crude product of the target compound is purified by a silica gel preparation plate or recrystallized by ethyl acetate to obtain a pure product (7a-k) of the target compound.

Reaction of the correspondingly substituted benzenesulfonyl chloride with benzenesulfonyl chloride (45 μ L,0.35mmol,1.5eq.) with intermediate 6(100mg,0.24mmol, 1eq.), triethylamine (65 μ L,0.47mmol,2eq.) gave 65mg of (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (2-oxo-4- (phenylsulfonyl) piperazin-1-yl) acetylamino) -3-phenylpropanamide (7a) as a white powdery solid in 49% yield, melting point: 84-85 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.38(d,J＝7.9Hz,1H,NH),7.87–7.73(m,3H,Ph-H),7.68(t,J＝7.5Hz,2H,Ph-H),7.22–7.14(m,3H,Ph-H),7.11(d,J＝8.1Hz,2H,Ph-H),6.97(d,J＝8.8Hz,2H,Ph-H),6.87–6.77(m,2H,Ph-H),4.43(td,J＝8.8,5.1Hz,1H,CH),3.88–3.80(m,2H,piperazine-CH₂),3.78(s,3H,OCH₃),3.55(d,J＝16.2Hz,1H,piperazineCH),3.49(d,J＝16.2Hz,1H,piperazineCH),3.25–3.11(m,4H,piperazine-CH₂×2),3.09(s,3H,NCH₃),2.83(dd,J＝13.5,4.8Hz,1H,PhCH),2.62(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.36(C＝O),167.32(C＝O),163.62(C＝O),159.00,137.90,135.99,134.83,134.18,130.12(2×C),129.28(2×C),129.12(2×C),128.59(2×C),128.12(2×C),126.88,115.13(2×C),55.89,51.82,48.99,48.41,46.95,43.20,37.77,37.68.

ESI-HRMS:m/z 565.2112(M+1),1151.3958(2M+23).C₂₉H₃₂N₄O₆S[564.2043].

reaction of the correspondingly substituted benzenesulfonyl chloride with 4-fluorobenzenesulfonyl chloride (103mg,0.53mmol,1.5eq.) with intermediate 6(150mg,0.35mmol, 1eq.), triethylamine (98 μ L,0.71mmol,2eq.) gave 73mg of (S) -2- (2- (4- ((4-fluorophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropanamide (7b) as a white solid in 35% yield, m.p.: 199 ℃ and 200 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.38(d,J＝7.9Hz,1H,NH),7.90(dd,J＝8.6,5.2Hz,2H,Ph-H),7.52(t,J＝8.7Hz,2H,Ph-H),7.24–7.14(m,3H,Ph-H),7.11(d,J＝8.0Hz,2H,Ph-H),6.97(d,J＝8.7Hz,2H,Ph-H),6.90–6.75(m,2H,Ph-H),4.43(td,J＝8.7,5.1Hz,1H,CH),3.88(d,J＝16.5Hz,1H,piperazine-CH),3.85(d,J＝16.4Hz,1H,piperazine-CH),3.79(s,3H,OCH₃),3.58(d,J＝16.3Hz,1H,piperazine-CH),3.52(d,J＝16.2Hz,1H,piperazine-CH),3.28–3.18(m,2H,piperazine-CH₂),3.16(m,2H,piperazine-CH₂),3.10(s,3H,NCH₃),2.84(dd,J＝13.4,4.7Hz,1H,PhCH),2.62(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.32(C＝O),165.41(d,¹J_CF＝250.9Hz),163.62(C＝O),159.00,137.90,135.89,131.35(2×C),131.25,129.28(2×C),129.12(2×C),128.59(2×C),126.88,117.45,117.22,115.12(2×C),55.88,51.82,48.93,48.40,46.91,43.13,37.76,37.69.

ESI-HRMS:m/z 583.2020(M+1),1187.3740(2M+23).C₂₉H₃₁FN₄O₆S[582.1948].

