Application of aminocyclobutene compound in anti-herpes simplex virus type 1 (HSV-1) medicine

Abstract

The invention relates to the technical field of medicinal chemistry, and particularly provides application of an aminocyclobutene compound in anti-herpes simplex virus type 1 (HSV-1) medicine. The compound has the structural characteristics as shown in a general formula (I), and EWG is an electron withdrawing group and is selected from aryl-substituted sulfonyl or alkyl-substituted sulfonyl; r is selected from hydrogen, alkyl, alkoxy, halogen or nitryl and can form a monosubstituted or disubstituted structure on an aromatic ring. Through systematic in-vitro activity evaluation, it is found that the aminocyclobutene compound can be used as an active ingredient for developing a novel anti-HSV-1 infection treatment drug, and the aminocyclobutene compound has a good application prospect in the aspect of preparing anti-HSV-1 virus drugs. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and particularly relates to an aminocyclobutene compound, a preparation method thereof, and an application thereof in drugs against herpes simplex virus type 1 (HSV-1). Background Art

[0002] As a privileged scaffold with significant biological functions, aminocyclobutene compounds are widely present in various bioactive natural products and drug molecules. Its unique four-membered ring strain system endows such compounds with excellent reactivity, making them show important value in the construction of complex molecules and being often used as key intermediates for the synthesis of polysubstituted cyclobutanes and functionalized amine compounds. Traditional synthetic strategies mainly rely on the Ficini [2+2] cycloaddition reaction, that is, the cycloaddition reaction of alkynamides with cyclic electron-deficient enones under high-temperature conditions to prepare aminocyclobutenes. However, such methods generally have technical bottlenecks such as harsh reaction conditions, large fluctuations in product yields, and limited substrate scope. In recent years, although the introduction of transition metal catalytic systems has improved the reaction efficiency to a certain extent, there are still two major technical barriers: First, the use of precious metal catalysts significantly increases the synthesis cost; Second, the problem of heavy metal residues leads to reduced environmental compatibility, restricting its application in drug synthesis. Therefore, developing a green and economical non-metal catalytic system has become a technical problem that urgently needs to be broken through in this field. Based on the applicant's previous research on the synthesis of aminocyclobutene compounds (Org. Chem. Front., 2023, 10, 4043–4054) and the research on the anti-tumor, anti-SARS-CoV-2, and anti-dengue virus type II activities of some aminocyclobutene compounds (see patents CN 202210171146.7; CN202210851374.9), in order to better expand the uses of aminocyclobutene and its analogs, the applicant further conducted in-depth research on the anti-herpes simplex virus type 1 (HSV-1) activity of the synthesized aminocyclobutene compounds, and found lead compounds with better anti-herpes simplex virus type 1 (HSV-1) virus activity and very low toxicity, providing lead compounds with clinical transformation potential for the development of new antiviral drugs. Summary of the Invention

[0003] Based on the aminocyclobutene compound synthesis technology developed in the early stage, through systematic structure-activity relationship research, the present invention for the first time discovers that aminocyclobutene derivatives with specific electronic effects show significant inhibitory effects on the HSV-1 virus strain. It is particularly worth pointing out that, on the premise of maintaining the low cytotoxicity of the compound, its antiviral activity index is significantly better than that of the clinical control drug acyclovir, providing a breakthrough candidate molecule for the development of new anti-herpes virus drugs.

[0004] For this reason, the object of the present invention is to provide the use of aminocyclobutene compounds in anti-herpes simplex virus type 1 (HSV-1).

[0005] To achieve the object of the present invention, the specific technical solutions are as follows:

[0006] The general structural formula of the aminocyclobutene compound is:

[0007]

[0008] Among them, EWG is an electron-withdrawing group, selected from sulfonyl group, aryl-substituted sulfonyl group or alkyl-substituted sulfonyl group; R is a substituent, selected from hydrogen, alkyl group, alkoxy group, halogen or nitro group, and R is mono-substituted or di-substituted at the ortho / meta / para position of the aromatic benzene ring.

[0009] Preferably: EWG is an electron-withdrawing group, selected from sulfonyl group, aryl-substituted sulfonyl group or C1-3 alkyl-substituted sulfonyl group, and the aryl group is phenyl substituted by methyl or methoxy group; R is selected from hydrogen, C1-3 alkyl group, C1-3 alkoxy group, halogen or nitro group, and R is mono-substituted at the ortho / meta / para position of the benzene ring.

[0010] The synthesis route of the aminocyclobutene compound is as follows:

[0011]

[0012] The specific synthesis steps are as follows:

[0013] Place the alkynylamide 1 in a dry reaction flask, replace the air with nitrogen and heat it to the molten state, and maintain this temperature until the reaction is complete as monitored by TLC. The reaction solution is directly separated by silica gel column chromatography to obtain the aminocyclobutene compound 2.

[0014] Furthermore, in the above technical solution, the synthesis route of the raw material alkynylamide 1 is as follows:

[0015]

[0016] The specific synthesis steps are as follows:

[0017] Under nitrogen protection, suspend the sulfonamide 3 and cesium carbonate in anhydrous N,N-dimethylformamide, and slowly dropwise add the anhydrous dichloromethane solution of the TMS-EBX iodide 4 under ice bath and light protection. The system is raised to room temperature and stirred until the reaction is complete, filtered through diatomaceous earth, rotary evaporated under reduced pressure and purified by column chromatography to obtain the raw material alkynylamide 1.

[0018] In the above steps, the molar ratio of the sulfonamide 3, cesium carbonate and the TMS-EBX iodide 4 is 1:1.3:1.5; the volume ratio of N,N-dimethylformamide and dichloromethane is 1:2.5.

