Ibuprofen triazole thiol derivatives and their applications in the preparation of novel coronavirus inhibitors

Abstract

The present invention relates to ibuprofen triazole thiol derivatives or ibuprofen triazole thione derivatives shown in structural formula I or II, their preparation methods, pharmaceutical compositions, and their use in the preparation of novel coronavirus 3CL protease inhibitors. Ibuprofen triazole thione is a tautomer of ibuprofen triazole thiol derivatives. Wherein R is selected from: hydrogen, deuterium, C1-C2 alkyl, C3-C7 straight chain or C3-C7 branched alkyl; Z is selected from: hydrogen, 4-fluoro, 4-hydroxy, 2-(2,6-dichloroanilino), 4-isobutyl, 3-fluoro-4-phenyl, 3-phenoxy, 3-benzoyl, 4-(2-methylallylamino) or 4-(2-oxocyclopentylmethyl); Y 1 、Y 2 and Y 4 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy; Y 3 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, hydroxy, ethoxy, nitro, amino, methylamino, dimethylamino, acetylamino, fluorine, chlorine, bromine or iodine; Y 5 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine or iodine.

Description

Technical Field

[0001] The present invention relates to a new class of compounds, their preparation methods and applications, specifically ibuprofen triazole thiol derivatives or ibuprofen triazole thione derivatives and their applications in the preparation of novel coronavirus 3CL protease inhibitors. Background Art

[0002] Guo Changbin et al. [Acta Chimica Sinica, 2005, 63(09):841-848] described the use of the Autodock program to simulate the conformation of ibuprofen binding to COX-1 and COX-2, and compared it with the structure of the COX-2 selective inhibitor SC-558 and COX-2 crystal complex. They found that ibuprofen lacked a structural fragment that occupied the side pocket of COX-2, so it had no selectivity for the two isozymes. 2-(3-substituted benzamido-4-isobutylphenyl) propionic acid, a substituted benzamide group, was introduced at the 3-position of the benzene ring of ibuprofen to occupy the side pocket of COX-2, increase the binding effect on COX-2, and obtain a selective inhibitor for COX-2.

[0003]

[0004] Shanbhag et al. [J Pharmaceutical Sciences, 1992, 81(2):149–154] described that the gastrointestinal irritation of ibuprofen is mainly caused by the carboxyl group on the ibuprofen molecule, and that ibuprofen was made into a prodrug to improve patient compliance. Ibuprofen was prepared as eugenol ester, which does not have the volatility and irritation of eugenol and can also reduce the gastrointestinal irritation of ibuprofen [Journal of Shenyang Pharmaceutical University, 2006, 23(2), 70-73; Eur J Pharmaceutical Sciences, 2002, 17(3), 121-130]. ​​Bhat et al. [Joseph J. 1989, 61(4):134-136] described the synthesis of a series of ibuprofen derivatives by esterification of ibuprofen with ROH, among which the sulfonamide derivatives of ibuprofen showed activity against Canadian mold.

[0005] Chinese invention patents [ZL201010143067.2; ZL201210106644.X] describe the anti-inflammatory and antidepressant activities of ibuprofen-2-arylmorpholine ethyl ester (1):

[0006]

[0007] 4-(Arylmethyleneamino)-3-alkyl-1H-1,2,4-triazole-5(4H)-thiones (2) and their biological activities are summarized as follows:

[0008]

[0009]

[0010] Siddiqui et al. [Oriental Journal of Chemistry, 2005, 21(2): 317-322] described that 3-phenyl-4-benzylideneamino-1,2,4-triazolethione compounds (3) have certain analgesic and anti-inflammatory activities. When R = 4-Cl or 4-NO2, the analgesic and anti-inflammatory activities are better than the control drug sodium diclofenac.

[0011]

[0012] Gowda et al. [European Journal of Medicinal Chemistry, 2011, 46: 4100-4106] disclosed a triazolethione-containing benzothiazinone derivative 4, which was tested for its in vivo anti-inflammatory activity at a concentration of 20 mg / kg. It was found that when X = O and Ar = 4-HOC6H4 or 4-NO2C6H4, its anti-inflammatory effect was slightly inferior to that of indomethacin.

[0013]

[0014] Lu Wenting et al. [Chemical Bulletin, 2012, 75(4): 361-364] described the fungicidal activity of 5-benzyl-1,2,4-triazole-3-thiol (5). 3-Benzyl-1H-1,2,4-triazole-5(4H)-thione is a tautomer of 5-benzyl-1,2,4-triazole-3-thiol.

[0015]

[0016] et al. [E-Journal of Chemistry, 2010, 7(S1):S458-S464] described the fungicidal activity of 3-(1-phenylethyl)-1,2,4-triazole-5-thione (6). 3-(1-phenylethyl)-1H-1,2,4-triazole-5(4H)-thione is a tautomer of 5-(1-phenylethyl)-1,2,4-triazole-3-thiol.