reaction of the correspondingly substituted benzenesulfonyl chloride with 3-fluorobenzenesulfonyl chloride (75 μ L,0.57mmol,1.5eq.) with intermediate 6(160mg,0.38mmol, 1eq.), triethylamine (104 μ L,0.75mmol,2eq.) gave 90mg of (S) -2- (2- (4- ((3-fluorophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropanamide (7c) as a white solid in 41% yield, m.p.: 182 ℃ and 183 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.38(d,J＝7.9Hz,1H,NH),7.81–7.57(m,4H,Ph-H),7.24–7.14(m,3H,Ph-H),7.11(d,J＝7.9Hz,2H,Ph-H),6.97(d,J＝8.7Hz,2H,Ph-H),6.82(d,J＝6.6Hz,2H,Ph-H),4.43(td,J＝8.7,5.3Hz,1H,CH),3.88(d,J＝16.6Hz,1H,piperazine-CH),3.83(d,J＝16.8Hz,1H,piperazine-CH),3.79(s,3H,OCH₃),3.62(d,J＝16.2Hz,1H,piperazine-CH),3.56(d,J＝16.3Hz,1H,piperazine-CH),3.29–3.21(m,2H,piperazine-CH₂),3.20–3.12(m,2H,piperazine-CH₂),3.10(s,3H,NCH₃),2.84(dd,J＝13.5,4.7Hz,1H,PhCH),2.62(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.33(C＝O),163.60(C＝O),162.47(d,¹J_CF＝247.6Hz),159.00,137.90,137.12(d,³J_CF＝6.8Hz),135.90,132.48(d,³J_CF＝7.9Hz),129.28(2×C),129.13(2×C),128.59(2×C),126.88,124.41(d,⁴J_CF＝3.0Hz),121.35(d,²J_CF＝21.0Hz),115.34,115.13(2×C),55.88,51.83,48.89,48.42,46.95,43.17,37.76,37.68.

ESI-HRMS:m/z 583.2017(M+1),1187.3844(M+23).C₂₉H₃₁FN₄O₆S[582.1948].

reaction of the corresponding substituted benzenesulfonyl chloride with 2-fluorobenzenesulfonyl chloride (103mg,0.53mmol,1.5eq.) with intermediate 6(150mg,0.35mmol, 1eq.), triethylamine (98 μ L,0.71mmol,2eq.) gave 91mg of (S) -2- (2- (4- ((2-fluorophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropanamide (7d) as a white solid in 44% yield, m.p.: 170 ℃ and 171 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.39(d,J＝7.9Hz,1H,NH),7.91–7.75(m,2H,Ph-H),7.60–7.50(m,1H,Ph-H),7.47(t,J＝7.6Hz,1H,Ph-H),7.23–7.15(m,3H,Ph-H),7.12(d,J＝7.8Hz,2H,Ph-H),6.98(d,J＝8.7Hz,2H,Ph-H),6.88–6.80(m,2H,Ph-H),4.45(td,J＝8.7,5.2Hz,1H,CH),3.91(d,J＝16.5Hz,1H,piperazine-CH),3.86(d,J＝16.6Hz,1H,piperazine-CH),3.80(s,3H,OCH₃),3.73(d,J＝16.3Hz,1H,piperazine-CH),3.70(d,J＝16.2Hz,1H,piperazine-CH),3.38(t,J＝4.9Hz,2H,piperazine-CH₂),3.24–3.13(m,2H,piperazine-CH₂),3.11(s,3H,NCH₃),2.85(dd,J＝13.5,4.8Hz,1H,PhCH),2.63(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.34(C＝O),163.69(C＝O),159.00,158.82(d,¹J_CF＝252.5Hz),137.91,136.92(d,³J_CF＝8.7Hz),135.91,131.44,129.28(2×C),129.13(2×C),128.59(2×C),126.88,125.81(d,⁴J_CF＝3.5Hz),124.15(d,²J_CF＝14.5Hz),118.24(d,²J_CF＝21.5Hz),115.14(2×C),55.89,51.83,48.47,48.43,47.11,42.79,37.77,37.70.

ESI-HRMS:m/z 583.2017(M+1),1187.3790(2M+23).C₂₉H₃₁FN₄O₆S[582.1948].

reaction of the correspondingly substituted benzenesulfonyl chloride with 4-chlorobenzenesulfonyl chloride (112mg,0.53mmol,1.5eq.) with intermediate 6(150mg,0.35mmol, 1eq.), triethylamine (98 μ L,0.71mmol,2eq.) gave 93mg of (S) -2- (2- (4- ((4-chlorophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropionamide (7e) as a white solid in 44% yield, melting point: 203 ℃ and 204 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.38(d,J＝8.0Hz,1H,NH),7.83(d,J＝8.6Hz,2H,Ph-H),7.74(d,J＝8.6Hz,2H,Ph-H),7.25–7.14(m,3H,Ph-H),7.11(d,J＝8.0Hz,2H,Ph-H),6.97(d,J＝8.8Hz,2H,Ph-H),6.88–6.74(m,2H,Ph-H),4.44(td,J＝8.7,5.2Hz,1H,CH),3.88(d,J＝16.5Hz,1H,piperazine-CH),3.83(d,J＝16.6Hz,1H,piperazine-CH),3.79(s,3H,OCH₃),3.59(d,J＝16.2Hz,1H,piperazineCH),3.53(d,J＝16.2Hz,1H,piperazineCH),3.28–3.18(m,2H,piperazine-CH₂),3.18–3.12(m,2H,piperazine-CH₂),3.10(s,3H,NCH₃),2.84(dd,J＝13.4,4.8Hz,1H,PhCH),2.62(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.32(C＝O),163.59(C＝O),159.00,139.16,137.90,135.89,133.91,130.25(2×C),130.05(2×C),129.28(2×C),129.12(2×C),128.59(2×C),126.88,115.13(2×C),55.88,51.81,48.87,48.43,46.94,43.09,37.77,37.69.