[0019] Furthermore, in the above technical solution, through in vitro active cell experiments, it is verified that aminocyclobutene compounds not only show good anti-herpes simplex virus type 1 activity, but also have low cytotoxicity, indicating that they can be applied to the preparation of drugs for treating anti-herpes simplex virus type 1.

[0020] Compared with the prior art, the application of the aminocyclobutene compounds provided by the present invention in anti-herpes simplex virus type 1 drugs shows significant advantages: on the one hand, these compounds have good anti-herpes simplex virus type 1 activity; on the other hand, their low cytotoxicity ensures good drug safety. Based on the above characteristics, their application in the preparation of drugs for treating anti-herpes simplex virus type 1 has important development value and application prospects. Detailed implementation manners

[0021] To better understand the technical solution of the present invention, the present invention will be described in detail below through specific examples. It should be particularly noted that the following examples are only used to exemplarily illustrate the technical solution of the present invention and do not constitute a limitation on the protection scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

[0022] Main instruments and chemical reagents

[0023] Nuclear magnetic resonance spectrometer: Bruker Ascend TM Model 400; High-resolution mass spectrometer: Bruker MicrOTOF-Q II type mass; Fourier transform infrared spectrometer: Zhiwei Smart type (Tianjin Gangdong Technology Co., Ltd.); Ultraviolet analyzer: WFH-203B type (Shanghai Huyue Ming Scientific Instrument Co., Ltd.); Microscopic melting point determinator: XT4A type (Beijing Keyi Dian Guang Instrument Factory).

[0024] The reagents and materials used in the implementation process of this application can all be obtained through commercial channels.

[0025] Example 1: Synthesis of alkynylamide intermediates 1a-1o

[0026] It should be particularly noted that in the compound structure: Ts represents p-toluenesulfonyl, Mbs represents p-methoxybenzenesulfonyl, and Ms represents methylsulfonyl.

[0027]

[0028] Taking the specific synthesis procedure of alkynylamide 1a as an example, in a 25 mL dry reaction flask, sulfonamide 3a (123.7 mg, 0.50 mmol) and cesium carbonate (211.8 mg, 0.65 mmol) were successively added. After three nitrogen replacements, anhydrous N,N-dimethylformamide (1.0 mL) was added. After stirring at room temperature for 30 min, a dichloromethane solution (2.5 mL) containing TMS-EBX iodide 4 (258.2 mg, 0.75 mmol) was slowly added dropwise under light-shielded conditions at 0 °C. After the addition was complete, the temperature was slowly raised to room temperature and stirring was continued for 30 min. The reaction progress was monitored by TLC. After the raw materials were completely converted, filtration was carried out through silica gel, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography (gradient elution: petroleum ether / ethyl acetate = 10:1 → 4:1) to obtain white solid 1a (135.2 mg, 0.498 mmol), and the isolated yield was 99%.

[0029] Using the same experimental method, sulfonamides 3b - 3o were respectively reacted with TMS-EBX iodide 4 to successfully prepare the corresponding alkynylamide intermediates 1b - 1o.

[0030]

[0031] Compound 1a: White solid, 99% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.58 (d, 2H, J = 8.3 Hz), 7.34 - 7.31 (m, 3H), 7.30 - 7.24 (m, 4H), 2.84 (s, 1H), 2.44 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 145.3, 138.4, 133.0, 129.7, 129.3, 128.6, 128.4, 126.4, 76.7, 59.1, 21.9.

[0032]

[0033] Compound 1b: White solid, 95% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.63 (d, 2H, J = 9.0 Hz), 7.35 - 7.31 (m, 3H), 7.28 - 7.25 (m, 2H), 6.94 (d, 2H, J = 9.0 Hz), 3.88 (s, 3H), 2.84 (s, 1H); 13 C NMR (100 MHz, CDCl 3)δ164.1, 138.5, 130.7, 129.3, 128.6, 127.6, 126.5, 114.2, 76.9, 59.1, 55.9.

[0034]

[0035] Compound 1c: White solid, 99% yield, 1 H NMR (400 MHz, CDCl 3 )δ7.52 (d, 2H, J = 8.0 Hz), 7.44 (t, 2H, J = 7.1 Hz), 7.39 - 7.35 (m, 1H), 3.12 (s, 3H), 2.96 (s, 1H); 13 C NMR (100 MHz, CDCl 3 )δ138.1, 129.7, 128.8, 125.8, 75.9, 59.9, 36.9.

[0036]

[0037] Compound 1d: White solid, 90% yield, 1 H NMR (400 MHz, CDCl 3 )δ7.59 (d, 2H, J = 8.4 Hz), 7.29 (d, 2H, J = 8.0 Hz), 7.14 - 7.09 (m, 4H), 2.81 (s, 1H), 2.45 (s, 3H), 2.34 (s, 3H); 13 C NMR (100 MHz, CDCl 3 )δ145.2, 138.8, 135.8, 133.1, 129.9, 129.7, 128.4, 126.4, 76.9, 58.7, 21.9, 21.3.

[0038]

[0039] Compound 1e: White solid, 99% yield, 1 H NMR (400 MHz, CDCl 3 )δ7.58 (d, 2H, J = 8.3 Hz), 7.29 (d, 2H, J = 8.0 Hz), 7.12 (d, 2H, J = 9.0 Hz), 6.82 (d, 2H, J = 9.0 Hz), 3.80 (s, 3H), 2.80 (s, 1H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3)δ159.7, 145.2, 133.0, 131.0, 129.7, 128.5, 128.1, 114.4, 77.1, 58.4, 55.7, 21.9.