[0017]

[0018] Chinese invention patents [CN109053606A, published on December 21, 2018; CN109053607A, published on December 21, 2018] describe the use of 4-(4-hydroxyphenylmethyleneamino)-1H-1,2,4-triazole-5(4H)-thiones [7,8: J. Heterocyclic Chem., 2019, 56, 2192] in the preparation of influenza virus neuraminidase inhibitors. 3-Alkyl-1H-1,2,4-triazole-5(4H)-thiones are tautomers of 5-alkyl-1,2,4-triazole-3-thiols.

[0019]

[0020] Among them, R 1 is selected from the group consisting of hydrogen, C1-C2 alkyl, C3-C7 straight chain or C3-C7 branched alkyl; phenyl, 4-fluorophenyl; X is selected from the group consisting of H, methyl, ethyl, amino or hydroxy; R is selected from the group consisting of hydrogen, methoxy, ethoxy, C3-C4 straight chain alkoxy or C3-C4 branched alkoxy; Y is selected from the group consisting of phenyl, 4-hydroxyphenyl or 4-methoxyphenyl, fluoromethyl, difluoromethyl or trifluoromethyl.

[0021] Sujith et al. [European Journal of Medicinal Chemistry, 2009, 44: 3697–3702] used ibuprofen as a raw material to synthesize 5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol (9), where Y = H, 4-Cl, 4-Br, 4-CH3, 4-NO2 and 2,6-Cl2. The in vivo anti-inflammatory activity showed that the introduction of 1,2,4-triazole thiol into the ibuprofen molecule could enhance its anti-inflammatory activity. When R = 4-Cl, the anti-inflammatory activity was the best, which was better than ibuprofen. Martin et al. [International Journal of Pharmaceutical Education and Research, 2019, 1(2): 47-51] reported that 5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol 9, where Y = H, 4-Cl, 4-OH, 4-OCH3, 4-OH-3-OCH3, 4-N(CH3)2 and 3,4,5-(OCH3)3, had certain antibacterial activity. Dhall et al. [J. Heterocyclic Chem., 2018, 55: 2859-2869] also reported the synthesis of 5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol 9, where Y = H, 2-Cl, 2-Br, 2-OCH3, 3-NO2, 4-Cl, 4-Br, 4-OH, 4-OCH3 and 4-N(CH3)2. 3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione is a tautomer of 5-(1-(4-isobutylphenyl)ethyl))-1,2,4-triazole-3-thiol.

[0022]

[0023] Naser et al. [International Journal of Pharmaceutical Sciences and Research, 2017, 8(4):1598-1605] reported the synthesis of 5-(1-(3-fluoro-4-biphenyl)ethyl)-1,2,4-triazole-3-thiol (10). 3-(1-(3-fluoro-4-biphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione is a tautomer of 5-(1-(3-fluoro-4-biphenyl)ethyl)-1,2,4-triazole-3-thiol (10).

[0024] Summary of the Invention

[0025] The technical problem solved by the present invention is to provide ibuprofen triazole thiol derivatives or ibuprofen triazole thione derivatives, their preparation methods, pharmaceutical compositions and uses in the preparation of novel coronavirus 3CL protease inhibitors.

[0026] In order to solve the technical problems of the present invention, the present invention provides the following technical solutions:

[0027] The first aspect of the technical solution of the present invention provides a class of ibuprofen triazole thiol derivatives or ibuprofen triazole thione derivatives as shown in structural formula I or II:

[0028]

[0029] Ibuprofen triazolethione derivatives are tautomers of the corresponding ibuprofen triazolethiol derivatives, the same below.

[0030] Wherein, R is selected from: hydrogen, deuterium, C1-C2 alkyl, C3-C7 straight chain or C3-C7 branched chain alkyl;

[0031] Z is selected from hydrogen, 4-fluoro, 4-hydroxy, 2-(2,6-dichlorophenylamino), 4-isobutyl, 3-fluoro-4-phenyl, 3-phenoxy, 3-benzoyl, 4-(2-methylallylamino) or 4-(2-oxocyclopentylmethyl);

[0032] Y 1 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy;

[0033] Y 2 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy;

[0034] Y 3 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, hydroxy, ethoxy, nitro, amino, methylamino, dimethylamino, acetamido, fluorine, chlorine, bromine or iodine;

[0035] Y 4 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy;

[0036] Y 5 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine or iodine.

