Dual-mechanism deep fungal infection resisting medicine

Abstract

The invention discloses a dual-mechanism drug for resisting deep fungal infection. The compound is a sulfur-containing triazole compound and a salt thereof, the compound has one of the following structural formulas: # imgabs 0 # imgabs 1 #, in which R is selected from a hydrogen atom or C1-C6 alkyl; a is selected from a C1-C6 alkyl group, a C1-C6 fluoroalkyl group, or a group in which one or more atoms in the C1-C6 alkyl group and the C1-C6 fluoroalkyl group are substituted by isotopes; ar represents a substituted or unsubstituted aryl group or heteroaryl group of a 5-14 membered ring; heteroatoms on the heteroaryl are selected from N, S and O; the substituent group is selected from halogen atoms or C1-C6 alkyl groups. The antifungal drug in the general formula I not only can inhibit fungus 14alpha-sterol demethylase like a traditional conazole drug, but also can inhibit cysteine-containing enzyme of fungi through the contained disulfide bond, so that a dual-action mechanism is exerted.

Description

Technical Field

[0001] The invention belongs to the field of medicine and relates to a class of dual-mechanism anti-deep fungal infection drugs. Background Art

[0002] Fungal infection is a common infectious disease. In recent years, due to the widespread development of organ transplantation surgery, a significant increase in the use of immunosuppressants, radiotherapy and chemotherapy for patients with malignant tumors, and serious infections such as tuberculosis and AIDS, the number of immunocompromised patients has continued to increase. Many fungi that are not pathogenic can cause infection as opportunistic pathogens and even become an important cause of death in immunocompromised patients. According to The Lancet Infectious Diseases (2024, 24, e428-e438), 3.75 million people die worldwide each year due to fungal infections. Unfortunately, the existing types of antifungal drugs are obviously insufficient, and there are only a few commonly used drugs for deep fungal infections in clinical practice, and the problem of drug resistance is becoming increasingly obvious. Therefore, the development of new antifungal drugs is imminent.

[0003] It is reported in the literature that the sulfur atom in the disulfide can undergo sulfur exchange reaction with the enzyme containing cysteine ​​in the structure of fungi and bacteria, thereby inhibiting the activity of the enzyme of fungi and bacteria and affecting the survival of bacteria and fungi. The large amount of glutathione contained in human plasma can neutralize this effect and protect humans from being affected (Biochimica et BiophysicaActa, 1998, 233). Inspired by this mechanism of action, the present invention combines the conazole pharmacophore and the disulfide pharmacophore to design, synthesize and evaluate a new class of antifungal drugs. Summary of the invention

[0004] The purpose of the present invention is to provide a dual-mechanism anti-deep fungal infection drug. This type of antifungal drug can inhibit fungal 14α-sterol demethylase like traditional conazole drugs, and the disulfide bonds contained can inhibit fungal cysteine-containing enzymes, thereby exerting a dual mechanism of action. In addition, by introducing aromatic groups, especially nitrogen-containing heterocycles, into the side chain, it can better combine with the enzyme target 14α-sterol demethylase through multiple interaction forces, thereby obtaining excellent in vitro activity. In vitro experimental results show that this series of compounds are effective against various Candida, Cryptococcus and Aspergillus, and have a broad spectrum of antifungal activity, especially for various drug-resistant bacteria isolated from clinical settings, with outstanding effects. Animal experimental results show that some compounds are more active against Candida and Aspergillus than fluconazole.

[0005] The objective of the present invention is achieved through the following technical solutions:

[0006] In the first aspect, the present invention provides a class of sulfur-containing triazole compounds and salts thereof, wherein the compounds have the following general formula I:

[0007]

[0008] Wherein, A is selected from C1-C6 alkyl, C1-C6 fluoroalkyl, or a group in which one or more atoms of C1-C6 alkyl or C1-C6 fluoroalkyl are substituted by their isotopes;

[0009] Ar represents a substituted or unsubstituted 5-14 membered aryl or heteroaryl group; the heteroatom on the heteroaryl group is selected from N, S, and O; and the substituent is selected from a halogen atom or a C1-C6 alkyl group.

[0010] As one embodiment of the present invention, A is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, difluoroethyl or isotope-substituted ethyl.

[0011] As one embodiment of the present invention, Ar is selected from one of the following groups: phenyl, halogenated phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, isoxazolyl, imidazolyl, thiazolyl, furanyl, pyrazolyl, oxazolyl, pyrrolyl, indolyl, isoindolyl, quinolyl, isoquinolyl.

[0012] In a second aspect, the present invention also provides a class of sulfur-containing triazole compounds and salts thereof, wherein the compound has one of the following structural formulas:

[0013]

[0014] Wherein, R is a hydrogen atom or a C1-C6 alkyl group.

[0015] As one embodiment of the present invention, the salt of the above-mentioned sulfur-containing triazole compound is selected from one of hydrochloride, hydrobromide, hydroiodide, formate, acetate, benzoate, benzenesulfonate, camphorsulfonate, citrate, edisylate, fumarate, glucoheptonate, gluconate, glucuronate, isethionate, lactate, lactobionate, dodecyl sulfate, malate, maleate, methanesulfonate, naphthoate, naphthoate, naphthalenesulfonate, nitrate, stearate, oleate, oxalate, pamoate, phosphate, hydrogen phosphate, dihydrogen phosphate, polygalacturonate, succinate, sulfate, sulfosalicylate, tartrate, p-toluenesulfonate, and trifluoroacetate.

[0016] As some implementation examples of the present invention, the sulfur-containing triazole compound having antifungal activity is preferably selected from the following compounds:

[0017]

[0018] (2R,3R)-2-(2,5-difluorophenyl)-3-mercapto-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0019]

[0020] (2R,3R)-2-(2,5-difluorophenyl)-3-((furan-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0021]

[0022] (2R,3R)-3-((2,4-dichlorobenzyl)disulfanyl)-2-(2,5-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0023]

[0024] (2R,3R)-2-(2,5-difluorophenyl)-3-(phenethyldisulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0025]

[0026] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyridin-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0027]

[0028] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyridin-3-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0029]

[0030] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyridin-4-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0031]

[0032] (2R,3R)-2-(2,5-difluorophenyl)-3-(((3-fluoropyridin-2-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0033]

[0034] (2R,3R)-2-(2,5-difluorophenyl)-3-(((5-fluoropyridin-3-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0035]

[0036] (2R,3R)-2-(2,5-difluorophenyl)-3-(((6-fluoropyridin-2-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0037]

[0038] (2R,3R)-2-(2,5-difluorophenyl)-3-(((2-fluoropyridin-3-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0039]

[0040] methyl

[0041] 6-((((2R,3R)-3-(2,5-difluorophenyl)-3-hydroxy-4-(1H-1,2,4-triazol-1-yl)butan-2-yl)disulfanyl)methyl)picolinate

[0042]

[0043] methyl

[0044] 6-((((2R,3R)-3-(2,5-difluorophenyl)-3-hydroxy-4-(1H-1,2,4-triazol-1-yl)butan-2-yl)disulfanyl)methyl)nicotinate

[0045]

[0046] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyrimidin-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0047]

[0048] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyrazin-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0049]

[0050] (2R,3R)-2-(2,5-difluorophenyl)-3-((thiazol-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0051]

[0052] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-2-yl)ethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0053]

[0054] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-4-yl)ethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0055]

[0056] (2R,3R)-3-((2-(1H-pyrazol-1-yl)ethyl)disulfanyl)-2-(2,5-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0057]

[0058] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyrazin-2-yl)ethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0059]

[0060] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-4-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0061]

[0062] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-methyl-1-(pyridin-4-yl)propan-2-yl)disulfaneyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0063]

[0064] (2R,3R)-2-(2,5-difluorophenyl)-3-((1-(pyridin-4-yl)butan-2-yl)disulfaneyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0065]