[0040]

[0041] Compound 1f: White solid, 79% yield, 1 H NMR (400 MHz, CDCl 3 )δ7.58 (d, 2H, J = 8.3 Hz), 7.45 (d, 2H, J = 8.6 Hz), 7.30 (d, 2H, J = 8.1 Hz), 7.14 (d, 2H, J = 8.6 Hz), 2.85 (s, 1H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3 )δ145.6, 137.5, 132.7, 132.5, 129.8, 128.4, 127.9, 122.5, 76.2, 59.6, 21.9.

[0042]

[0043] Compound 1g: White solid, 80% yield, 1 H NMR (400 MHz, CDCl 3 )δ7.58 (d, 2H, J = 8.3 Hz), 7.30 (d, 4H, J = 8.9), 7.20 (d, 2H, J = 8.8 Hz), 2.85 (s, 1H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3 )δ145.5, 137.0, 134.4, 132.7, 129.8, 129.5, 128.4, 127.6, 76.2, 59.6, 21.9.

[0044]

[0045] Compound 1h: White solid, 68% yield, 1 H NMR (400 MHz, CDCl 3 )δ8.20 (d, 2H, J = 9.1 Hz), 7.61 (d, 2H, J = 8.4 Hz), 7.55 (d, 2H, J = 9.2 Hz), 7.31 (d, 2H, J = 8.2 Hz), 3.02 (s, 1H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3) δ 146.7, 146.1, 143.9, 132.7, 130.1, 128.3, 125.5, 124.7, 74.9, 61.7, 21.9; IR (neat) (cm -1 ) 3272 m, 2124 w, 1591 m, 1518 s, 1340 s, 1166 s, 682 s; HRMS (ESI): m / z calcd for C 15 H 13 N 2 O 4 S [M + H] + 317.0591, found 317.0594.

[0046]

[0047] Compound 1i: White solid, 93% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.60 (d, 2H, J = 8.3 Hz), 7.29 (d, 2H, J = 8.0 Hz), 7.20 (t, 1H, J = 7.8 Hz), 7.14 - 7.08 (m, 2H), 7.01 (d, 1H, J = 7.8 Hz), 2.82 (s, 1H), 2.45 (s, 3H), 2.32 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 145.2, 139.4, 138.3, 133.2, 129.7, 129.4, 129.0, 128.5, 127.2, 123.4, 76.8, 58.9, 21.9, 21.4.

[0048]

[0049] Compound 1j: White solid, 91% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.61 (d, 2H, J = 8.4 Hz), 7.29 (d, 2H, J = 8.1 Hz), 7.21 (t, 1H, J = 8.4 Hz), 6.87 - 6.81 (m, 3H), 3.76 (s, 3H), 2.84 (s, 1H), 2.44 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 160.1, 145.3, 139.4, 133.1, 129.9, 129.7, 128.4, 118.4, 114.5, 112.0, 76.9, 59.3, 55.6, 21.9.

[0050]

[0051] Compound 1k: White solid, 72% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.59 (d, 2H, J = 8.4 Hz), 7.46 - 7.43 (m, 1H), 7.42 (t, 1H, J = 2.0 Hz), 7.31 (d, 2H, J = 8.0 Hz), 7.25 - 7.24 (m, 1H), 7.23 - 7.19 (m, 1H), 2.89 (s, 1H), 2.46 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 145.6, 139.6, 132.8, 131.6, 130.5, 129.9, 129.1, 128.4, 124.9, 122.5, 76.0, 60.0, 21.9.

[0052]

[0053] Compound 1l: White solid, 72% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.60 (d, 2H, J = 8.4 Hz), 7.32 - 7.27 (m, 5H), 7.22 - 7.19 (m, 1H), 2.88 (s, 1H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 145.6, 139.5, 134.8, 132.8, 130.2, 129.9, 128.7, 128.4, 126.3, 124.4, 76.0, 60.0, 21.9; IR (neat) (cm -1 ) 3290w, 2131w, 1587m, 1371s, 1166s, 1085m, 670s; HRMS (ESI): m / z calcd for C 15 H 13 ClNO 2 S [M + H] + 306.0350, found 306.0346.

[0054]

[0055] Compound 1m: White solid, 81% yield, 1 H NMR (400 MHz, CDCl 3) δ 7.60 (d, 2H, J = 8.3 Hz), 7.30 (d, 3H, J = 8.3 Hz), 7.12 - 7.10 (m, 1H), 7.05 - 7.00 (m, 2H), 2.89 (s, 1H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 162.6 (C - F, 1 J C-F = 246.7 Hz), 145.6, 139.8 (C - F, 3 J C-F = 9.8 Hz), 132.8, 130.3 (C - F, 3 J C-F = 8.9 Hz), 129.9, 128.4, 121.7 (C - F, 4 J C-F = 3.3 Hz), 115.5 (C - F, 2 J C-F = 20.8 Hz), 113.5 (C - F, 2 J C-F = 24.4 Hz), 76.0, 60.0, 21.9.

[0056]

[0057] Compound 1n: White solid, 61% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 8.18 (ddd, 1H, J = 8.2 Hz, 2.2 Hz, 1.0 Hz), 8.11 (t, 1H, J = 2.2 Hz), 7.76 (ddd, 1H, J = 8.1, 2.2, 1.0 Hz), 7.60 (d, 2H, J = 8.4 Hz), 7.56 (t, 1H, J = 8.2 Hz), 7.32 (d, 2H, J = 8.0 Hz), 2.96 (s, 1H), 2.46 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 148.6, 146.1, 139.8, 132.5, 132.0, 130.2, 130.1, 128.4, 123.0, 120.6, 75.3, 61.0, 22.0; IR (neat) (cm -1 ) 3296w, 2129w, 1527s, 1348s, 1165s, 1077m, 666s; HRMS (ESI): m / z calcd for C 15 H 13 N 2 O4 S[M+H] + 317.0591, found 317.0593.