ESI-HRMS:m/z 599.1725(M+1),1219.3204(2M+23).C₂₉H₃₁ClN₄O₆S[598.1653].

reaction of the corresponding substituted benzenesulfonyl chloride with 4-bromobenzenesulfonyl chloride (145mg,0.57mmol,1.5eq.) with intermediate 6(160mg,0.38mmol, 1eq.), triethylamine (104 μ L,0.75mmol,2eq.) gave 75mg of (S) -2- (2- (4- ((4-bromophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetylamino) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropanamide (7f) as a white solid in 31% yield, m.p.: 204 ℃ and 205 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.42(d,J＝8.0Hz,1H,NH),7.94(d,J＝8.5Hz,2H,Ph-H),7.80(d,J＝8.5Hz,2H,Ph-H),7.30–7.20(m,3H,Ph-H),7.17(d,J＝8.0Hz,2H,Ph-H),7.03(d,J＝8.8Hz,2H,Ph-H),6.94–6.83(m,2H,Ph-H),4.50(td,J＝8.7,5.2Hz,1H,CH),3.94(d,J＝16.5Hz,1H,piperazine-CH),3.89(d,J＝16.7Hz,1H,piperazine-CH),3.84(s,3H,OCH₃),3.65(d,J＝16.2Hz,1H,piperazineCH),3.59(d,J＝16.2Hz,1H,piperazineCH),3.33–3.24(m,2H,piperazine-CH₂),3.24–3.19(m,2H,piperazine-CH₂),3.15(s,3H,NCH₃),2.90(dd,J＝13.5,4.8Hz,1H,PhCH),2.68(dd,J＝13.4,9.4Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.32(C＝O),163.59(C＝O),159.00,137.90,135.89,134.30,133.19(2×C),130.10(2×C),129.28(2×C),129.13(2×C),128.59(2×C),128.25,126.88,115.13(2×C),55.89,51.82,48.86,48.43,46.95,43.08,37.77,37.68.

ESI-HRMS:m/z 643.1224(M+1).C₂₉H₃₁BrN₄O₆S[642.1148].

reaction of the correspondingly substituted benzenesulfonyl chloride with 4-methoxybenzenesulfonyl chloride (117mg,0.57mmol,1.5eq.) with intermediate 6(160mg,0.38mmol, 1eq.), triethylamine (104 μ L,0.75mmol,2eq.) produced 89mg of (S) -2- (2- (4- ((4-methoxyphenyl) sulfonyl) -2-oxopiperazin-1-yl) acetylamino) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropanamide (7g) as a white solid in 40% yield, m.p.: 199 ℃ and 200 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.38(d,J＝8.0Hz,1H,NH),7.75(d,J＝8.8Hz,2H,Ph-H),7.25–7.14(m,5H,Ph-H),7.11(d,J＝7.8Hz,2H,Ph-H),6.98(d,J＝8.8Hz,2H,Ph-H),6.88–6.78(m,2H,Ph-H),4.44(td,J＝8.7,5.1Hz,1H,CH),3.95–3.83(m,5H,PhOCH₃+piperazine-CH₂),3.79(s,3H,OCH₃),3.51(d,J＝16.2Hz,1H,piperazine-CH),3.45(d,J＝16.2Hz,1H,piperazine-CH),3.22–3.15(m,2H,piperazine-CH₂),3.15–3.12(m,2H,piperazine-CH₂),3.10(s,3H,NCH₃),2.84(dd,J＝13.5,4.8Hz,1H,PhCH),2.63(dd,J＝13.4,9.4Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.36(C＝O),167.33(C＝O),163.71,163.62(C＝O),159.00,137.90,135.89,130.47(2×C),129.28(2×C),129.12(2×C),128.59(2×C),126.87,126.09,115.26(2×C),115.13(2×C),56.25,55.88,51.81,49.08,48.39,46.93,43.23,37.77,37.70.