[0037] The first aspect of the technical solution of the present invention also provides a class of ibuprofen triazole thiol derivatives selected from the following compounds:

[0038]

[0039]

[0040] The second aspect of the technical solution of the present invention provides a method for preparing an ibuprofen triazole thiol derivative, characterized in that its preparation reaction is as follows:

[0041]

[0042] Wherein, R is selected from: hydrogen, deuterium, C1-C2 alkyl, C3-C7 straight chain or C3-C7 branched chain alkyl;

[0043] R 1 Selected from: hydrogen, C1-C2 alkyl, C3-C7 straight chain or C3-C7 branched chain alkyl;

[0044] Y 1 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy;

[0045] Y 2 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy;

[0046] Y 3 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, hydroxy, ethoxy, nitro, amino, methylamino, dimethylamino, acetamido, fluorine, chlorine, bromine or iodine;

[0047] Y 4 is selected from the group consisting of: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy or ethoxy;

[0048] Y 5 Selected from: hydrogen, deuterium, C1-C2 alkyl, nitro, amino, fluorine, chlorine, bromine, iodine;

[0049] Z is selected from hydrogen, 4-fluoro, 4-hydroxy, 2-(2,6-dichlorophenylamino), 4-isobutyl, 3-fluoro-4-phenyl, 3-phenoxy, 3-benzoyl, 4-(2-methylallylamino) or 4-(2-oxocyclopentylmethyl).

[0050] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition containing the compound described in the first aspect and a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition contains a therapeutically effective amount of the ibuprofen triazole thiol derivative and a pharmaceutically acceptable salt thereof of the present invention, and optionally a pharmaceutical carrier. The pharmaceutical carrier referred to herein refers to a pharmaceutical carrier commonly used in the pharmaceutical field; the pharmaceutical composition can be prepared according to methods well known in the art. The compound of the present invention and its pharmaceutically acceptable salt can be combined with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to form any dosage form suitable for human or animal use. The content of the compound of the present invention and its pharmaceutically acceptable salt in the pharmaceutical composition is generally 0.1% to 95% by weight.

[0051] The compounds of the present invention and their pharmaceutically acceptable salts or pharmaceutical compositions containing the same can be administered in unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0052] The dosage form can be a liquid dosage form, a solid dosage form, or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including O / W, W / O, and multiple emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), and sprays; semisolid dosage forms can be ointments, gels, pastes, and the like.

[0053] The compounds of the present invention and pharmaceutically acceptable salts thereof can be prepared into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle delivery systems.

[0054] In order to prepare the compounds of the present invention and pharmaceutically acceptable salts thereof into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, and glidants. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropyl alcohol, etc.; binders can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, etc.; lubricants and glidants can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0055] The tablets can be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

[0056] To prepare the dosing unit as a capsule, the active ingredient, the compound of the present invention and its pharmaceutically acceptable salt, can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard or soft capsule. Alternatively, the active ingredient, the compound of the present invention and its pharmaceutically acceptable salt, can be first mixed with a diluent, a binder, and a disintegrant to form granules or pellets, which can then be placed in a hard or soft capsule. The diluents, binders, wetting agents, disintegrants, and glidants used to prepare tablets of the compound of the present invention and its pharmaceutically acceptable salt can also be used to prepare capsules of the compound of the present invention and its pharmaceutically acceptable salt.

[0057] To prepare the compounds of the present invention and their pharmaceutically acceptable salts into injections, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of solubilizers, cosolvents, pH adjusters, and osmotic pressure regulators commonly used in the art can be added. Examples of solubilizers or cosolvents include poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, and the like; pH adjusters include phosphates, acetates, hydrochloric acid, sodium hydroxide, and the like; and osmotic pressure regulators include sodium chloride, mannitol, glucose, phosphates, acetates, and the like. For preparations of lyophilized powder injections, mannitol, glucose, and the like can also be added as support agents. Furthermore, colorants, preservatives, fragrances, flavoring agents, or other additives can be added to the pharmaceutical formulations, if desired. To achieve the intended use and enhance the therapeutic effect, the drugs or pharmaceutical compositions of the present invention can be administered by any known method.

[0058] The fourth aspect of the technical solution of the present invention is to provide the use of the ibuprofen triazole thiol derivative (I) or ibuprofen triazole thione derivative (II) and pharmaceutically acceptable salts thereof and the pharmaceutical composition described in the fourth aspect in the preparation of novel coronavirus 3CL protease inhibitors:

[0059]

[0060] Beneficial technical effects:

[0061] The ibuprofen triazole thiol derivative (I) or ibuprofen triazole thione derivative (II) of the present invention is a new class of compounds with novel coronavirus 3CL protease inhibitory activity; they can be used to prepare drugs for treating novel coronavirus. DETAILED DESCRIPTION

[0062] The following examples are intended to illustrate the present invention rather than to further limit the present invention.