[0058]

[0059] Compound 1o: White solid, 86% yield, 1 H NMR (400 MHz, CDCl 3 ) δ 7.62 (d, 2H, J = 8.3 Hz), 7.30 (d, 2H, J = 8.1 Hz), 6.95 (s, 1H), 6.85 (s, 2H), 2.81 (s, 1H), 2.45 (s, 3H), 2.27 (s, 6H); 13 C NMR (100 MHz, CDCl 3 ) δ 145.1, 139.1, 138.2, 133.3, 130.4, 129.6, 128.5, 124.2, 77.0, 58.8, 21.9, 21.3.

[0060] Example 2: Synthesis of target compounds 2a - 2o

[0061] Specific note: In the compound structure, Ts represents p - toluenesulfonyl, Mbs represents p - methoxybenzenesulfonyl, and Ms represents methylsulfonyl.

[0062]

[0063] Taking the synthesis step of compound 2a as an example: Add propargylamide 1a (81.4 mg, 0.3 mmol) to a dry reaction flask. After three nitrogen replacements, heat it in an oil bath to 100 °C until it melts completely. Maintain this temperature for 2.0 h, and monitor the reaction progress by TLC during this period. After the reaction is completed, cool it to room temperature. The crude product is purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3.5:1) to obtain white solid 2a (70.8 mg, 0.131 mmol), with a separation yield of 87%.

[0064] Expanded synthesis: Using the same experimental method, subject the propargylamide intermediates 1b - 1o prepared in Example 1 to cyclization reactions respectively, and successfully prepare the corresponding target compounds 2b - 2o.

[0065]

[0066] Compound 2a: White solid, 87% yield, R f = 0.25 [petroleum ether / ethyl acetate (2:1)]; mp = 86–89 °C; 1 H NMR (400 MHz, CDCl 3) δ 7.79 (d, 2H, J = 8.3 Hz), 7.48 (d, 2H, J = 8.4 Hz), 7.46 - 7.38 (m, 3H), 7.33 - 7.24 (m, 6H), 7.18 - 7.08 (m, 3H), 6.76 - 6.73 (m, 2H), 5.94 (s, 1H), 4.91 (s, 1H), 2.45 (s, 3H), 2.42 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 153.1, 152.0, 148.9, 146.0, 145.5, 135.7, 133.1, 132.9, 130.2, 130.1, 130.02, 129.97, 129.5, 129.4, 129.1, 128.4, 124.8, 121.7, 118.6, 76.0, 21.94, 21.89; IR (neat) (cm -1 ) 1769 w, 1695 w, 1552 s, 1486 m, 1320 m, 1173 s, 694 s; HRMS (ESI): m / z calcd for C 30 H 27 N 2 O 4 S 2 [M + H] + 543.1407, found 543.1411.

[0067]

[0068] Compound 2b: White solid, 82% yield, R f = 0.12 [petroleum ether / ethyl acetate (2:1)]; mp = 94–97 °C; 1 H NMR (400 MHz, CDCl 3 ) δ 7.84 (d, 2H, J = 8.9 Hz), 7.52 (d, 2H, J = 9.0 Hz), 7.46 - 7.38 (m, 3H), 7.28 (t, 2H, J = 7.7 Hz), 7.17 - 7.09 (m, 3H), 6.98 (d, 2H, J = 8.9 Hz), 6.89 (d, 2H, J = 9.0 Hz), 6.78 (d, 2H, J = 7.3 Hz), 5.93 (s, 1H), 4.91 (s, 1H), 3.87 (s, 3H), 3.84 (s, 3H); 13 C NMR (100 MHz, CDCl 3) δ 164.5, 164.3, 153.3, 152.2, 148.9, 135.7, 132.2, 130.6, 130.11, 130.07, 129.3, 129.1, 127.3, 124.8, 121.8, 118.2, 114.5, 114.0, 76.1, 55.9, 55.8, where there is a carbon signal overlap at 129.1 ppm; IR (neat) (cm -1 ) 1697w, 1592m, 1551s, 1260s, 1136s, 1083s, 832s; HRMS (ESI): m / z calcd for C 30 H 27 N 2 O 6 S 2 [M + H] + 575.1305, found 575.1303.

[0069]

[0070] Compound 2c: White solid, 71% yield, R f = 0.24 [petroleum ether / ethyl acetate (2:1)]; mp = 162–164 °C; 1 1H NMR (400 MHz, CDCl 3 ) δ 7.52 - 7.45 (m, 5H), 7.30 - 7.26 (m, 2H), 7.10 (t, 1H, J = 7.4 Hz), 6.96 (d, 2H, J = 7.4 Hz), 5.53 (s, 1H), 5.41 (s, 1H), 3.35 (s, 3H), 3.28 (s, 3H); 13 13C NMR (100 MHz, CDCl 3 ) δ 153.0, 152.1, 148.5, 136.1, 130.5, 130.3, 129.5, 129.2, 125.1, 121.7, 117.5, 74.9, 42.9, 40.0; IR (neat) (cm -1 ) 1699m, 1591w, 1564s, 1483m, 1299s, 1124m, 760s; HRMS (ESI): m / z calcd for C 18 H 19 N 2 O 4 S 2 [M + H] + 391.0781, found 391.0780.