ESI-HRMS:m/z 595.2226(M+1),1211.4210(2M+23).C₃₀H₃₄N₄O₇S[594.2148].

the corresponding substituted benzenesulfonyl chloride was reacted with 4-methylbenzenesulfonyl chloride (101mg,0.53mmol,1.5eq.) with intermediate 6(150mg,0.35mmol, 1eq.), triethylamine (98 μ L,0.71mmol,2eq.) to give 132mg of (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (2-oxo-4-tolylpiperazin-1-yl) acetylamino) -3-phenylpropanamide (7h) as a white solid in 65% yield, m.p.: 206- & lt207 & gt.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.35(d,J＝8.0Hz,1H,NH),7.70(d,J＝8.1Hz,2H,Ph-H),7.48(d,J＝8.1Hz,2H,Ph-H),7.21–7.14(m,3H,Ph-H),7.10(d,J＝7.9Hz,2H,Ph-H),6.97(d,J＝8.8Hz,2H,Ph-H),6.87–6.76(m,2H,Ph-H),4.44(td,J＝8.7,5.1Hz,1H,CH),3.87(d,J＝16.5Hz,1H,piperazine-CH),3.82(d,J＝17.7Hz,1H,piperazine-CH),3.78(s,3H,OCH₃),3.52(d,J＝16.2Hz,1H,piperazineCH),3.46(d,J＝16.2Hz,1H,piperazineCH),3.21–3.12(m,4H,piperazine-CH₂×2),3.09(s,3H,NCH₃),2.83(dd,J＝13.5,4.9Hz,1H,PhCH),2.62(dd,J＝13.4,9.4Hz,1H,PhCH),2.42(s,3H,PhCH₃).

¹³C NMR(100MHz,DMSO-d₆)δ171.35(C＝O),167.32(C＝O),163.65(C＝O),158.99,144.72,137.90,135.89,131.80,130.55(2×C),129.28(2×C),129.12(2×C),128.58(2×C),128.20(2×C),126.87,115.13(2×C),55.88,51.80,49.01,48.41,46.94,43.21,37.76,37.70,21.53.

ESI-HRMS:m/z 579.2275(M+1),1179.4327(2M+23).C₃₀H₃₄N₄O₆S[578.2199].

reaction of the corresponding substituted benzenesulfonyl chloride with 4-nitrobenzenesulfonyl chloride (212mg,0.95mmol,1.5eq.) with intermediate 6(270mg,0.64mmol, 1eq.), triethylamine (176 μ L,1.27mmol,2eq.) gave 264mg of (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (4- ((4-nitrophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -3-phenylpropanamide (7i) as a white solid in 68% yield, melting point: 212 ℃ and 213 ℃.

Spectral 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,2H,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＝16.0Hz,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).

¹³C NMR(100MHz,DMSO-d₆)δ171.34(C＝O),167.29(C＝O),163.44(C＝O),158.99,150.75,140.99,137.89,135.88,129.71(2×C),129.28(2×C),129.10(2×C),128.59(2×C),126.88,125.32(2×C),115.12(2×C),55.88,51.81,48.69,48.40,46.92,43.06,37.76,37.72

ESI-HRMS:m/z 610.1961(M+1),632.1739(M+23).C₂₉H₃₁N₅O₈S[609.1893].

the corresponding substituted benzenesulfonyl chloride was reacted with 3-nitrobenzenesulfonyl chloride (212mg,0.95mmol,1.5eq.) with intermediate 6(270mg,0.64mmol, 1eq.), triethylamine (176 μ L,1.27mmol,2eq.) to give 277mg of (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (4- ((3-nitrophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -3-phenylpropanamide (7j) as a white solid in 71% yield and melting point: 159 ℃ to 160 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.62–8.53(m,1H,Ph-H),8.50–8.42(m,1H,Ph-H),8.39(d,J＝7.9Hz,1H,NH),8.26(d,J＝7.9Hz,1H,Ph-H),7.97(t,J＝8.0Hz,1H,Ph-H),7.26–7.14(m,3H,Ph-H),7.11(d,J＝7.9Hz,2H,Ph-H),6.97(d,J＝8.8Hz,2H,Ph-H),6.87–6.76(m,2H,Ph-H),4.43(td,J＝8.7,5.3Hz,1H,CH),3.90(d,J＝16.4Hz,1H,piperazine-CH),3.83(d,J＝16.7Hz,1H,piperazine-CH),3.79(s,3H,OCH₃),3.70(d,J＝16.3Hz,1H,piperazineCH),3.64(d,J＝16.4Hz,1H,piperazineCH),3.34–3.26(m,2H,piperazine-CH₂),3.19(t,J＝5.1Hz,2H,piperazine-CH₂),3.10(s,3H,NCH₃),2.84(dd,J＝13.5,4.9Hz,1H,PhCH),2.62(dd,J＝13.4,9.4Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.35(C＝O),167.30(C＝O),163.48(C＝O),158.99,148.74,137.89,136.83,135.89,134.01,132.15,129.28(2×C),129.11(2×C),128.63,128.59(2×C),126.88,122.86,115.12(2×C),55.89,51.82,48.74,48.36,46.92,43.11,37.76,37.71.