[0063] Example 1

[0064] Preparation of (E)-4-(4-hydroxy-3-methoxyphenylmethyleneamino)-5-benzyl-1,2,4-triazole-3-thiol

[0065]

[0066] 2.0 mmol of 4-amino-3-benzyl-1H-1,2,4-triazole-5(4H)-thione and 2.2 mmol of vanillin were refluxed for 5 h, filtered, washed with dichloromethane or ethanol, recrystallized from ethanol, and dried to obtain (E)-4-(4-hydroxy-3-methoxyphenylmethyleneamino)-5-benzyl-1,2,4-triazole-3-thiol as a white solid with a yield of 71.8% and mp of 189-191°C. 1 H NMR (400MHz, DMSO-d6) δ: 3.87 (s, 3H, OCH3), 4.15 (s, 2H, CH2), 6.94 (d, J = 8.0Hz, 1H, C6H3), 7.34-7. 26(m,6H,C6H3,C6H5),7.44(s,1H,C6H32-H),9.66(s,1H,N=CH),10.05(s,1H,OH),13.84(s,1H,NH); 13 CNMR(100MHz,DMSO-d6)δ:164.03,161.86,151.82,150.67,148.61,135.6 9, 129.37, 128.95, 127.32, 124.95, 123.81, 116.03, 110.48, 56.06, 31.21.

[0067] Example 2

[0068] Preparation of (E)-4-(4-hydroxy-3-methoxyphenylmethyleneamino)-5-(p-fluorobenzyl)-1,2,4-triazole-3-thiol

[0069]

[0070] Prepared according to the method of Example 1: 4-amino-3-(p-fluorobenzyl)-1H-1,2,4-triazole-5(4H)-thione and vanillin were refluxed for 6 h to obtain a white solid (E)-4-(4-hydroxy-3-methoxyphenylmethyleneamino)-5-(p-fluorobenzyl)-1,2,4-triazole-3-thiol with a yield of 83.7% and an mp of 210-212°C. 1 H NMR (400MHz, DMSO-d6) δ: 3.85 (s, 3H, OCH3), 4.12 (s, 2H, CH2), 6.91 (d, J = 8.0Hz, 1H, C6H3), 7.13 (t, J = 8.4Hz, 2H, C6H4), 7.27 ( d,J=8.0Hz,1H,C6H3),7.35-7.31(m,2H,C6H4),7.40(s,1H,C6H32-H),9.62(s,1H,N=CH),10.01(s,1H,OH),13.80(s,1H,NH).

[0071] Example 3

[0072] Preparation of (E)-4-(4-hydroxy-3-methoxyphenylmethyleneamino)-5-p-hydroxybenzyl-1,2,4-triazole-3-thiol

[0073]

[0074] Prepared according to the method of Example 1: 4-amino-3-p-hydroxybenzyl-1H-1,2,4-triazole-5(4H)-thione and vanillin were refluxed for 6 h to obtain a pale yellow solid (E)-4-(4-hydroxy-3-methoxyphenylmethyleneamino)-5-p-hydroxybenzyl-1,2,4-triazole-3-thiol with a yield of 84.3% and mp of 213-215°C. 1H NMR (400MHz, DMSO-d6) δ: 3.85 (s, 3H, OCH3), 3.97 (s, 2H, CH2), 6.66 (d, J = 8.0Hz, 2H, C6H4), 6.91 (d, J = 8.0Hz, 1H, C6H32-H), 7.06 (d, J = 8.0 Hz,2H,C6H4),7.27(d,J=8.0Hz,1H,C6H3),7.42(s,1H,C6H32-H),9.30(s,1H,N=CH),9.59(s,1H,OH),10.01(s,1H,OH),13.75(s,1H,NH).

[0075] Example 4

[0076] Preparation of (E)-4-(4-hydroxy-3-ethoxyphenylmethyleneamino)-5-p-hydroxybenzyl-1,2,4-triazole-3-thiol

[0077]

[0078] Prepared according to the method of Example 1: 4-amino-3-p-hydroxybenzyl-1H-1,2,4-triazole-5(4H)-thione and vanillin were refluxed for 6 hours to obtain a white solid (E)-4-(4-hydroxy-3-ethoxyphenylmethyleneamino)-5-p-hydroxybenzyl-1,2,4-triazole-3-thiol with a yield of 70.3% and an mp of 222-224°C. 1 H NMR(400MHz,DMSO-d6)δ:1.39(t,3H,CH3),3.97(s,2H,CH2),4.09(q,2H,OCH2),6.66(d,J=8.0Hz,2H,C6H4),6.92(d,J=8.0Hz,1H,C6H3),7.06( d,J=8.0Hz,2H,C6H4),7.25(d,J=8.0Hz,1H,C6H3),7.40(s,1H,C6H32-H),9.31(s,1H,N=CH),9.58(s,1H,OH),9.93(s,1H,OH),13.75(s,1H,NH).