[0071]

[0072] Compound 2d: White solid, 81% yield, R f = 0.37 [petroleum ether / ethyl acetate (2:1)]; mp = 100–101 °C; 1 H NMR (400 MHz, CDCl 3 ) δ 7.79 (d, 2H, J = 8.3 Hz), 7.48 (d, 2H, J = 8.4 Hz), 7.31 (d, 2H, J = 8.0 Hz), 7.25 (d, 2H, J = 8.0 Hz), 7.19 (d, 2H, J = 8.3 Hz), 7.08 (d, 2H, J = 8.0 Hz), 7.02 (d, 2H, J = 8.1 Hz), 6.64 (d, 2H, J = 8.2 Hz), 5.93 (s, 1H), 4.87 (s, 1H), 2.45 (s, 3H), 2.42 (s, 3H), 2.40 (s, 3H), 2.32 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 152.6, 151.9, 146.4, 145.9, 145.4, 140.4, 134.5, 133.1, 133.0, 132.7, 130.1, 130.0, 129.9, 129.8, 129.7, 129.4, 128.4, 121.7, 118.6, 76.0, 21.94, 21.88, 21.5, 21.1; IR (neat) (cm -1 ) 1695 w, 1554 s, 1450 w, 1320 m, 1172 s, 1137 s, 667 s; HRMS (ESI): m / z calcd for C 32 H 31 N 2 O 4 S 2 [M+H] + 571.1720, found 571.1716.

[0073]

[0074] Compound 2e: White solid, 76% yield, R f = 0.13 [petroleum ether / ethyl acetate (2:1)]; mp = 95–97 °C; 1 H NMR (400 MHz, CDCl 3)δ 7.79 (d, 2H, J = 8.2 Hz), 7.48 (d, 2H, J = 8.4 Hz), 7.31 (d, 2H, J = 8.2 Hz), 7.26 (d, 2H, J = 9.6 Hz), 7.05 (s, 2H), 6.87 (d, 2H, J = 9.0 Hz), 6.82 (d, 2H, J = 8.9 Hz), 6.71 (d, 2H, J = 8.9 Hz), 5.96 (s, 1H), 4.86 (s, 1H), 3.84 (s, 3H), 3.79 (s, 3H), 2.45 (s, 3H), 2.42 (s, 3H); 13 C NMR (100 MHz, CDCl 3 )δ 160.6, 157.1, 151.99, 151.95, 145.9, 145.4, 142.1, 133.1, 132.8, 131.4, 130.0, 129.9, 129.4, 128.4, 127.9, 123.0, 118.5, 114.4, 114.3, 76.0, 55.63, 55.58, 21.93, 21.87; IR (neat) (cm -1 ) 1768 w, 1555 s, 1501 s, 1242 s, 1170 s, 1082 m, 668 s; HRMS (ESI): m / z calcd for C 32 H 31 N 2 O 6 S 2 [M + H] + 603.1618, found 603.1614.

[0075]

[0076] Compound 2f: White solid, 85% yield, R f = 0.50 [petroleum ether / ethyl acetate (2:1)]; mp = 99–102 °C; 1 1H NMR (400 MHz, CDCl 3 )δ 7.75 (d, 2H, J = 8.3 Hz), 7.54 - 7.49 (m, 4H), 7.39 (d, 2H, J = 8.6 Hz), 7.33 - 7.28 (m, 4H), 7.02 (d, 2H, J = 8.1 Hz), 6.61 (d, 2H, J = 8.6 Hz), 5.92 (s, 1H), 4.87 (s, 1H), 2.46 (s, 3H), 2.44 (s, 3H); 13 C NMR (100 MHz, CDCl 3) δ 153.4, 151.9, 147.9, 146.4, 145.7, 134.6, 132.9, 132.8, 132.7, 132.2, 131.6, 130.2, 129.9, 129.6, 128.4, 124.7, 123.4, 118.4, 118.1, 75.8, 21.99, 21.95; IR (neat) (cm -1 ) 1695w, 1551s, 1399w, 1373m, 1284m, 1174s, 662s; HRMS (ESI): m / z calcd for C 30 H 25 Br 2 N 2 O 4 S 2 [M + H] + 698.9617, found 698.9615.

[0077]

[0078] Compound 2g: White solid, 84% yield, R f = 0.46 [petroleum ether / ethyl acetate (2:1)]; mp = 104–105 °C; 1 H NMR (400 MHz, CDCl 3 ) δ 7.75 (d, 2H, J = 8.3 Hz), 7.50 (d, 2H, J = 8.3 Hz), 7.37 - 7.27 (m, 6H), 7.24 (d, 2H, J = 8.6 Hz), 7.12 - 7.06 (m, 2H), 6.67 (d, 2H, J = 8.6 Hz), 5.93 (s, 1H), 4.88 (s, 1H), 2.46 (s, 3H), 2.43 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 153.4, 151.9, 147.4, 146.4, 145.7, 136.5, 134.1, 132.9, 132.8, 131.3, 130.3, 130.2, 129.9, 129.7, 129.6, 129.2, 128.4, 123.0, 118.4, 75.8, 22.0, 21.9; IR (neat) (cm -1 ) 1690w, 1554s, 1374m, 1138s, 1083s, 1016m, 666s; HRMS (ESI): m / z calcd for C 30 H 25 Cl 2 N 2 O4 S 2 [M+H] + 611.0627, found 611.0630.

[0079]

[0080] Compound 2h: white solid, 52% yield, R f = 0.25 [petroleum ether / ethyl acetate (2:1)]; mp = 174–176 °C; 1 H NMR (400 MHz, CDCl 3 ) δ 8.27 (d, 2H, J = 8.6 Hz), 8.17 (d, 2H, J = 8.5 Hz), 7.73 (d, 2H, J = 8.0 Hz), 7.52 (d, 2H, J = 8.0 Hz), 7.39 - 7.31 (m, 6H), 6.82 (d, 2H, J = 8.8 Hz), 5.99 (s, 1H), 4.93 (s, 1H), 2.48 (s, 3H), 2.46 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 154.8, 154.6, 152.1, 148.4, 147.1, 146.2, 144.9, 141.0, 132.9, 132.6, 131.0, 130.5, 129.8, 128.3, 125.2, 124.8, 121.9, 118.2, 75.6, 22.0, where two carbon signals overlap at 129.8 ppm and 22.0 ppm; IR (neat) (cm -1 ) 1700 w, 1555 s, 1339 s, 1170 m, 1081 m, 1016 w, 662 s; HRMS (ESI): m / z calcd for C 30 H 25 N 4 O 8 S 2 [M+H] + 633.1108, found 633.1110.