ESI-HRMS:m/z 610.1970(M+1),1241.3681(2M+23).C₂₉H₃₁N₅O₈S[609.1893].

the corresponding substituted benzenesulfonyl chloride was reacted with 2-nitrobenzenesulfonyl chloride (188mg,0.85mmol,1.5eq.) with intermediate 6(240mg,0.57mmol, 1eq.), triethylamine (157 μ L,1.13mmol,2eq.) to give (S) -N- (4-methoxyphenyl) -N-methyl-2- (2- (4- ((2-nitrophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -3-phenylpropanamide (7k)205mg as a white solid in 59% yield, melting point: 160 ℃ and 161 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.40(d,J＝8.0Hz,1H,NH),8.09(d,J＝7.6Hz,1H,Ph-H),8.04(d,J＝7.7Hz,1H,Ph-H),7.95(t,J＝7.3Hz,1H,Ph-H),7.88(t,J＝7.4Hz,1H,Ph-H),7.23–7.15(m,3H,Ph-H),7.12(d,J＝8.1Hz,2H,Ph-H),6.98(d,J＝8.8Hz,2H,Ph-H),6.84(d,J＝7.1Hz,2H,Ph-H),4.46(td,J＝8.7,5.2Hz,1H,CH),3.94(d,J＝16.5Hz,1H,piperazine-CH),3.88(d,J＝16.6Hz,1H,piperazine-CH),3.85(s,2H,piperazineCH₂),3.79(s,3H,OCH₃),3.49(t,J＝5.2Hz,2H,piperazine-CH₂),3.20(t,J＝5.1Hz,2H,piperazine-CH₂),3.11(s,3H,NCH₃),2.86(dd,J＝13.5,4.8Hz,1H,PhCH),2.64(dd,J＝13.4,9.4Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.36(C＝O),167.34(C＝O),163.60(C＝O),159.00,148.34,137.91,135.90,135.65,133.08,131.07,129.29(2×C),129.21,129.13,128.60(2×C),126.89,124.88(2×C),115.14(2×C),55.89,51.83,48.54,48.47,47.26,43.00,37.77,37.73.

ESI-HRMS:m/z 610.1965(M+1),1241.3695(2M+23).C₂₉H₃₁N₅O₈S[609.1893].

example 7: preparation of target Compound (7l-n)

Dissolve 7i (150mg,0.246mmol) in methanol: dichloromethane (8mL:8mL), then 10% Pd.C (18mg) is added, hydrogen is replaced for three times, and the mixture is stirred for two hours at room temperature under the protection of a hydrogen balloon; after the reaction, adding diatomite for filtration, evaporating the filtrate to dryness to obtain a crude target compound, and purifying the crude target compound by using a silica gel preparation plate to obtain 87mg of a pure target compound (S) -2- (2- (4- ((4-aminophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetylamino) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropionamide (7l) as a white solid with the yield of 61% and the melting point: 128-129 ℃.

Spectral data:

¹H NMR(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).

¹³C NMR(100MHz,DMSO-d₆)δ171.38(C＝O),167.38(C＝O),163.98(C＝O),158.99,154.17,137.91,135.90,130.31(2×C),129.29(2×C),129.14(2×C),128.58(2×C),126.88,118.49,115.14(2×C),113.30(2×C),55.88,51.83,49.30,48.38,46.94,43.32,37.77,37.66.