[0066] (2R,3R)-2-(2,5-difluorophenyl)-3-((3-(pyridin-2-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0067]

[0068] (2R,3R)-2-(2,5-difluorophenyl)-3-((3-(pyridin-3-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0069]

[0070] (2R,3R)-2-(2,5-difluorophenyl)-3-((3-(pyridin-4-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0071]

[0072] (2R,3R)-2-(2,5-difluorophenyl)-3-((4-(pyridin-4-yl)butyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0073]

[0074] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-4-yl)ethyl-2-d)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0075]

[0076] (2R,3R)-3-((2,2-difluoro-2-phenylethyl)disulfanyl)-2-(2,5-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0077]

[0078] (2R,3R)-2-(2,5-difluorophenyl)-3-(methylthio)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0079]

[0080] (2R,3R)-2-(2,5-difluorophenyl)-3-(methylsulfinyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0081]

[0082] (2R,3R)-2-(2,5-difluorophenyl)-3-(methylsulfonyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0083] In a third aspect, the present invention also provides isotope-labeled forms of the above-mentioned compounds; the isotope-labeled compounds have the structure shown in the above-mentioned structural formula, and one or more atoms in the structural formula are replaced by their isotopes. Any formula or structure given in the present disclosure is also intended to represent the unlabeled form and the isotope-labeled form of the compound. The isotope-labeled compound has the structure described by the formula given herein, except that one or more atoms are replaced by atoms with a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, including but not limited to 2 H (deuterium, D), 3 H(tri), 11 C, 13 C, 14 C, 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl and 125I. Specifically: the structure having the above structure is (as shown in formula I), and one or more hydrogen atoms in the structural formula are 2 H or 3 H, or one or more carbon atoms in the structural formula are replaced 11 C. 13 C or 14 C, or one or more nitrogen atoms in the structural formula are replaced 15 N is substituted, or one or more fluorine atoms in the structural formula are 18 F is substituted, or one or more phosphorus atoms in the structural formula are 31 P or 32 P is substituted, or one or more sulfur atoms in the structural formula are 35 S, or one or more chlorine atoms in the structural formula are replaced 36 Cl is substituted, or one or more iodine atoms in the structural formula are 125 I substitution. Various isotope-labeled compounds of the present disclosure, for example, compounds in which radioactive isotopes such as 3 H, 13 C and 14 Such isotopically labeled compounds can be used for metabolic studies, reaction kinetics studies, detection or imaging techniques.

[0084] In a fourth aspect, the present invention also provides a method for preparing the sulfur-containing triazole compound of the above-mentioned general formula I; the method comprises the following steps: the thiol compound L-1 reacts with a disulfide compound (R1SSR1) to generate an intermediate II, and then reacts with different thiols Ar-A-SH to obtain the disulfide compound.

[0085]

[0086] Among them, R1 in the disulfide compound (R1SSR1) is preferably a self-electron withdrawing group, such as 2,2'-disulfide dipyridine (CAS NO 2127-03-9), 4,4'-disulfide dipyridine (CAS NO 2645-22-9), 3,3'-disulfide dipyridine (CAS NO 24367-50-8), 1,2-di(pyrimidin-2-yl) disulfane (CAS NO 15718-46-4), 4,4'-dinitrodiphenyl disulfide (CAS NO 100-32-3). The solvent is preferably selected from methanol, ethanol, ethyl acetate, tetrahydrofuran, DMSO, dichloromethane, chloroform, acetonitrile, etc.

[0087] In a fifth aspect, the present invention also provides the use of the aforementioned sulfur-containing triazole compounds and pharmaceutically acceptable salts thereof in the preparation of antifungal drugs. The results of in vitro and in vivo antifungal activity experiments show that the compounds of the present invention exhibit good antifungal activity characteristics. Therefore, the compounds of the present invention and their salts can be used to prepare antifungal drugs.

[0088] As an embodiment of the present invention, the antifungal drug includes at least one of an anti-Candida drug, an anti-cryptococcal drug, and an anti-Aspergillus drug.

[0089] In a sixth aspect, the present invention also provides a pharmaceutical composition comprising at least one of the aforementioned compounds or a pharmaceutically acceptable salt thereof, or an isomer thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

[0090] As an embodiment of the present invention, the pharmaceutical composition can be a tablet, dispersible tablet, lozenge, orodisintegrating tablet, sustained-release tablet, capsule, soft capsule, pill, granule, injection, powder injection or aerosol, etc.

[0091] Compared with the prior art, the present invention has the following beneficial effects:

[0092] 1) Through optimized design, the series of compounds obtained by the present invention have excellent antifungal effects, especially against drug-resistant fungi. The results of in vitro tests and animal experiments show that the effects are significantly better than fluconazole. At the same time, the compounds have a wide antibacterial spectrum, covering clinically common Candida, Aspergillus and Cryptococcus.

[0093] 2) The study found that when mice were intraperitoneally injected with a dose of 10 mg / kg of compound L-20, free metabolite L-1 could be detected in plasma at 5 minutes and 60 minutes, but its concentration was very low. After adding dithiothreitol (DTT) for reduction, a large amount of free L-1 could be monitored. This phenomenon shows that L-1 mainly exists in the form of disulfide bonds with endogenous cysteine, and there is very little free L-1. Therefore, it can be reasonably inferred that this series of compounds can also form disulfide bonds with cysteine-containing enzymes in fungi, thereby inhibiting the activity of fungal-related enzymes, and a large amount of glutathione in the human body can neutralize this effect, thereby protecting the human body from the effects of this effect (see reference: The mode of action of allicin: trapping of radicals and interaction with thi olcontaining proteins, Biochimica et Biophysica Acta, 1998, 1379, 233). This type of antifungal drug can also inhibit fungal 14α-sterol demethylase like traditional conazole drugs, thus exerting a dual mechanism of action.

[0094] 3) Inserting methylene between the disulfide groups of L-20, we synthesized compounds M-1 and M-2 (Comparative Examples 1 and 2) with similar structures. These compounds are metabolically stable and have in vitro activities similar to those of L-20. However, they cannot be metabolized into L-1 and cannot exert a dual mechanism of action. Animal experimental results confirmed that the antifungal activity of L-20 is superior to that of M-1 and M-2, with statistical differences. This proves that our design is reasonable and effective, and the disulfide bond is a necessary group for exerting the drug effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0095] Other features, objects and advantages of the present invention will become more apparent from the detailed description of non-limiting embodiments made with reference to the following drawings:

[0096] Figure 1 The results show the effects of compounds L-20, M-1 and M-2 on Candida albicans in an immunosuppressive animal model (cyclophosphamide 100 mg / kg). DETAILED DESCRIPTION

[0097] The present invention is described in detail below in conjunction with embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, some adjustments and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0098] Example 1. Preparation of Compound L-1

[0099] (2R,3R)-2-(2,5-difluorophenyl)-3-mercapto-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0100]

[0101] Dissolve 1-(((2R,3S)-2-(2,5-difluorophenyl)-3-methyloxiran-2-yl)methyl)-1H-1,2,4-triazole (37 g, CAS241479-73-2), 141 g of methyl 3-mercaptopropanoate and 589 mol of sodium methoxide in 600 mL of methanol and heat to reflux for three hours. Then add 35.4 g of methyl 3-mercaptopropanoate and 79.6 mol of sodium methoxide, and reflux for one hour. After cooling, add 880 mL of 1M hydrochloric acid. Then extract with 2.5 L of DCM, wash with brine, dry with magnesium sulfate, spin dry the solvent, and recrystallize in ethyl acetate to obtain compound L-1.

[0102] 1 H NMR (400MHz, CDCl3) δ7.88(s,1H),7.83(s,1H),7.17-7.08(m,1H),6.99-6.92(m,1H),6.92-6.85(m ,1H),5.05-4.83(m,2H),4.12(q,J=7.1Hz,1H),1.96(dd,J=10.3,1.0Hz,1H),1.18(d,J=6.9Hz,3H).