[0079] Example 5

[0080] Preparation of 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione

[0081]

[0082] 4.6 g (20.0 mmol) of ibuprofen was dissolved in 50 mL of acetonitrile, and 3.2 g (24.0 mmol) of HOBt and 4.6 g (24.0 mmol) of EDCI were added in sequence. After stirring at room temperature for 3 h, 5.1 g (80 mmol) of hydrazine hydrate was added under ice bath. After one and a half hours, the mixture was extracted with ethyl acetate (3 × 20 mL), washed with sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and desolvated to obtain a white solid crude product, which was directly used in the next reaction.

[0083] 2.2 g (10.0 mmol) of the crude product from the previous step and 0.8 g (15.0 mmol) of potassium hydroxide were dissolved in 20 mL of ethanol. A mixture of 1.2 g (15.0 mmol) of carbon disulfide and 4 mL of ethanol was added dropwise under an ice bath. After the addition was complete, the mixture was returned to room temperature and stirred for 2 h. A solid was precipitated, filtered off, and dried. The solid was dissolved in 1.9 g (30.0 mmol) of 80% hydrazine hydrate and refluxed until the reaction was complete. The mixture was cooled and the pH was adjusted to 1 with dilute hydrochloric acid. The precipitated solid was filtered off, washed with water, dried, and recrystallized from methanol to obtain 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione as a white solid, mp 179-181°C, yield 68.7%.

[0084] Example 6

[0085] Preparation of (E)-4-(benzylideneamino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0086]

[0087] Prepared according to the method in the literature [International Journal of Pharmaceutical Education and Research, 2019, 1(02): 47-51]: 0.28 g (1.0 mmol) of 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with 0.11 g (1.0 mmol) of benzaldehyde, and recrystallized to obtain (E)-4-(benzylideneamino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 99~201, yield 82%. 1 H NMR(400MHz,DMSO-d6)δ:1.11-1.14(m,9H,CH3+2×CH3),2.40~2.09(m,1H,CH),2.50(d, 2H, CH2), 3.71 (q, 1H, CH), 7.39~8.00 (m, 9H, Ar-H), 9.84 (s, 1H, N=CH), 13.41 (s, 1H, SH).

[0088] Example 7

[0089] Preparation of (E)-4-((3-nitrobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0090]

[0091] 0.28 g (1.0 mmol) of 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione and 0.17 g (1.1 mmol) of 3-nitrobenzaldehyde were dissolved in 3 mL of acetic acid and refluxed for 4.5 h. The mixture was cooled to precipitate a solid which was filtered to give (E)-4-((3-nitrobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol as a white solid, mp 189-190°C, yield 48.8%; 1 H NMR (400MHz, DMSO-d6) δ: 0.72-0.75 (m, 6H, 2×CH3), 1.59 (d, J=7.1Hz, 3H, CHC H 3), 1.65-1.74 (m, 1H, C H CH2), 2.34 (d, J = 7.1 Hz, 2H, CH2), 4.43 (q, J = 7.1 Hz, 1H, CH), 7.05 (d, J = 7.9 Hz, 2H, benzene ring), 7.16 (d, J = 7.9 Hz, 2H, benzene ring), 7.82-8.55 (m, 4H, benzene ring), 10.14 (s, 1H, NCH), 14.03 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.85, 22.42, 22.49, 29.99, 36.23, 44.59, 122.77, 126.98, 127 .42, 129.63, 131.22, 134.43, 135.00, 139.19, 140.13, 148.71, 154.10, 159.80, 162.14.

[0092] Example 8

[0093] Preparation of (E)-4-((4-nitrobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0094]

[0095] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with 4-nitrobenzaldehyde to obtain white solid (E)-4-((4-nitrobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 196-196°C, yield 80.6%; 1 H NMR (400MHz, DMSO-d6) δ: 0.80-0.70 (m, 6H, 2×CH3), 1.59 (d, J=7.1Hz, 3H, CHC H 3), 1.77-1.66 (m, 1H, C H CH2), 2.35(d, J=7.0Hz, 2H, CHC H 2), 4.45 (q, J=7.1Hz, 1H, C H CH3), 7.06 (d, J = 7.9 Hz, 2H, benzene ring), 7.17 (d, J = 7.9 Hz, 2H, benzene ring), 8.02 (d, J = 8.7 Hz, 2H, benzene ring), 8.32 (d, J = 8.6 Hz, 2H, benzene ring), 10.23 (s, 1H, NCH), 14.05 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.88, 22.46, 22.53, 29.99, 36.09, 44.58, 124.55, 127 .45, 129.63, 129.91, 138.66, 139.11, 140.16, 149.84, 154.25, 159.12, 162.10.