ESI-HRMS:m/z 580.2228(M+1),1181.4192(2M+23).C₂₉H₃₃N₅O₆S[579.2152].

dissolving 7j (150mg,0.246mmol) in methanol-dichloromethane (5mL:5mL), adding 10% Pd.C (20mg), replacing with hydrogen for three times, and stirring at room temperature for two hours under the protection of a hydrogen balloon; after the reaction, adding diatomite for filtration, evaporating the filtrate to dryness to obtain a crude target compound, and purifying by a silica gel preparation plate to obtain 101mg of a pure target compound (S) -2- (2- (4- ((3-aminophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetylamino) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropionamide (7m), wherein the yield is 71%, the melting point is: 106-107 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.39(d,J＝8.0Hz,1H,NH),7.28(t,J＝7.9Hz,1H,Ph-H),7.24–7.15(m,3H,Ph-H),7.12(d,J＝7.7Hz,2H,Ph-H),7.02–6.93(m,3H,Ph-H),6.93–6.79(m,4H,Ph-H),5.71(s,2H,NH₂),4.44(td,J＝8.8,5.2Hz,1H,CH),3.86(s,2H,piperazine-CH₂),3.79(s,3H,OCH₃),3.50(d,J＝16.2Hz,1H,piperazineCH),3.45(d,J＝16.3Hz,1H,piperazineCH),3.23–3.11(m,4H,piperazine-CH₂×2),3.10(s,3H,NCH₃),2.84(dd,J＝13.5,4.7Hz,1H,PhCH),2.63(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.35(C＝O),163.72(C＝O),159.00,150.26,137.91,135.90,134.84,130.43,129.28(2×C),129.14(2×C),128.59(2×C),126.88,118.83,115.14(2×C),114.54,112.15,55.88,51.83,49.15,48.44,47.00,43.31,37.77,37.66.

ESI-HRMS:m/z 580.2220(M+1),1181.4181(2M+23).C₂₉H₃₃N₅O₆S[579.2152].

dissolve 7k (150mg,0.246mmol) in methanol: dichloromethane (5mL:5mL), then 10% Pd.C (20mg) is added, hydrogen is replaced for three times, and the mixture is stirred for two hours at room temperature under the protection of a hydrogen balloon; after the reaction is finished, adding diatomite for filtration, evaporating the filtrate to dryness to obtain a crude target compound, and purifying by a silica gel preparation plate to obtain a pure target compound (S) -2- (2- (4- ((2-aminophenyl) sulfonyl) -2-oxopiperazin-1-yl) acetamido) -N- (4-methoxyphenyl) -N-methyl-3-phenylpropionamide (7N)114mg, a white solid, the yield of 80%, and the melting point: 95-96 ℃.

Spectral data:

¹H NMR(400MHz,DMSO-d₆)δ8.37(d,J＝8.0Hz,1H,NH),7.44(d,J＝7.9Hz,1H,Ph-H),7.36(t,J＝7.6Hz,1H,Ph-H),7.22–7.15(m,3H,Ph-H),7.12(d,J＝7.7Hz,2H,Ph-H),6.98(d,J＝8.8Hz,2H,Ph-H),6.90(d,J＝8.3Hz,1H,Ph-H),6.87–6.79(m,2H,Ph-H),6.68(t,J＝7.5Hz,1H,Ph-H),6.14(s,2H,NH₂),4.45(td,J＝8.7,5.2Hz,1H,CH),3.86(s,2H,piperazine-CH₂),3.79(s,3H,OCH₃),3.61(d,J＝16.3Hz,1H,piperazineCH),3.56(d,J＝16.3Hz,1H,piperazineCH),3.29–3.20(m,2H,piperazine-CH₂),3.20–3.13(m,2H,piperazine-CH₂),3.10(s,3H,NCH₃),2.84(dd,J＝13.5,4.8Hz,1H,PhCH),2.64(dd,J＝13.4,9.5Hz,1H,PhCH).

¹³C NMR(100MHz,DMSO-d₆)δ171.37(C＝O),167.36(C＝O),163.85(C＝O),159.00,148.26,137.91,135.90,135.17,130.39,129.28(2×C),129.13(2×C),128.58(2×C),126.88,118.02,115.92,115.14(2×C),114.39,55.89,51.83,48.91,48.42,46.94,43.14,37.77,37.68.

ESI-HRMS:m/z 580.2225(M+1),1181.4212(2M+23).C₂₉H₃₃N₅O₆S[579.2152].

example 8 in vitro anti-HIV-1 Activity test (TZM-bl cells) of Compounds of interest

The principle is as follows: luciferase reporter Gene experiments (nef Gene deleted HIV-1NL4-3)

The test method comprises the following steps:

anti-HIV-1 infection assay in TZM-bl cells

The inhibitory activity of a compound against HIV-1 infection was determined as the degree of decrease in the expression level of luciferase gene following a single round of viral infection of TZM-bl cells. Briefly, 800TCID was used in the presence of different concentrations of compounds (7a-7n and PF-74)₅₀The virus of (NL4-3) infects TZM-bl cells. 1 day after infection, the culture medium was removed and 100. mu.L of Bright Glo reagent (Promega, San Luis Obispo, Calif.) was added to each well before use of the Victor 2 luminometerAnd detecting the fluorescence activity of the fluorescent probe. Effective concentration of compound (EC) for inhibiting HIV-1 strain₅₀) Defined as the concentration that results in a 50% decrease in luciferase activity (relative light units) compared to the virus control wells.