[0103] Example 2: Preparation of Intermediate II

[0104] (2R,3R)-2-(2,4-difluorophenyl)-3-(pyridin-2-yldisulfaneyl)-1-(1H-1,2,4-triazol-1-yl)buta n-2-ol

[0105]

[0106] Compound L-1 (2.85 g, 10 mmol, 1.0 equiv.) and 2,2'-disulfide dipyridine (4.40 g, 20 mmol, 2.0 equiv.) were dissolved in methanol (20 mL) and stirred at room temperature for 12 hours. After the reaction was completed, the solvent was removed by vortexing, and petroleum ether and ethyl acetate solution (50 mL, petroleum ether: ethyl acetate = 5:1) were added. After stirring, a yellow solid was generated. After filtering, the filtrate was dried by vortexing, and intermediate II (2.30 g, yield: 81%) was obtained after column chromatography purification. 1H NMR (400MHz, CDCl3) δ8.73-8.66(m,1H),7.93(s,1H),7.76(s,1H),7.66(td,J=7.7,1.8Hz,1H),7.49(d,J=8.0Hz,1H),7.31-7.23(m,1H) ,7.24-7.17(m,1H),7.03-6.95(m,1H),6.94-6.86(m,1H),5.14(dd,J=168.0,14.0Hz,2H),3.80(q,J=6.9Hz,1H),1.25(d,J=6.4Hz,3H).

[0107] Example 3: Preparation of Compound L-2

[0108]

[0109] (2R,3R)--2-(2,5-difluorophenyl)-3-((furan-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0110] Intermediate II (200 mg, 0.508 mmol, 1.0 equiv.) and furan-2-ylmethanethiol (3.0 equiv.) were dissolved in methanol (10 mL) and stirred at room temperature for 12 hours. After the reaction was completed, the solvent was removed by vortexing and the target product L-2 was obtained as a colorless oily compound (111 mg) by column chromatography.

[0111] 1 H NMR (400MHz, CDCl3) δ7.81 (s, 1H), 7.77 (s, 1H), 7.44 (d, J = 1.8Hz, 1H), 7.08-7.01 (m, 1H), 6.96-6.89 (m, 1H), 6.89- 6.82(m,1H),6.37-6.31(m,2H),4.99-4.88(m,2H),4.07-3.95(m,2H),3.15(q,J=7.1Hz,1H),1.08(d,J=7.1Hz,3H).

[0112] 13C NMR (101MHz, CDCl3) δ158.74 (d, J = 243.3Hz), 153.85 (d, J = 239.7Hz), 151.8 5,150.33,143.81,142.65,130.10(dd,J=15.5,6.8Hz),116.69-116.43(m) ,116.57(dd,J=62.8,8.7Hz),116.03(dd,J=26.3,4.7Hz),110.68,109.23, 78.53(d,J=5.3Hz),56.10(d,J=6.1Hz),54.43(d,J=3.4Hz),36.43,16.89.

[0113] Example 4: Preparation of Compound L-3

[0114] (2R,3R)-3-((2,4-dichlorobenzyl)disulfanyl)-2-(2,5-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0115]

[0116] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and (2,4-dichlorophenyl)methanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-3 (132 mg) was obtained by column chromatography.

[0117] 1 H NMR (400MHz, CDCl3) δ7.80(s,1H),7.78(s,1H),7.45(d,J=2.1Hz,1H),7.35(d,J=8.2Hz,1H),7.28-7.22(m,1H),7.07-7.01( m,1H),6.96-6.89(m,1H),6.89-6.82(m,1H),4.93-4.80(m,2H),4.15-4.03(m,2H),3.22-3.12(m,1H),1.08(d,J=7.1Hz,3H).

[0118] 13C NMR (101MHz, CDCl3) δ158.78 (d, J = 243.6Hz), 153.80 (d, J = 238.9Hz), 151.96, 143.83,135.05,134.24,133.51,132.30,130.11-129.81(m),129.72,127.11 ,116.65(dd,J=59.4,8.8Hz),116.64(t,J=8.5Hz),116.02(dd,J=26.3,4.6Hz ),78.54(d,J=5.3Hz),56.02(d,J=6.3Hz),54.36(d,J=3.8Hz),41.04,16.79.

[0119] Example 5. Preparation of Compound L-4

[0120] (2R,3R)-2-(2,5-difluorophenyl)-3-(phenethyldisulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0121]

[0122] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-phenylethane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-4 (125 mg) was obtained by column chromatography.

[0123] 1 H NMR (400MHz, CDCl3) δ7.81 (d, J = 16.0Hz, 2H), 7.36-7.29 (m, 2H), 7.28-7.21 (m, 3H), 7.12-7.05 (m, 1H), 6.98-6 .91(m,1H),6.91-6.83(m,1H),5.08-4.95(m,2H),3.53-3.43(m,1H),3.14-2.96(m,4H),1.22(d,J=7.3Hz,3H).

[0124] 13C NMR (101MHz, CDCl3) δ158.81 (d, J = 243.4Hz), 153.88 (d, J = 236.2Hz), 151. 88,143.85,139.74,130.32-129.91(m),128.59,126.53,116.84(dd,J=27. 2,8.8Hz),116.65-116.29(m),116.11(dd,J=26.3,4.8Hz),78.64(d,J=5. 2Hz),56.26(d,J=6.3Hz),54.88(d,J=3.7Hz),50.69,40.90,35.51,17.23.

[0125] Example 6: Preparation of Compound L-5

[0126] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyridin-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0127]

[0128] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and pyridin-2-ylmethanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-5 (113 mg) was obtained by column chromatography.

[0129] 1 H NMR (400MHz, CDCl3) δ8.88-8.84 (m, 1H), 8.19 (td, J = 7.8, 1.7Hz, 1H), 8.03 (s, 1H), 7. 84(s,1H),7.81-7.78(m,1H),7.66(ddd,J=7.6,5.5,1.2Hz,1H),7.08(ddd,J=9.3,6.1 ,3.2Hz,1H),6.97(ddd,J=10.8,9.0,4.4Hz,1H),6.90(ddt,J=8.9,7.0,3.5Hz,1H),5. 05-4.84(m,2H),4.33(q,J=13.9Hz,2H),3.52(q,J=7.0Hz,1H),1.18(d,J=7.0Hz,3H).

[0130] 13C NMR (101MHz, CDCl3) δ158.81 (d, J = 244.7Hz), 155.21, 152.83, 150.58, 144.12, 143.73, 142.39, 129.46, 126.34, 124.31, 116.7 8(dd,J=55.4,8.7Hz),116.77(dd,J=8.8,4.2Hz),116.05(dd,J=26.3,4.6Hz),78.00(d,J=5.1Hz),56.40,53.06,40.28,16.36.

[0131] Example 7: Preparation of Compound L-6

[0132] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyridin-3-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0133]

[0134] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and pyridin-3-ylmethanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-6 (128 mg) was obtained by column chromatography.

[0135] 1 H NMR(400MHz, CDCl3)δ8.91(d,J=2.1Hz,1H),8.73(dd,J=5.5,1.5Hz,1H),8.33-8.26(m,1H),8.01(s,1H),7.85-7.77(m,2H),7.09- 7.02(m,1H),6.99-6.92(m,1H),6.92-6.84(m,1H),4.99-4.86(m,2H),4.16-4.03(m,2H),3.36-3.26(m,1H),1.16(d,J=7.0Hz,3H).