[0096] Example 9

[0097] Preparation of (E)-4-((4-hydroxy-3-methoxybenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0098]

[0099] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with vanillin to obtain a white solid (E)-4-((4-hydroxy-3-methoxybenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 180-182°C, yield 29.2%; 1H NMR (400MHz, DMSO-d6) δ: 0.76-0.79 (m, 6H, 2×CH3), 1.56 (d, J=7.1Hz, 3H, CHC H3 ), 1.69-1.76 (m, 1H, C H CH2), 2.35 (d, J=7.0Hz, 2H, CHC H2 ), 3.83 (s, 3H, OCH3), 4.34 (q, J=7.1Hz, 1H, C H CH3), 6.85-7.30 (m, 7H, benzene ring), 9.44 (s, 1H, OH), 9.98 (s, 1H, NCH), 13.84 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.88, 22.51, 22.57, 30.02, 36.17, 44.61, 55.99, 110.26, 115.91, 123.81, 124.90, 127.46, 129.54, 139.27, 140.05, 148.55, 151.75, 153.80, 161.92, 163.81.

[0100] Example 10

[0101] Preparation of (E)-4-((2-chlorobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0102]

[0103] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with o-chlorobenzaldehyde to obtain a white solid (E)-4-((2-chlorobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 161-162°C, yield 52.7%; 1 H NMR (400MHz, DMSO-d6) δ: 0.74-0.77 (m, 6H, 2×CH3), 1.59 (d, J=7.1Hz, 3H, CHC H3 ), 1.68-1.75 (m, 1H, C H CH2), 2.35(d, J=7.1Hz, 2H, CHC H 2), 4.45 (q, J=7.1Hz, 1H, C HCH3), 7.06 (d, J = 7.9 Hz, 2H, benzene ring), 7.16 (d, J = 7.9 Hz, 2H, benzene ring), 7.44-7.92 (m, 4H, benzene ring), 10.59 (s, 1H, NCH), 14.02 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.94, 22.46, 22.52, 30.02, 36.10, 44.59, 127.41, 127.92, 128 .24, 129.60, 130.34, 130.65, 134.31, 135.45, 139.34, 140.09, 154.32, 156.71, 161.95.

[0104] Example 11

[0105] Preparation of (E)-4-((4-chlorobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0106]

[0107] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with 4-chlorobenzaldehyde to obtain a white solid (E)-4-((4-chlorobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 163-165°C, yield 45.1%; 1 H NMR (400MHz, DMSO-d6) δ: 0.83-0.75 (m, 6H, 2×CH3), 1.60 (d, J=7.1Hz, 3H, CHC H3 ), 1.71-1.77 (m, 1H, C H CH2), 2.37(d, J=7.0Hz, 2H, CHC H2 ), 4.41 (q, J=7.1Hz, 1H, C H CH3), 7.07 (d, J = 7.5 Hz, 2H, benzene ring), 7.16 (d, J = 7.7 Hz, 2H, benzene ring), 7.61 (d, J = 7.6 Hz, 2H, benzene ring), 7.81 (d, J = 7.8 Hz, 2H, benzene ring), 9.85 (s, 1H, NCH), 13.98 (s, 1H, SH); 13C NMR (101MHz, DMSO-d6) δ: 19.86, 22.47, 22.55, 30.00, 36.10, 44.58, 127.44, 129 .58, 129.68, 130.53, 131.52, 137.65, 139.12, 140.11, 153.99, 161.81, 162.01.

[0108] Example 12

[0109] Preparation of (E)-4-((4-bromobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0110]

[0111] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with 4-bromobenzaldehyde to obtain a white solid (E)-4-((4-bromobenzylidene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 140-142°C, yield 53.6%; 1 H NMR (400MHz DMSO-d6) δ: 0.74-0.77 (m, 6H, 2×CH3), 1.57 (d, J=7.1Hz, 3H, CHC H3 ), 1.66-1.75 (m, 1H, C H CH2), 2.35(d, J=7.1Hz, 2H, CHC H 2), 4.39 (q, J=7.1Hz, 1H, C H CH3), 7.04 (d, J = 7.9 Hz, 2H, benzene ring), 7.13 (d, J = 7.9 Hz, 2H, benzene ring), 7.75-7.68 (m, 4H, benzene ring), 9.82 (s, 1H, NCH), 13.95 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.85, 22.48, 22.55, 30.00, 36.10, 44.58, 126.66, 127 .44, 129.58, 130.66, 131.85, 132.62, 139.11, 140.11, 154.00, 161.85, 162.00.