Cytotoxicity assays

Cytotoxicity of the synthesized compounds was determined using the CytoTox-Glo fluorescent cytotoxicity kit (purchased from Promega). TZM-bl cells were cultured for 1 day in the presence of different concentrations of compounds (7a-7n and PF-74), as determined in parallel with the anti-HIV-1 activity assay. The cytotoxicity (CC) of the tested target compound is then determined according to the procedure required by the kit₅₀) I.e., the concentration of the compound of interest required to reduce cell survival by 50%.

TABLE 1 phenylalanine derivatives containing in part 4- (phenylsulfonyl) piperazin-2-one anti-HIV-1 Activity, toxicity and selection index (TZM-bl cells)

^aEC₅₀: concentration of compound that protects 50% of HIV-1 infected cells from cytopathic effects;

^bCC₅₀: (ii) concentration of compound that causes lesions in 50% of cells not infected with HIV-1;

^cand (3) SI: coefficient of selectivity, CC₅₀/EC₅₀The ratio of (A) to (B);

PF-74: a class of HIV-1 capsid inhibitors has been reported as positive controls.

And (4) experimental conclusion analysis: as shown in Table 1, the newly synthesized phenylalanine derivatives containing 4- (phenylsulfonyl) piperazin-2-one of the present inventionThe organisms exhibit significant anti-HIV-1 activity. For example, the anti-HIV-1 activity of the target compounds 7a, 7c, 7d, 7f, 7h, 7i, 7l, 7M, 7n was in the range of 0.09-0.54. mu.M, which was superior to that of the positive control PF-74 (EC)₅₀＝0.52±0.18μM,CC₅₀>47.00,SI>90.91) or equivalent, wherein the anti-HIV-1 activity (EC) of 7l of the compound of interest₅₀＝0.09±0.03μM,CC₅₀>34.50,SI>383.36) has great significance in further research.

Claims (5)

1. A phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one, characterized in that the compound is one of the following:

2. the method for preparing a phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one according to claim 1, comprising the steps of: generating an intermediate 2 with Boc-L-phenylalanine 1 as an initial raw material and dichloromethane as a reaction solvent through an amide condensation reaction and N-methyl-4-aminoanisole; then the intermediate 2 is dissolved in a proper amount of dichloromethane, and Boc groups are removed under the action of trifluoroacetic acid to obtain an intermediate 3; then, carrying out amide condensation reaction on the intermediate 3 and bromoacetic acid to obtain an intermediate 4; the intermediate 4 and 3-oxo-1-piperazine carboxylic acid tert-butyl ester take nucleophilic substitution reaction by taking dichloromethane as a solvent to obtain an intermediate 5; then, dissolving the intermediate 5 in a proper amount of dichloromethane solution, and removing Boc groups under the action of trifluoroacetic acid to obtain an intermediate 6; finally, the intermediate 6 and the corresponding substituted benzene sulfonyl chloride are subjected to sulfonylation reaction to obtain a target compound (7a, 7c, 7d, 7f, 7h, 7i, 7j and 7 k); further hydrogenating and reducing the compound (7i-k) to obtain a target compound (7 l-n);

the synthetic route is as follows:

reagents and conditions: (i) n-methyl-4-aminoanisole, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, N, N-diisopropylethylamine and dichloromethane are added, and the temperature is changed to room temperature at 0 ℃; (ii) trifluoroacetic acid, dichloromethane, room temperature; (iii) bromoacetic acid, O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate, N, N-diisopropylethylamine and dichloromethane, and the temperature is changed to room temperature at 0 ℃; (iv) 3-oxo-1-piperazine carboxylic acid tert-butyl ester, potassium carbonate and N, N-dimethylformamide, and the temperature is changed to 40 ℃ at room temperature; (v) trifluoroacetic acid, dichloromethane, room temperature; (vi) correspondingly substituted benzene sulfonyl chloride, triethylamine and dichloromethane are cooled to room temperature at 0 ℃; (vii) h₂10% Pd · C, dichloromethane/methanol, room temperature;

wherein R is H, 3-F, 2-F, 4-Br, 4-CH₃、4-NO₂、4-NO₂、3-NO₂、2-NO₂；

The substituted benzene sulfonyl chloride is benzene sulfonyl chloride, 3-fluorobenzene sulfonyl chloride, 2-fluorobenzene sulfonyl chloride, 4-bromobenzene sulfonyl chloride, 4-methylbenzene sulfonyl chloride, 4-nitrobenzene sulfonyl chloride, 3-nitrobenzene sulfonyl chloride and 2-nitrobenzene sulfonyl chloride.