[0136] 13C NMR (101MHz, CDCl3) δ158.79 (d, J = 244.6Hz), 153.89 (d, J = 238.8Hz), 151.15, 143.99, 143.65, 142.08, 129.54 (dd, J = 15.5, 6.7Hz), 126.11, 116 .85(dd,J=54.4,9.0Hz),116.84(dd,J=8.3,2.5Hz),115.93(dd,J=26.4,4.7Hz),78.18(d,J=5.3Hz),56.02(d,J=6.2Hz),53.52,39.33,16.71.

[0137] Example 8. Preparation of Compound L-7

[0138] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyridin-4-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0139]

[0140] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and pyridin-4-ylmethanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-7 (122 mg) was obtained by column chromatography.

[0141] 1 H NMR (400MHz, CDCl3) δ8.82 (d, J = 6.1Hz, 2H), 8.03-7.75 (m, 4H), 7.08-7.02 (m, 1H), 7.01-6.93 (m, 1H ),6.93-6.85(m,1H),4.89(s,2H),4.20-4.05(m,2H),3.37(q,J=7.0Hz,1H),1.18(d,J=7.0Hz,3H).

[0142] 13C NMR (101MHz, CDCl3) δ158.80 (d, J = 243.9Hz), 156.22, 153.85 (d, J = 237.6Hz), 151.47, 143.29-142.38 (m), 129.47 (dd, J = 15.4, 6.9Hz), 126.86,116.99-116.82(m),116.93(dd,J=54.0,8.9Hz),115.91(dd,J=26.3,4.5Hz),78.21(d,J=5.1Hz),55.99,53.54,42.09,16.73.

[0143] Example 9: Preparation of Compound L-8

[0144] (2R,3R)-2-(2,5-difluorophenyl)-3-(((3-fluoropyridin-2-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0145]

[0146] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and (3-fluoropyridin-2-yl)methanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-8 was obtained by column chromatography as a colorless viscous liquid with a yield of 38%.

[0147] 1 H NMR (400MHz, CDCl3) δ8.68-8.62(m,1H),7.86(s,1H),7.84(s,1H),7.52(t,J=8.8Hz,1H),7.41-7.32(m,1H),7.23-7.13(m,1 H),7.04-6.95(m,1H),6.95-6.87(m,1H),5.06-4.84(m,2H),4.20-4.07(m,2H),3.55(q,J=7.0Hz,1H),1.15(d,J=7.0Hz,3H).

[0148] 13C NMR (101MHz, CDCl3) δ158.89 (d, J = 242.4Hz), 157.59 (d, J = 258.9Hz), 155.28, 15 2.93,150.81,145.43(d,J=15.6Hz),144.30(d,J=5.7Hz),143.45,130.27-129. 84(m),124.34(d,J=4.2Hz),124.00(d,J=18.7Hz),116.93-116.61(m),116.60- 116.42(m),116.36-116.07(m),77.81,56.59(d,J=5.6Hz),51.78,36.52,15.59.

[0149] Example 10: Preparation of Compound L-9

[0150] (2R,3R)-2-(2,5-difluorophenyl)-3-(((5-fluoropyridin-3-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0151]

[0152] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and (5-fluoropyridin-3-yl)methanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-9 was obtained by column chromatography as a colorless viscous liquid with a yield of 44%.

[0153] 1 H NMR (400MHz, CDCl3) δ8.73-8.67(m,1H),7.99(s,1H),7.78(s,1H),7.67(td,J=7.7,1.7Hz,1H),7.49(d,J=8.0Hz,1H),7.23-7. 16(m,1H),7.04-6.95(m,1H),6.95-6.87(m,1H),5.15(dd,J=166.5,14.2Hz,2H),3.81(q,J=6.9Hz,1H),1.25(d,J=6.9Hz,3H).

[0154] 13C NMR (101MHz, CDCl3) δ159.23, 158.83 (d, J = 243.5Hz), 154.51 (d, J = 241.5Hz), 150.64, 149.52, 143.66, 137.37, 130.32-129.96 (m ),122.89,122.15,116.94-116.60(m),116.62-116.35(m),116.35-115.91(m),76.86,56.94(d,J=5.7Hz),53.41,51.06,15.73.

[0155] Example 11. Preparation of Compound L-10

[0156] (2R,3R)-2-(2,5-difluorophenyl)-3-(((6-fluoropyridin-2-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0157]

[0158] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and (6-fluoropyridin-2-yl)methanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-10 was obtained by column chromatography as a colorless viscous liquid with a yield of 47%.

[0159] 1 H NMR (400MHz, CDCl3) δ8.02 (s, 1H), 7.91-7.76 (m, 2H), 7.29 (d, J = 2.2Hz, 1H), 7.12-7.04 (m, 1H), 6.98-6.91 (m, 1H), 6. 91-6.82(m,1H),5.31(d,J=0.8Hz,1H),4.98(s,2H),4.09-3.96(m,2H),3.37(q,J=7.1Hz,1H),1.11(d,J=7.1Hz,3H).

[0160] 13C NMR (101MHz, CDCl3) δ162.07(d,J=421.5Hz),161.77,156.52,156.37(d,J=239.6Hz),152.79,151.21,143.77,141.95(d,J= 8.2Hz),121.01,116.75(dd,J=27.3,9.0Hz),116.56-116.19(m),116.19-115.79(m),108.41,108.05,78.29,44.03,16.51.

[0161] Example 12: Preparation of Compound L-11

[0162] (2R,3R)-2-(2,5-difluorophenyl)-3-(((2-fluoropyridin-3-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0163]

[0164] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and (2-fluoropyridin-3-yl)methanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-11 was obtained by column chromatography as a colorless viscous liquid with a yield of 31%.

[0165] 1 H NMR (400MHz, CDCl3) δ8.19 (d, J = 4.7Hz, 1H), 7.93 (s, 1H), 7.89-7.79 (m, 2H), 7.25-7.20 (m, 1H), 7.08-7.00 (m, 1H) ),6.99-6.91(m,1H),6.91-6.83(m,1H),4.94(s,2H),4.01(s,2H),3.26(q,J=7.1Hz,1H),1.11(d,J=7.1Hz,3H).

[0166] 13C NMR (101MHz, CDCl3) δ160.53,151.16,146.89,146.74,143.64,141.77,121.66,120.02,119.72,117 .15-116.81(m),116.81-116.47(m),116.15-115.76(m),78.46,56.21,54.12,53.41,36.27,16.77.

[0167] Example 13: Preparation of Compound L-12

[0168] methyl6-((((2R,3R)-3-(2,5-difluorophenyl)-3-hydroxy-4-(1H-1,2,4-triazol-1-yl)butan-2-yl)disulfanyl)methyl)picolinate

[0169]

[0170] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and methyl 6-(mercaptomethyl)picolinate (3.0 equiv.) were reacted according to Example 3, and the target product L-12 was obtained by column chromatography as a colorless viscous liquid with a yield of 43%.

[0171] 1 H NMR (400MHz, CDCl3) δ8.17(s,1H),8.07(dd,J=7.8,1.1Hz,1H),7.89(t,J=7.8Hz,1H),7.81(s,1H),7.62(dd,J=7.8,1.1Hz,1H),7.14-7.07(m ,1H),6.95-6.89(m,1H),6.89-6.83(m,1H),5.08-4.98(m,2H),4.20(q ,J=13.4Hz,2H),3.92(s,3H),3.41-3.31(m,1H),1.13(d,J=7.0Hz,3H).

[0172] 13C NMR (101MHz, CDCl3) δ165.42, 158.76 (d, J = 243.0Hz), 158.40, 153.97 (d, J = 239.9Hz), 150.52, 147.27, 143.86, 138.00, 129.70, 127.19, 12 3.96,116.73(dd,J=26.3,7.6Hz),116.56-116.21(m),116.00(dd,J=26.5,4.8Hz),78.28(d,J=5.0Hz),56.81,56.75,52.90,44.38,16.65.