[0112] Example 13

[0113] Preparation of (E)-4-((3-hydroxy-4-methoxybenzylidene)amino)-5-(1-(4-isobutyl-2-phenyl)ethyl)-1,2,4-triazole-3-thiol

[0114]

[0115] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with 3-hydroxy-4-methoxybenzaldehyde to obtain (E)-4-((3-hydroxy-4-methoxybenzylidene)amino)-5-(1-(4-isobutyl-2-phenyl)ethyl)-1,2,4-triazole-3-thiol as a white solid, mp 140-142°C, yield 53.6%; 1 HNMR (400MHz, DMSO-d6) δ: 0.83-0.75 (m, 6H, 2×CH3), 1.57 (d, J=7.0Hz, 3H, CH3), 1.70~1.76 (m, 1H, CH), 2.35 (d, J=6.9Hz, 2H, C H2), 3.84 (s, 3H, OCH3), 4.32 (q, J=7.0Hz, 1H, CH), 7.01~7.28 (m, 7H, benzene ring), 9.40 (s, 1H, OH), 9.47 (s, 1H, NCH), 13.85 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.83, 22.54, 22.59, 30.00, 36.04, 44.62, 56.18, 112.24, 113.65, 123.16, 125.14, 127.49, 129.55, 139.02, 140.12, 147.39, 152.34, 153.79, 161.92, 164.41.

[0116] Example 14

[0117] Preparation of (E)-4-((3-bromo-4-methoxybenzylidene)amino)-5-(1-(4-isobutyl-2-phenyl)ethyl)-1,2,4-triazole-3-thiol

[0118]

[0119] According to the method of Example 7, 4-amino-3-(1-(4-isobutylphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione was reacted with 3-bromo-4-methoxybenzaldehyde to obtain (E)-4-((3-bromo-4-methoxybenzylidene)amino)-5-(1-(4-isobutyl-2-phenyl)ethyl)-1,2,4-triazole-3-thiol as a white solid, mp 153-155°C, yield 27.5%; 1 H NMR (400MHz, DMSO-d6) δ: 0.75-0.78 (m, 6H, 2×CH3), 1.56 (d, J=7.1Hz, 3H, CHC H3 ), 1.69-1.76 (m, 1H, C H CH2), 2.35(d, J=7.1Hz, 2H, CHC H2 ), 3.93 (s, 3H, OCH3), 4.36 (q, J=7.1Hz, 1H, C H CH3), 7.05 (d, J = 7.9 Hz, 2H, benzene ring), 7.12 (d, J = 7.9 Hz, 2H, benzene ring), 7.22-7.94 (m, 3H, benzene ring), 9.61 (s, 1H, NCH), 13.91 (s, 1H, SH); 13 C NMR (101MHz, DMSO-d6) δ: 19.84, 22.50, 22.56, 30.02, 36.18, 44.62, 57.18, 111.89, 113.31, 126.43, 127.42, 129.56, 130.75, 132.63, 139.22, 140.08, 153.86, 159.05, 161.77, 162.01.

[0120] Example 15

[0121] Preparation of (E)-4-((arylmethylene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0122]

[0123] Example 16

[0124] Preparation of (E)-4-((arylmethylene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0125]

[0126] Example 17

[0127] Preparation of (E)-4-((arylmethylene)amino)-5-(1-(4-isobutylphenyl)ethyl)-1,2,4-triazole-3-thiol

[0128]

[0129] Example 18

[0130] Preparation of 4-amino-3-(1-(3-fluoro-4-biphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione

[0131]

[0132] Prepared according to the method of literature [Bioorganic & Medicinal Chemistry, 2016, 24(4): 858-872].

[0133] Example 19

[0134] Preparation of (E)-4-(((4-(dimethylamino)benzylidene)amino)-5-(1-(3-fluoro-4-biphenylyl)ethyl)-1,2,4-triazole-3-thiol

[0135]

[0136] Prepared according to the method of the literature [International Journal of Pharmaceutical Sciences and Research, 2017, 8(4):1598]: 0.31g (1mmol) 4-amino-3-(1-(3-fluoro-4-biphenyl)ethyl)-1H-1,2,4-triazole-5(4H)-thione and 0.15g (1mmol) 4-(dimethylamino)benzaldehyde, reacted for 6h to obtain red solid (E)-4-(((4-(dimethylamino)benzylidene)amino)-5-(1-(3-fluoro-4-biphenyl)ethyl)-1,2,4-triazole-3-thiol, mp 116~118℃, yield 70%, 1 H-NMR (400MHz, DMSO-d6) δ: 1.18 (d, 3H, CH3), 4.55 (q, 1H, CH), 6.77-8.57 (m, 12H, Ar-H), 9.68 (s, 1H, CH=N), 14.10 (s, 1H, SH).