[0081]

[0082] Compound 2i: white solid, 91% yield, R f = 0.40 [petroleum ether / ethyl acetate (2:1)]; mp = 77–78 °C; 1 H NMR (400 MHz, CDCl 3) δ 7.79 (d, 2H, J = 8.3 Hz), 7.50 (d, 2H, J = 8.4 Hz), 7.32 (d, 2H, J = 7.9 Hz), 7.27 - 7.24 (m, 4H), 7.16 (t, 1H, J = 7.9 Hz), 6.98 - 6.89 (m, 3H), 6.54 - 6.51 (m, 2H), 5.90 (s, 1H), 4.90 (s, 1H), 2.46 (s, 3H), 2.43 (s, 3H), 2.35 (s, 3H), 2.31 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 152.9, 151.9, 148.9, 145.9, 145.4, 139.5, 138.9, 135.5, 133.2, 133.0, 130.9, 130.7, 130.1, 129.9, 129.4, 129.0, 128.9, 128.5, 127.0, 125.6, 122.4, 118.7, 118.6, 76.1, 21.93, 21.88, 21.6, 21.5; IR (neat) (cm -1 ) 1769 w, 1595 w, 1551 s, 1370 m, 1172 s, 1136 s, 670 s; HRMS (ESI): m / z calcd for C 32 H 31 N 2 O 4 S 2 [M + H] + 571.1720, found 571.1718.

[0083]

[0084] Compound 2j: White solid, 90% yield, R f = 0.19 [petroleum ether / ethyl acetate (2:1)]; mp = 76–77 °C; 1 H NMR (400 MHz, CDCl 3 ) δ 7.79 (d, 2H, J = 8.3 Hz), 7.53 (d, 2H, J = 8.4 Hz), 7.32 (d, 2H, J = 8.0 Hz), 7.29 - 7.25 (m, 3H), 7.17 (t, 1H, J = 8.4 Hz), 6.99 (dd, 1H, J = 8.0, 2.1 Hz), 6.74 (s, 1H), 6.68 - 6.65 (m, 2H), 6.31 - 6.29 (m, 2H), 5.92 (s, 1H), 4.94 (s, 1H), 3.78 (s, 3H), 3.76 (s, 3H), 2.45 (s, 3H), 2.42 (s, 3H);13 C NMR (100 MHz, CDCl 3 ) δ 160.3, 160.1, 153.3, 152.0, 150.3, 146.0, 145.4, 136.5, 133.2, 133.1, 130.0, 129.9, 129.81, 129.76, 129.5, 128.5, 122.0, 118.7, 116.0, 113.8, 110.5, 107.6, 76.1, 55.7, 55.4, 21.92, 21.88, where there is a carbon signal overlap at 129.9 ppm; IR (neat) (cm -1 ) 1700 w, 1551 s, 1449 w, 1370 m, 1135 s, 1040 m, 673 s; HRMS (ESI): m / z calcd for C 32 H 31 N 2 O 6 S 2 [M + H] + 603.1618, found 603.1622.

[0085]

[0086] Compound 2k: White solid, 91% yield, R f = 0.42 [petroleum ether / ethyl acetate (2:1)]; mp = 88–89 °C; 1 H NMR (400 MHz, CDCl 3 ) δ 7.75 (d, 2H, J = 8.3 Hz), 7.60 (d, 1H, J = 8.1 Hz), 7.52 (d, 2H, J = 7.8 Hz), 7.35 - 7.29 (m, 5H), 7.23 (d, 2H, J = 8.1 Hz), 7.14 (t, 2H, J = 7.8 Hz), 6.72 (t, 1H, J = 1.8 Hz), 6.70 (d, 1H, J = 7.8 Hz), 5.89 (s, 1H), 4.91 (s, 1H), 2.48 (s, 3H), 2.45 (s, 3H); 13 C NMR (100 MHz, CDCl 3)δ 153.8, 151.9, 150.3, 146.5, 145.8, 136.7, 133.4, 132.9, 132.8, 132.7, 130.6, 130.5, 130.2, 129.9, 129.6, 128.9, 128.4, 127.7, 124.4, 122.6, 122.5, 120.4, 118.3, 75.8, 22.0, 21.9; IR (neat) (cm -1 )1696 w, 1547 s, 1373 m, 1288 s, 1175 s, 1019 w, 664 s; HRMS (ESI): m / z calcd for C 30 H 25 Br 2 N 2 O 4 S 2 [M + H] + 698.9617, found 698.9614.

[0087]

[0088] Compound 2l: White solid, 91% yield, R f = 0.35 [petroleum ether / ethyl acetate (2:1)]; mp = 82–83 °C; 1 1H NMR (400 MHz, CDCl 3 ) δ 7.75 (d, 2H, J = 8.3 Hz), 7.52 (d, 2H, J = 8.2 Hz), 7.45 (d, 1H, J = 8.1 Hz), 7.37 - 7.29 (m, 5H), 7.20 (t, 1H, J = 8.0 Hz), 7.13 - 7.07 (m, 3H), 6.64 (d, 1H, J = 7.8 Hz), 6.60 (t, 1H, J = 2.0 Hz), 5.91 (s, 1H), 4.91 (s, 1H), 2.47 (s, 3H), 2.44 (s, 3H); 13 13C NMR (100 MHz, CDCl 3 ) δ 153.8, 151.9, 150.1, 146.5, 145.8, 136.6, 134.9, 134.6, 132.9, 132.8, 130.5, 130.3, 130.17, 130.16, 130.13, 129.9, 129.6, 128.4, 124.8, 121.6, 120.0, 118.4, 75.9, 21.95, 21.92, where there is a carbon signal overlap at 130.16 ppm; IR (neat) (cm -1)1696w,1550s,1288m,1175m,1081m,1021w,667s; HRMS(ESI): m / z calcd for C 30 H 25 Cl 2 N 2 O 4 S 2 [M+H] + 611.0627, found 611.0628.