[0173] Example 14: Preparation of Compound L-13

[0174] methyl6-((((2R,3R)-3-(2,5-difluorophenyl)-3-hydroxy-4-(1H-1,2,4-triazol-1-yl)butan-2-yl)disulfanyl)methyl)nicotinate

[0175]

[0176] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and methyl 6-(mercaptomethyl)nicotinate (3.0 equiv.) were reacted according to Example 3, and the target product L-13 was obtained by column chromatography as a colorless viscous liquid with a yield of 59%.

[0177] 1 H NMR (400MHz, CDCl3) δ9.37(s,1H),8.37(dd,J=8.1,2.1Hz,1H),7.85(s,1H),7.80(s,1H),7.51(d,J=8.1Hz,1H),7.20-7.10(m,1H),7 .01-6.94(m,1H),6.93-6.86(m,1H),5.02-4.83(m,2H),4.24-4.00(m,2H),3.97(s,3H),3.48(q,J=6.9Hz,1H),1.14(d,J=7.0Hz,3H).

[0178] 13C NMR (101MHz, CDCl3) δ165.33, 161.24, 158.88 (d, J = 243.0Hz), 154.04 (d, J = 239.2Hz), 151.30, 149.92, 143.59, 138.57, 125.06,123.64,116.95-116.59(m),116.59-116.38(m),116.38-116.09(m),77.98,56.46,52.53,52.26,43.34,15.97.

[0179] Example 15. Preparation of Compound L-14

[0180] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyrimidin-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0181]

[0182] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and pyrimidin-2-ylmethanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-14 (115 mg) was obtained by column chromatography.

[0183] 1 H NMR (400MHz, CDCl3) δ8.94(d,J=5.0Hz,2H),7.88(s,1H),7.80(s,1H),7.34(t,J=5.0Hz,1H),7.20-7.14(m,1H),7. 05-6.97(m,1H),6.97-6.90(m,1H),5.09-4.85(m,2H),4.27-4.09(m,2H),3.64-3.55(m,1H),1.17(d,J=7.0Hz,3H).

[0184] 13 C NMR (101MHz, CDCl3) δ 166.49, 158.91 (d, J = 242.3Hz), 157.44, 152.93, 150.58, 143.38, 130. 16-129.73(m),119.65,117.02-116.19(m),77.77,56.62(d,J=5.7Hz),51.71,44.35,15.58.

[0185] Example 16. Preparation of Compound L-15

[0186] (2R,3R)-2-(2,5-difluorophenyl)-3-((pyrazin-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0187]

[0188] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and pyrazin-2-ylmethanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-15 (92 mg) was obtained by column chromatography.

[0189] 1 H NMR (400MHz, CDCl3) δ8.73(dd,J=2.6,1.5Hz,1H),8.70(d,J=1.5Hz,1H),8.58(d,J=2.6Hz,1H),7.84(s,1H),7.81(s,1H),7.15-7.0 7(m,1H),7.02-6.94(m,1H),6.94-6.87(m,1H),5.03-4.85(m,2H),4.18-4.04(m,2H),3.39(q,J=7.0Hz,1H),1.13(d,J=7.0Hz,3H).

[0190] 13 C NMR (101MHz, CDCl3) δ158.86 (d, J = 243.7Hz), 153.95 (d, J = 236.1Hz), 152.99, 151.24, 145.22, 143.72, 143.61, 143.49, 129.92 (d, J = 15. 3Hz),117.09-116.28(m),116.67(dd,J=8.8,3.5Hz),116.35-116.05(m),78.08(d,J=5.2Hz),56.31(d,J=6.0Hz),52.76,41.29,16.13.

[0191] Example 17: Preparation of Compound L-16

[0192] (2R,3R)-2-(2,5-difluorophenyl)-3-((thiazol-2-ylmethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0193]

[0194] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and thiazol-2-ylmethanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-16 (108 mg) was obtained by column chromatography.

[0195] 1 H NMR (400MHz, CDCl3) δ7.84(s,1H),7.82(d,J=3.3Hz,1H),7.78(s,1H),7.38(d,J=3.3Hz,1H),7.15-7.09(m,1H),6.98 -6.92(m,1H),6.91-6.84(m,1H),5.02-4.85(m,2H),4.41-4.24(m,2H),3.45(q,J=7.1Hz,0H),1.15(d,J=7.0Hz,3H).

[0196] Example 18: Preparation of Compound L-17

[0197] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-2-yl)ethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0198]

[0199] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-(pyridin-2-yl)ethane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-17 (122 mg) was obtained by column chromatography.

[0200] 1H NMR (400MHz, CDCl3) δ8.90-8.83(m,1H),8.23(td,J=7.8,1.6Hz,1H),7.98(s,1H),7.79(s,1H),7.74-7.68(m,2H),7.09-7.03(m ,1H),7.00-6.92(m,1H),6.91-6.84(m,1H),4.98(q,J=14.2Hz,2H),3.64-3.41(m,3H),3.35-3.19(m,2H),1.20(d,J=7.0Hz,3H).

[0201] 13 C NMR (101MHz, CDCl3) δ158.73 (d, J = 244.5Hz), 155.93, 155.23-152.70 (m), 15 1.27,143.97,143.45,129.76(dd,J=15.4,7.0Hz),127.04,124.26,116.72( dd,J=65.9,8.8Hz),116.71(dd,J=14.8,8.9Hz),115.95(dd,J=26.3,4.6Hz) ,78.24(d,J=5.2Hz),56.22(d,J=6.0Hz),54.40(d,J=3.6Hz),33.55,16.94.

[0202] Example 19: Preparation of Compound L-18

[0203] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-4-yl)ethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0204]

[0205] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-(pyridin-4-yl)ethane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-18 (92 mg) was obtained by column chromatography.

[0206] 1H NMR (400MHz, CDCl3) δ8.54(d,J=5.0Hz,2H),7.82(s,1H),7.79(s,1H),7.21-7.15(m,2H),7.12-7.06(m,1H),6.99 -6.92(m,1H),6.91-6.83(m,1H),5.06-4.93(m,2H),3.53-3.43(m,1H),3.16-2.96(m,4H),1.22(d,J=7.1Hz,3H).

[0207] Example 20, Preparation of Compound L-19

[0208] (2R,3R)-3-((2-(1H-pyrazol-1-yl)ethyl)disulfanyl)-2-(2,5-difluorophenyl)-1-(1H-1,2,4-tri azol-1-yl)butan-2-ol

[0209]

[0210] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-(1H-pyrazol-1-yl)ethane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-19 (124 mg) was obtained by column chromatography.

[0211] 1 H NMR (400MHz, CDCl3) δ7.87(s,1H),7.81(s,1H),7.56(dd,J=1.9,0.7Hz,1H),7.47(dd,J=2.3,0.7Hz,1H),7.12-7.06(m,1H),6.99 -6.92(m,1H),6.91-6.85(m,1H),4.99(s,2H),4.51(t,J=6.7Hz,2H),3.51-3.44(m,1H),3.31-3.18(m,2H),1.21(d,J=7.1Hz,3H).

[0212] 13C NMR (101MHz, CDCl3) δ158.83 (d, J = 243.4Hz), 153.84 (d, J = 238.8Hz), 151.77, 143.84, 139.88, 130.03, 116.74 (dd, J = 56.1, 8.8Hz), 116.73 (dd,J=8.8,5.0Hz),116.11(dd,J=26.4,4.6Hz),105.63,78.48(d,J=5.2Hz),56.19(d,J=6.3Hz),54.58(d,J=3.3Hz),50.68,38.80,17.07.

[0213] Example 21. Preparation of Compound L-20

[0214] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyrazin-2-yl)ethyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0215]

[0216] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-(pyrazin-2-yl)ethane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-20 (98 mg) was obtained by column chromatography.