3. The method for preparing a phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one according to claim 2, comprising the steps of:

(1) adding Boc-L-phenylalanine 1 and 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate into dichloromethane, and stirring for 30min under an ice bath condition; adding N, N-diisopropylethylamine and N-methyl-4-aminoanisole into the reaction solution, removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, evaporating the solvent under reduced pressure, then adding a saturated sodium bicarbonate solution into the residue in the bottle, extracting with dichloromethane, separating an organic phase, adding a 1N HCl solution for washing, separating the organic phase, adding a saturated sodium chloride solution for washing, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and separating and purifying the obtained crude product by silica gel column chromatography to obtain an intermediate 2;

(2) adding the intermediate 2 obtained in the previous step into dichloromethane, slowly dropwise adding excessive trifluoroacetic acid into the solution under the condition of ice bath and stirring, then removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, evaporating the solvent under reduced pressure, adding a saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7, and then adding a dichloromethane solution for extraction; separating and taking an organic phase, washing the organic phase for 3 times by using a saturated sodium chloride solution, drying the organic phase by using anhydrous sodium sulfate, filtering, and evaporating the solvent to dryness under reduced pressure to obtain an intermediate 3;

(3) adding bromoacetic acid and O- (7-azabenzotriazole-1-yl) -N, N, N ', N' -tetramethylurea hexafluorophosphate into dichloromethane, and stirring for 1h under an ice bath condition; then adding the intermediate 3 and N, N-diisopropylethylamine into the solution, removing the ice bath, and stirring at room temperature for 6 h; after the reaction is finished, filtering, decompressing and evaporating the solvent, and separating by silica gel column chromatography to obtain an intermediate 4;

(4) adding the intermediate 4, the tert-butyl 3-oxo-1-piperazinecarboxylate and potassium carbonate into N, N-dimethylformamide, and stirring for 12 hours from room temperature to 40 ℃; after the reaction is finished, adding a proper amount of saturated sodium chloride solution into the reaction solution, extracting with ethyl acetate, drying an organic phase with anhydrous sodium sulfate, filtering, evaporating the solvent under reduced pressure, and separating by silica gel column chromatography to obtain an intermediate 5;

(5) adding the intermediate 5 into dichloromethane, slowly adding trifluoroacetic acid into the dichloromethane under the condition of ice bath and stirring, then removing the ice bath, transferring the mixture to room temperature, and monitoring by TLC; after the reaction is finished, evaporating the solvent under reduced pressure, adding a saturated sodium bicarbonate solution to adjust the pH of the reaction solution to 7, and then adding a dichloromethane solution for extraction; separating and taking an organic phase, washing the organic phase for 3 times by using a saturated sodium chloride solution, drying the organic phase by using anhydrous sodium sulfate, filtering, and evaporating the solvent to dryness under reduced pressure to obtain an intermediate 6;

(6) adding the intermediate 6 and triethylamine into dichloromethane, slowly adding the corresponding substituted benzenesulfonyl chloride into the dichloromethane under the condition of ice bath and stirring, removing the ice bath, transferring to room temperature, and monitoring by TLC; after the reaction is finished, adding saturated sodium chloride solution, extracting by dichloromethane, separating and taking an organic phase, drying by using anhydrous sodium sulfate, filtering, evaporating the solvent by decompression, separating by silica gel column chromatography to obtain a crude product of the target compound, and recrystallizing by using ethyl acetate to obtain pure products (7a, 7c, 7d, 7f, 7h and 7i-k) of the target compound;

(7) dissolving a target compound (7i-k) in methanol and dichloromethane, adding 10% Pd & C, replacing with hydrogen for three times, and stirring at room temperature for two hours under the protection of a hydrogen balloon; after the reaction is finished, adding diatomite for filtration, evaporating the filtrate to dryness to obtain a crude product of the target compound, and purifying by a silica gel preparation plate to obtain a pure product (7l-n) of the target compound.

4. The use of the phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one according to claim 1 for the preparation of a medicament for the treatment and prevention of aids.

5. An anti-HIV pharmaceutical composition comprising a phenylalanine derivative containing 4- (phenylsulfonyl) piperazin-2-one or a pharmaceutically acceptable salt thereof according to claim 1 and one or more pharmaceutically acceptable carriers or excipients.