[0089]

[0090] Compound 2m: White solid, 88% yield, R f = 0.37 [petroleum ether / ethyl acetate (2:1)]; mp = 81–82 °C; 1 H NMR(400 MHz, CDCl 3 ) δ 7.76 (d, 2H, J = 8.0 Hz), 7.53 (d, 2H, J = 8.0 Hz), 7.41 - 7.29 (m, 5H), 7.24 - 7.11 (m, 2H), 6.97 - 6.92 (m, 2H), 6.82 (td, 1H, J = 8.4, 2.6 Hz), 6.54 (d, 1H, J = 8.0 Hz), 6.41 (dt, 1H, J = 10.0, 2.1 Hz), 5.94 (s, 1H), 4.92 (s, 1H), 2.47 (s, 3H), 2.45 (s, 3H); 13 C NMR(100 MHz, CDCl 3 ) δ 163.2 (C-F, 1 J C-F = 244.8 Hz), 162.5 (C-F, 1 J C-F = 248.2 Hz), 153.7, 151.9, 150.6 (C-F, 3 J C-F = 9.0 Hz), 146.4, 145.8, 136.8 (C-F, 3 J C-F = 9.8 Hz), 133.0, 132.9, 130.4 (C-F, 3 J C-F = 8.8 Hz), 130.3 (C-F, 3 J C-F = 9.3 Hz), 130.1, 129.9, 129.6, 128.3, 125.7 (C-F, 4 J C-F= 2.5 Hz), 118.5, 117.6 (C-F, 2 J C-F = 23.3 Hz), 117.5 (C-F, 2 J C-F = 20.8 Hz), 117.4 (C-F, 4 J C-F = 2.8 Hz), 111.6 (C-F, 2 J C-F = 21.1 Hz), 108.9 (C-F, 2 J C-F = 22.4 Hz), 75.9, 21.9, where there is a carbon signal overlap at 21.9 ppm; 19 F NMR(376 MHz, CDCl 3 ) δ -109.9, -112.4; IR(neat)(cm -1 ) 1702 w, 1552 s, 1480 m, 1373 m, 1173 s, 1024 w, 670 s; HRMS(ESI): m / z calcd for C 30 H 25 F 2 N 2 O 4 S 2 [M + H] + 579.1218, found 579.1216.

[0091]

[0092] Compound 2n: White solid, 71% yield, R f = 0.17 [petroleum ether / ethyl acetate (2:1)]; mp = 211–213 °C; 1 H NMR(400 MHz, CDCl 3 ) δ 8.37 - 8.35 (m, 1H), 7.98 (d, 1H, J = 8.1 Hz), 7.90 (s, 1H), 7.74 (d, 2H, J = 8.0 Hz), 7.69 (d, 2H, J = 5.6 Hz), 7.53 (d, 2H, J = 8.2 Hz), 7.47 (t, 1H, J = 8.0 Hz), 7.40 - 7.33 (m, 5H), 7.13 (d, 1H, J = 8.6 Hz), 5.97 (s, 1H), 4.92 (s, 1H), 2.49 (s, 3H), 2.47 (s, 3H); 13 C NMR(100 MHz, CDCl 3)δ 154.7, 152.0, 150.0, 148.9, 148.4, 147.2, 146.4, 136.8, 136.6, 132.52, 132.48, 130.5, 130.5, 130.1, 129.9, 129.8, 128.3, 128.0, 125.2, 124.8, 119.7, 118.0, 116.1, 75.7, 22.01, 21.99; IR (neat) (cm -1 ) 1706w, 1563s, 1522s, 1347s, 1174m, 1079m, 672s; HRMS (ESI): m / z calcd for C 30 H 25 N 4 O 8 S 2 [M + H] + 633.1108, found 633.1111.

[0093]

[0094] Compound 2o: White solid, 87% yield, R f = 0.50 [petroleum ether / ethyl acetate (2:1)]; mp = 132–134 °C; 1 1H NMR (400 MHz, CDCl 3 ) δ 7.79 (d, 2H, J = 8.2 Hz), 7.51 (d, 2H, J = 8.4 Hz), 7.31 (d, 2H, J = 8.0 Hz), 7.26 (d, 2H, J = 8.2 Hz), 7.06 (s, 1H), 6.78 - 6.72 (m, 3H), 6.30 (s, 2H), 5.88 (s, 1H), 4.89 (s, 1H), 2.46 (s, 3H), 2.43 (s, 3H), 2.29 (s, 6H), 2.26 (s, 6H); 13 13C NMR (100 MHz, CDCl 3 ) δ 152.8, 151.9, 148.9, 145.8, 145.2, 139.1, 138.7, 135.3, 133.3, 133.0, 131.8, 130.1, 129.8, 129.4, 128.6, 127.6, 126.4, 119.4, 118.8, 76.1, 21.92, 21.88, 21.5, 21.4; IR (neat) (cm -1 ) 2920w, 1594w, 1553s, 1318m, 1147s, 1082m, 683s; HRMS (ESI): m / z calcd for C 34H 35 N 2 O 4 S 2 [M+H] + 599.2033, found 599.2031.

[0095] Example 4: Study on the antiviral activity of the representative compounds synthesized in Example 2 of the present invention

[0096] The experimental reagents, experimental instruments and their sources used in the study of the antiviral activity of Compounds 2a - 2o are shown in Tables 1 and 2.