[0217] 1 H NMR (400MHz, CDCl3) δ8.49-8.45(m,2H),8.39(d,J=2.2Hz,1H),7.80(s,1H),7.71(s,1H),7.07-7.00(m,1H),6.93- 6.85(m,1H),6.85-6.78(m,1H),5.00-4.91(m,2H),3.44(q,J=7.1Hz,1H),3.26-3.10(m,4H),1.15(d,J=7.1Hz,3H).

[0218] Example 22: Preparation of Compound L-21

[0219] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-4-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazo l-1-yl)butan-2-ol

[0220]

[0221] MePPh3Br (5.35 g, 15 mmol, 1.5 equiv.) was dissolved in THF (40 mL) and added at 0°C under nitrogen protection. t BuOK (1.68 g, 15 mmol, 1.5 equiv.), the system was kept at 0°C for 1 hour. 4-Acetylpyridine (1.21 g, 10 mmol, 1.0 equiv.) was dissolved in THF (10 mL) and added dropwise to the system. After stirring at room temperature for 4 hours, saturated ammonium chloride was added to quench the reaction, and ethyl acetate (3*40 mL) was extracted. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by rotary evaporation. Column chromatography was used for purification to obtain colorless liquid 4-(prop-1-en-2-yl)pyridine (680 mg, yield: 57%).

[0222] 4-(prop-1-en-2-yl)pyridine (680 mg, 5.71 mmol, 1.0 equiv.) was dissolved in DCM (20 mL), and AcSH (0.59 mL, 8.56 mmol, 1.5 equiv.) and I2 (290 mg, 1.14 mmol, 0.2 equiv.) were added to the system. After stirring at room temperature overnight, the mixture was dried by spin drying and purified by column chromatography to obtain a colorless liquid (620 mg, yield: 56%).

[0223] The above colorless liquid (620 mg, 3.18 mmol, 1.0 equiv.) was dissolved in MeOH (20 mL). Under nitrogen protection, potassium carbonate (878 mg, 6.36 mmol, 2.0 equiv.) was added. After stirring at room temperature for 2 hours, water (20 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (3*40 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried. The filtrate was purified by column chromatography to obtain a colorless liquid 2-(pyridin-4-yl)propane-1-thiol (389 mg, yield: 80%). 1 H NMR (400MHz, CDCl3): δ8.54-8.44(m,2H),7.12-7.02(m,2H),2.85(h,J=6.9H z,1H),2.77-2.60(m,2H),1.30(d,J=6.9Hz,4H),1.19(dd,J=8.9,7.7Hz,1H).

[0224]

[0225] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-(pyridin-4-yl)propane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-21 (92 mg) was obtained by column chromatography.

[0226] 1 H NMR (400MHz, CDCl3) δ8.84-8.77(m,2H),7.96(d,J=7.8Hz,1H),7.80(d,J=7 .2Hz,1H),7.74(s,1H),7.72(s,1H),7.11-7.02(m,1H),7.01-6.93(m,1H), 6.92-6.85(m,1H),5.01(s,1H),5.07-4.92(m,1H),3.52-3.42(m,1H),3.39 -3.23(m,2H),3.17-3.03(m,1H),1.47-1.34(m,3H),1.20(d,J=7.1Hz,3H).

[0227] 13 C NMR(101MHz, CDCl3)δ158.77(d,J=244.0Hz),158.00,153.90(d,J=237.5Hz),151.52 ,151.38,143.96,142.18,129.73(d,J=15.2Hz),127.34,127.30,116.98(dd,J=27.1 ,8.6Hz),116.72-116.41(m),116.23-115.72(m),78.44(d,J=5.2Hz),78.36(d,J=4. 9Hz),56.15,55.87,55.83,55.32,46.31,42.10,41.85,20.54,19.80,17.05,16.90.

[0228] Example 23. Preparation of Compound L-22

[0229]

[0230] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 2-methyl-1-(pyridin-4-yl)propane-2-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-22 (114 mg) was obtained by column chromatography.

[0231] 1H NMR (400MHz, CDCl3) δ8.84-8.77(m,2H),8.03(s,1H),7.80(s,1H),7.74-7.68(m,2H),7.12-7.02(m,1H),7.00-6.93(m,1H),6. 92-6.85(m,1H),5.13-5.00(m,2H),3.53(q,J=7.2Hz,1H),3.24-3.08(m,2H),1.44(s,3H),1.39(s,3H),1.22(d,J=7.2Hz,3H).

[0232] 13 C NMR (101MHz, CDCl3) δ158.75 (d, J = 243.8Hz), 156.65, 153.94 (d, J = 239.3Hz), 151.1 8,143.89,141.48,129.72(dd,J=15.2,6.9Hz),128.66,116.96(dd,J=26.8,8.6Hz) ,116.57(dd,J=24.4,9.1Hz),115.92(dd,J=26.4,4.5Hz),78.54(d,J=5.2Hz),57.2 3(d,J=3.8Hz),56.28(d,J=6.1Hz),51.07(d,J=1.7Hz),46.71,27.98,27.02,17.01.

[0233] Example 24: Preparation of Compound L-23

[0234] (2R,3R)-2-(2,5-difluorophenyl)-3-((1-(pyridin-4-yl)pentan-3-yl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0235]

[0236] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 1-(pyridin-4-yl)pentane-3-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-23 (116 mg) was obtained by column chromatography.

[0237] 1H NMR (400MHz, CDCl3) δ8.81-8.75 (m, 2H), 7.96 (d, J = 4.6Hz, 1H), 7.79 (s, 1H ),7.76-7.70(m,2H),7.11-7.02(m,1H),7.01-6.92(m,1H),6.92-6.84(m,1 H),5.04(s,2H),3.53-3.42(m,1H),3.19-2.93(m,2H),2.82-2.66(m,1H),2 .17-2.02(m,2H),1.19(dd,J=7.1,3.1Hz,3H),1.09(td,J=7.3,1.6Hz,3H).

[0238] 13 C NMR (101MHz, CDCl3) δ161.23, 158.77 (d, J = 244.0Hz), 153.91 (d, J = 235.3Hz), 151.4 7,143.93,142.06,129.99-129.70(m),126.48,117.13-116.73(m),116.73-116.33 (m),116.16-115.74(m),78.61-78.39(m),56.24,55.73-55.39(m),53.58(d,J=15. 2Hz),33.61,33.28,33.09,32.82,27.26,26.82,17.18,16.97,11.41(d,J=2.3Hz).

[0239] Example 25. Preparation of Compound L-24

[0240] (2R,3R)-2-(2,5-difluorophenyl)-3-((3-(pyridin-2-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazo l-1-yl)butan-2-ol

[0241]

[0242] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 3-(pyridin-2-yl)propane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-24 (128 mg) was obtained by column chromatography.

[0243] 1H NMR (400MHz, CDCl3) δ8.88-8.82(m,1H),8.32-8.26(m,1H),7.82(s,1H),7.77-7.71(m,2H),7.09-7.03(m,1H),7.00-6.93(m,1H), 6.92-6.85(m,1H),5.01(s,2H),3.53-3.45(m,1H),3.35-3.23(m,2H),2.94-2.78(m,2H),2.37-2.23(m,2H),1.18(d,J=7.1Hz,3H).

[0244] 13 C NMR (101MHz, CDCl3) δ159.97,157.49,153.98(d,J=237.5Hz),150.69,144.46,143.80,142.50,129.91-129.58(m),126.40,124.22,116.73(dd,J= 64.7,8.9Hz),116.72(dd,J=13.7,8.8Hz),115.94(dd,J=26.3,4.6Hz),78 .27(d,J=5.1Hz),56.44(d,J=6.2Hz),54.48,37.73,32.25,28.43,17.04.

[0245] Example 26. Preparation of Compound L-25

[0246] (2R,3R)-2-(2,5-difluorophenyl)-3-((3-(pyridin-3-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazo l-1-yl)butan-2-ol

[0247]

[0248] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 3-(pyridin-3-yl)propane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-25 (92 mg) was obtained by column chromatography.