[0137] Example 20

[0138] Preparation of (E)-4-((arylmethylene)amino)-5-(1-(3-fluoro-4-biphenyl)ethyl)-1,2,4-triazole-3-thiol

[0139]

[0140] Example 21

[0141] Anti-novel coronavirus 3CL protease activity of ibuprofen triazole thiol derivatives

[0142] 1 Experimental Principle

[0143] The detection method uses fluorescence resonance energy transfer (FRET). The detection principle is as follows: Edans is the fluorescence donor (Donor), and Dabcyl is the fluorescence acceptor (Acceptor), also known as a quencher. The absorption spectra of these two fluorescent groups overlap to a certain extent. When the distance between the two fluorescent groups is appropriate (generally 7-10nm), the fluorescence energy is transferred from the donor to the acceptor, causing the fluorescence intensity of the donor fluorescent molecule to decrease. Edans and Dabcyl are attached to the two ends of the natural substrate of the 2019-nCoV Mpro / 3CLpro protease, namely Dabcyl-KTSAVLQSGFRKME-Edans. When the 2019-nCoV Mpro / 3CLpro protease does not cleave the substrate, the two groups are close enough to undergo fluorescence resonance energy transfer, meaning that Dabcyl quenches the fluorescence of Edans, resulting in undetectable fluorescence. Once the substrate is cleaved by the 2019-nCoV Mpro / 3CLpro protease, the ends of the peptide separate, and the two groups separate. Edans' fluorescence is no longer quenched by Dabcyl, allowing detection. This allows for highly sensitive fluorescence detection of the 2019-nCoV Mpro / 3CLpro protease activity. If a 2019-nCoV Mpro / 3CLpro inhibitor is added to the reaction system, fluorescence generation is suppressed, and the fluorescence intensity is inversely proportional to the inhibitory effect of the inhibitor, allowing detection of the inhibitory effect of the 2019-nCoV Mpro / 3CLpro protease inhibitor. The maximum excitation wavelength of Edans is 340 nm, and the maximum emission wavelength is 490 nm.

[0144] 2 Experimental methods

[0145] 2.1 Preparation of samples and positive drugs

[0146] Take appropriate amounts of the inhibitor sample to be tested and the positive drug Ebselen, and prepare a solution of appropriate concentration using DMSO.

[0147] 2.2 Assay Reagent Preparation

[0148] Prepare an appropriate amount of Assay Reagent based on the number of samples (including relevant controls). For every 1 μl of 2019-nCoVMpro / 3CLpro, add 92 μl of Assay Buffer to prepare an Assay Reagent sufficient to test one sample.

[0149] 2.3 Sample testing

[0150] Example samples were tested in a 96-well black plate. 93 μl of Assay Reagent and 5 μl of sample were added to each sample well. 93 μl of Assay Reagent and 5 μl of DMSO were added to the model well. 93 μl of Assay Buffer and 5 μl of DMSO were added to the blank control. Mix thoroughly by shaking on a shaker for 1 minute. Quickly add 2 μl of Substrate to each well and shake on a shaker for 1 minute to mix thoroughly. Incubate at 37°C in the dark for 15-20 minutes, then use a multifunctional enzyme label for fluorescence measurement. Excitation wavelength was 340 nm, and emission wavelength was 490 nm.

[0151] 3. Test samples

[0152] Example samples; the positive control drug is ebselen.

[0153] 4 Activity Results

[0154] The 2019-nCoV Mpro / 3CLpro protease inhibitory activity in the reaction system was evaluated at a sample detection concentration of 50 μg / ml. Compounds with an inhibition rate greater than 50% were rescreened three times, and their IC values ​​were calculated based on the rescreening results. 50 The optimal experimental results are shown in Table 1.

[0155]

[0156] Table 1 Inhibitory activity and IC of compounds (50.0 μg / mL) against 2019-nCoV Mpro / 3CLpro protease 50 (μg / mL)

[0157] Y Inhibition rate / % <![CDATA[IC 50 ,μg / mL]]> <![CDATA[3-NO2]]> 72.36±7.99 31.5±8.64 <![CDATA[3-OCH3-4-OH]]> 91.28±2.45 23.84±3.59 2-Cl 97.37±2.07 14.78±2.53 4-Cl 66.28±2.92 34.14±5.04

[0158] Ibuprofen triazole thiol derivatives have anti-novel coronavirus 3CL protease activity and can be used to prepare novel coronavirus 3CL protease inhibitors and drugs for treating novel coronavirus.

Claims

1. Use of a class of ibuprofen triazole thiol derivatives and pharmaceutically acceptable salts thereof in the preparation of anti-coronavirus 3CL protease inhibitors; the ibuprofen triazole thiol derivatives are selected from the following compounds:

2. Use of a class of ibuprofen triazole thiol derivatives and pharmaceutically acceptable salts thereof in the preparation of anti-coronavirus drugs; the ibuprofen triazole thiol derivatives are selected from the following compounds:

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