[0097] Table 1 Experimental reagents and manufacturers

[0098]

[0099] Table 2 Experimental instruments and manufacturers

[0100]

[0101]

[0102] Detection of cytotoxicity and antiviral activity of Compounds 2a - 2o

[0103] Hep - 2 cells in good growth state were inoculated into 96 - well cell culture plates at a density of 1.5×10 4 cells / well. After 24 h of adhesion, the old culture medium was discarded and the cells were washed 3 times with PBS. The compounds were dissolved in DMSO to prepare a stock solution of 20 mM. Then, it was serially diluted two - fold with the virus maintenance medium (from 800 μM to 6.25 μM, a total of 8 gradients). 200 μL of the treatment solution was added to each well, and at the same time, blank control replicates were set, with 200 μL of the virus maintenance medium added to each well, and 6 replicates for each concentration. After 72 h of compound treatment, 20 μL of MTT working solution was added to each well, and the mixture was incubated at 37 °C in the dark for 4 h, during which purple formazan crystals were formed. After the incubation, the original solution was discarded, and the supernatant was carefully aspirated. 160 μL of DMSO was added. The plate was shaken on a horizontal shaker for 15 min to ensure complete dissolution of the crystals. The OD 492nm was read using an enzyme - linked immunosorbent assay reader. The formula for calculating the cell survival rate is as follows: Cell survival rate = (OD 492nm of the compound group / OD 492nm of the normal group) × 100%. The half - cytotoxic concentration (CC 50 ) of the compounds was calculated using IBM SPSS 27 software, and the results are shown in Table 3.

[0104] Table 3 Detection results of cytotoxicity and antiviral activity of Compounds 2a - 2o

[0105]

[0106]

[0107] Note: a Indicates the 50% cytotoxic concentration against Hep-2 cells, b Indicates the 50% effective concentration against HSV-1, c Indicates the therapeutic index SI, d Indicates no anti-HSV-1 activity between 0.625 μM and 100 μM.

[0108] Hep-2 cells in good growth state were seeded in a 96-well cell culture plate at a density of 1.5×10 4 cells / well. After 24 h of adherence, the old medium was discarded and the cells were washed 3 times with PBS. The virus stock solution was diluted to 100 TCID 50 with the virus adsorption solution, and 100 μL was added to each well. At the same time, blank control wells were set, adding only 100 μL of the virus adsorption solution, with 6 replicates for each concentration. The treated 96-well plate was placed in an incubator, taken out and shaken every 15 min for a total of 4 times, and then left to stand in the incubator for 1 h to ensure sufficient virus adsorption. The adsorption solution was removed, and the unbound virus was removed with PBS. The compound was dissolved in DMSO to prepare a 20 mM stock solution, and then serially diluted two-fold with the virus maintenance solution (from 400 μM to 3.125 μM, a total of 8 gradients). 200 μL of the corresponding dilution sample was supplemented to each well. At the same time, virus and blank control replicates were set, adding only 200 μL of the virus maintenance solution, with 6 replicates for each concentration. After 72 h, the OD 492nm was read with an enzyme-linked immunosorbent assay reader. The virus inhibition rate was calculated using the following formula: Virus inhibition rate = (OD of the compound group 492nm - OD of the virus group 492nm ) / (OD of the normal group 492nm - OD of the virus group 492nm ) × 100%. The half-maximal effective concentration (EC 50 ) of the compound was calculated using IBM SPSS 27 software, and the therapeutic index (SI) was calculated according to the following formula: SI = CC 50 / EC 50 . The specific results are shown in Table 3.

[0109] In this invention, the internationally standardized MTT assay well-known to those skilled in the art was used, and the Hep-2 cell line was used as an in vitro model to systematically evaluate the anti-HSV-1 / KOS strain activity and cytotoxicity of aminocyclobutene compounds 2a-2o. The experimental results showed that compounds 2k (EC 50 = 3.80 ± 0.98 μM) and 2n (EC 50 = 49.58 ± 3.47 μM) both showed significant antiviral activity. Notably, the half-maximal effective concentration of compound 2k was lower than that of acyclovir, the first-line drug in clinical use (EC50 = 9.62 ± 3.06 μM), a 60.5% reduction, showing a significant antiviral activity advantage. In terms of drug safety evaluation, compound 2k exhibited excellent biocompatibility characteristics, with its half cytotoxic concentration (CC 50 ) as high as 390.16 ± 10.79 μM, and the calculated therapeutic index (SI = 102.67) reached 3.5 times that of acyclovir (SI = 29.45). This excellent activity-safety balance characteristic makes compound 2k a candidate molecule with great development value. Based on the above experimental results, the aminocyclobutene compounds of the present invention have good application prospects in the preparation and research of anti-HSV-1 drugs.

Claims

1. The pharmaceutical application of the aminocyclobutene compound with the following general structural formula, characterized in that: It is used as an active ingredient in the preparation of drugs against herpes simplex virus type 1: EWG is selected from sulfonyl, aryl-substituted sulfonyl or C1-3 alkyl-substituted sulfonyl; the aryl is selected from phenyl substituted by methyl or methoxy; R is selected from hydrogen, C1-3 alkyl, C1-3 alkoxy, halogen or nitro, and R is monosubstituted at the ortho / meta / para position of the benzene ring.

2. The pharmaceutical use of the aminocyclobutene compound according to claim 1, characterized in that: EWG is selected from p-toluenesulfonyl, p-methoxybenzenesulfonyl, methylsulfonyl; and R is selected from halogen or nitro.

3. The pharmaceutical use of the aminocyclobutene compound as claimed in claim 2, characterized in that: Selected from the following compounds:

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