[0249] 1H NMR (400MHz, CDCl3) δ8.81 (s, 1H), 8.70 (d, J = 5.5Hz, 1H), 8.20 (d, J = 8.0Hz, 1H),7.99(s,1H),7.82(s,1H),7.81-7.78(m,1H),7.10-7.04(m,1H),7.00-6 .93(m,1H),6.92-6.86(m,1H),5.05-4.96(m,2H),3.48(q,J=7.1Hz,1H),3.0 6-2.94(m,2H),2.93-2.76(m,2H),2.23-2.11(m,2H),1.21(d,J=7.1Hz,3H).

[0250] 13 C NMR (101MHz, CDCl3) δ153.92 (d, J = 238.4Hz), 151.09, 144.00, 143.78, 142.33, 141.13, 14 0.27,126.23,78.41(d,J=5.0Hz),56.36(d,J=6.1Hz),54.61,37.98,31.03,29.22,17.10.

[0251] Example 27, Preparation of Compound L-26

[0252] (2R,3R)-2-(2,5-difluorophenyl)-3-((3-(pyridin-4-yl)propyl)disulfanyl)-1-(1H-1,2,4-triazo l-1-yl)butan-2-ol

[0253]

[0254] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and 3-(pyridin-4-yl)propane-1-thiol (3.0 equiv.) were reacted according to Example 3, and the target product L-26 (92 mg) was obtained by column chromatography.

[0255] 1H NMR (400MHz, CDCl3) δ8.79(d,J=6.7Hz,2H),8.03(s,1H),7.81(s,1H),7.76(d,J=6.7Hz,2H),7.10-7.03(m,1H),7.01-6.93(m,1H),6 .93-6.84(m,1H),5.01(s,2H),3.50(q,J=7.6Hz,1H),3.12-2.99(m,2H),2.92-2.77(m,2H),2.25-2.15(m,2H),1.20(d,J=7.0Hz,3H).

[0256] 13 C NMR(101MHz,CDCl3)δ161.28,158.76(d,J=243.8Hz),153.95(d,J=238.8Hz),1 51.08,143.86,141.82,129.76(dd,J=15.8,7.1Hz),126.63,116.95(dd,J=26. 9,8.2Hz),116.57(dd,J=24.4,9.2Hz),115.94(dd,J=26.3,4.6Hz),78.35(d,J =5.0Hz), 56.32 (d, J = 6.5Hz), 54.60 (d, J = 3.8Hz), 37.96, 34.17, 28.43, 17.09.

[0257] Example 28, Preparation of Compound L-27

[0258] (2R,3R)-2-(2,5-difluorophenyl)-3-((4-(pyridin-4-yl)butyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0259]

[0260] Intermediate II (200 mg, 0.507 mmol, 1.0 equiv.) and (2,4-dichlorophenyl)methanethiol (3.0 equiv.) were reacted according to Example 3, and the target product L-27 (92 mg) was obtained by column chromatography.

[0261] 1H NMR (400MHz, CDCl3) δ8.78(d,J=6.6Hz,2H),8.03(s,1H),7.79(s,1H),7.75(d,J=6.7Hz,2H),7.10-7.03(m,1H),7.01-6.93(m,1H),6 .92-6.85(m,1H),5.01(s,2H),3.55-3.45(m,1H),2.95(t,J=7.4Hz,2H),2.89-2.77(m,2H),1.95-1.78(m,4H),1.20(d,J=7.1Hz,3H).

[0262] 13 C NMR (101MHz, CDCl3) δ161.99,160.10-157.36(m),155.30-152.64(m),151.06,143.9 0,141.61(d,J=2.9Hz),129.89(dd,J=15.5,7.0Hz),126.56(d,J=1.9Hz),116.67(dd, J=71.2,8.9Hz),116.66(dd,J=20.0,8.8Hz),115.88(dd,J=26.3,4.7Hz),78.36(d,J =5.2Hz), 56.36 (d, J = 6.0Hz), 54.74 (d, J = 3.7Hz), 38.79, 35.48, 28.10, 28.05, 17.09.

[0263] Example 29: Preparation of deuterated compound L-28

[0264] (2R,3R)-2-(2,5-difluorophenyl)-3-((2-(pyridin-4-yl)ethyl-2-d)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0265]

[0266] Thioacetic acid (1.43 mL, 20.0 mmol, 2.0 equiv.) and deuterated water (3 mL) were mixed and stirred for 30 minutes, and then a solution of 4-vinylpyridine (1.08 mL, 10.0 mmol, 1.0 equiv.) and DTBP (3.67 mL, 20 mmol, 1.5 equiv.) in DCM (10 mL) was added to the system. The mixture was stirred at room temperature overnight under light, and DCM (20 mL) and water (10 mL) were added for separation. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was spin-dried and purified by column chromatography to give a colorless liquid S-(2-(pyridin-4-yl)ethyl-2-d)ethanethioate (1.40 g, yield: 77%).

[0267] The above colorless liquid (546 mg, 3.0 mmol, 1.0 equiv.) was dissolved in MeOH (10 mL). Under nitrogen protection, potassium carbonate (828 mg, 6.0 mmol, 2.0 equiv.) was added at room temperature. After the system was stirred at room temperature for 2 hours, water (20 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (3*40 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried. The mixture was purified by column chromatography to obtain a light yellow liquid 2-(pyridin-4-yl)ethane-2-d-1-thiol (230 mg, yield: 55%).

[0268] 1 H NMR (400MHz, CDCl3): δ8.41 (d, J = 5.4Hz, 2H), 7.08-6.95 (m, 2H), 2.78 (q, J = 6.6, 5.9Hz, 1H), 2.68 (q, J = 7.2Hz, 2H), 1.31 (t, J = 8.0Hz, 1H).

[0269]

[0270] Compound II (300 mg, 0.761 mmol, 1.0 equiv.) and the above compound (1.2 equiv.) were dissolved in methanol (10 mL) and stirred at room temperature for 12 hours. After the reaction was completed, the solvent was removed by vortexing and purified by column chromatography to obtain a white solid L-28 (yield: 52%). HRMS (ESI) calculation for C 19 H 20 DF2N4OS2[M+H] + m / z=424.1182,found:424.11823.

[0271] Example 30: Preparation of Compound L-29

[0272] (2R,3R)-3-((2,2-difluoro-2-phenylethyl)disulfanyl)-2-(2,5-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0273]

[0274] Compound II (300 mg, 0.761 mmol, 1.0 equiv.) and 2,2-difluoro-2-phenylethane-1-thiol (1.5 equiv.) were dissolved in methanol (20 mL) and stirred at room temperature overnight. After the reaction was completed, the solvent was removed by vortexing and purified by column chromatography to obtain a colorless viscous liquid L-29 (192 mg). HRMS (ESI) calculation for C 20 H 19 F4N3NaOS2 + [M+Na] + m / z=480.0798,found:480.0802.

[0275] Example 31, Preparation of Compound L-30

[0276] (2R,3R)-2-(2,5-difluorophenyl)-3-(methylthio)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0277]

[0278] Compound L-1 (1.0 mmol) was dissolved in DMF (5 mL), NaH (1.5 mmol) was added, and the mixture was stirred at zero degree for 5 min, followed by addition of iodomethane (1.5 mmol), and stirred at room temperature for half an hour. After purification by column chromatography, L-30 was obtained, a colorless viscous liquid with a yield of 56%.

[0279] 1 H NMR(400MHz, CDCl3)δ7.83(s,1H),7.79(s,1H),7.15-7.06(m,1H),6.97-6.90(m,1H),6.90 -6.83(m,1H),5.12-4.88(m,2H),3.24(q,J=7.0Hz,1H),2.27(s,3H),1.17(d,J=7.0Hz,3H).

[0280] 13C NMR (101MHz, CDCl3) δ157.86 (d, J = 243.5Hz), 153.65 (d, J = 235.1Hz), 151.24, 143.61, 129.92 (d, J = 15.3Hz), 117 .09-116.28(m),116.67(dd,J=8.8,3.5Hz),116.35-116.05(m),79.36,60.38,56.44(d,J=6.5Hz),47.00,16.20.

[0281] Example 32: Preparation of Compound L-31

[0282] (2R,3R)-2-(2,5-difluorophenyl)-3-(methylsulfinyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0283]

[0284] Compound L-30 (2 mmol) was dissolved in DCM (10 mL), and PhIO (2.4 mmol) was added, and stirred at room temperature for 30 minutes. After the reaction was completed, the solvent was removed by vortexing, and L-31 was obtained after purification by column chromatography (yield: 82%). HRMS (ESI) calculation for C 13 H 16 F2N3O2S[M+H] + m / z=316.0926,found:316.0931.

[0285] Example 33. Preparation of Compound L-32

[0286] (2R,3R)-2-(2,5-difluorophenyl)-3-(methylsulfonyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0287]

[0288] Compound L-30 (2 mmol) was dissolved in DCM (10 mL), and PhIO (5.0 mmol) was added, and stirred at room temperature for 30 minutes. After the reaction was completed, the solvent was removed by vortexing, and L-32 was obtained after column chromatography purification (yield: 73%). HRMS (ESI) calculation for C 13 H 16 F2N3O3S[M+H] + m / z=332.0875,found:332.0881.

[0289] Comparative Example 1: Preparation of Compound M-1

[0290] (2R,3R)-2-(2,4-difluorophenyl)-3-(((1-methyl-1H-imidazol-2-yl)methyl)disulfanyl)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0291]

[0292] Compound L-1 (285 mg, 1.0 mmol, 1.0 equiv.), dibromomethane (1.5 equiv.) and 2-(pyridin-4-yl)ethane-1-thiol (1.5 equiv.) were dissolved in acetonitrile (10 mL), and DBU (2.0 equiv.) was slowly added dropwise at 0°C. After the addition was completed, the temperature was naturally raised to room temperature for 5 hours. After the reaction was completed, the solvent was removed by vortexing, and a colorless viscous liquid (185 mg) was obtained after column chromatography purification. The product can be further purified by semi-preparative method (acetonitrile and 0.1% TFA water system). HRMS (ESI) calculated for C 20 H 23 F2N4OS2 + [M+H] + m / z=437.1276,found:437.1270.

[0293] Comparative Example 2: Preparation of Compound M-2

[0294] (2R,3R)-2-(2,5-difluorophenyl)-3-((((3-(pyridin-4-yl)propyl)thio)methyl)thio)-1-(1H-1,2,4-triazol-1-yl)butan-2-ol

[0295]

[0296] Compound L-1 (285 mg, 1.0 mmol, 1.0 equiv.), dibromomethane (1.5 equiv.) and 3-(pyridin-4-yl)propane-1-thiol (1.5 equiv.) were dissolved in acetonitrile (10 mL), and DBU (2.0 equiv.) was slowly added dropwise at 0°C. After the addition was completed, the temperature was naturally raised to room temperature for 5 hours. After the reaction was completed, the solvent was removed by vortexing, and a colorless viscous liquid (202 mg, yield: 45%) was obtained after column chromatography purification. The product can be further purified by semi-preparation (acetonitrile and 0.1% TFA water system).

[0297] 1 H NMR (400MHz, CDCl3): δ8.78(s,2H),8.16(s,1H),7.89(s,1H),7.75(d,J=5.6Hz,2H),7.35 (td,J=9.1,6.2Hz,1H),6.81-6.69(m,2H),5.06(d,J=14.0Hz,1H),4.91(d,J=14.1Hz,1H), 3.88(d,J=13.7Hz,1H), 3.78(d,J=13.7Hz,1H), 3.52(q,J=7.0Hz,1H), 3.05(dd,J=8.8,6. 6Hz,2H),2.87-2.68(m,2H),2.10(p,J=7.2Hz,2H),1.17(d,J=7.0Hz,3H).HRMS(ESI)calcd for C 21 H 25 F2N4OS2 + [M+H] + m / z=451.1432,found:451.1426.

[0298] Verification of the antifungal effect of compounds

[0299] 1. Fungal growth inhibition experiment:

[0300] 1: Strain activation

[0301] Take the Candida albicans strain frozen in -80℃ ultra-low temperature freezer and streak it on SDA plate, incubate at 30℃ for about 48h until a single colony grows. Store in 4℃ refrigerator for use within one week.

[0302] 2: Minimum inhibitory concentration (MIC) determination

[0303] Take the strain activated overnight, centrifuge and wash it three times with PBS, centrifuge (8000rpm, 1min), count it with a hemocytometer, and then adjust the concentration of the bacterial suspension to 0.5-2.5×10 with RPMI-1640 medium.3 CFU / mL. Take a sterile 96-well plate, add 100μL of blank RPMI-1640 liquid to well No. 12 in each row as a blank control; add 100μL of the adjusted bacterial suspension to each well No. 2 to No. 11; add 200μL of bacterial suspension and the drug to be tested to well No. 1; then use a multichannel pipette to dilute wells No. 2 to No. 10 by 2 times, well No. 11 does not contain drugs, and only 100μL of bacterial liquid is added as a growth control. The 96-well plate was cultured in a 30℃ / 35℃ constant temperature incubator for 24-48h, and the OD value of each well was measured at 630nm with an enzyme reader (measured for 24h and 48h), supplemented by visual inspection. The lowest concentration at which there is no bacterial growth observed by naked eye is the MIC value, and the lowest drug concentration corresponding to the inhibition of Candida albicans cell growth by more than 80% compared with the growth control well is calculated by the OD value, which is the MIC of the drug. 80 Among them, SC5314: sensitive Candida albicans.

[0304] Table 1: In vitro antibacterial activity (MIC) of some compounds 80 ,μg / ml)

[0305]

[0306]

[0307] From the above results, it can be seen that since the present invention adds a disulfide bond in the molecular design, it can form an effective π-S bond (Pi-S bond) interaction with the target. In addition, by introducing an aromatic group, especially a nitrogen-containing heterocycle, into the side chain, it can better bind to the enzyme target 14α-sterol demethylase through a variety of interaction forces, thereby obtaining excellent in vitro activity. The activity of some compounds is even 250 times higher than that of fluconazole. It should be pointed out that the in vitro antifungal activity of the control compounds M-1 and M-2 is similar to that of L-20, but the results of later animal experiments confirm that the efficacy of M-1 and M-2 is not as good as that of L-20.

[0308] Table 2: In vitro antibacterial activity (MIC 80 ,μg / ml)

[0309]

[0310]

[0311] As can be seen from Table 2, compound L-20 is effective against various Aspergillus such as Aspergillus flavus, Aspergillus niger and Aspergillus fumigatus. At the same time, compound L-20 is effective against various Candida such as Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei and Candida parapsilosis. In addition, compound L-20 is effective against Cryptococcus including Cryptococcus neoformans and Cryptococcus gattii. Many of the above strains are resistant bacteria isolated from recent clinical studies, and the effect of L-20 is significantly better than fluconazole.

[0312] Table 3: In vitro antibacterial activity (MIC) of compounds L-30 to L-32 80 ,μg / ml)

[0313]

[0314] In addition, we determined the activities of compounds L-30, L-31 and L-32 against Candida SC5314 and Aspergillus and found that the activities of these compounds were better than fluconazole.

[0315] 2. Animal Experimental Efficacy Verification of Compound L-20 and Comparative Examples M-1 and M-2:

[0316] The animal model experimental design is shown in Table 4 below:

[0317] C57 mice (male, 5 weeks old) were used. On day -1, cyclophosphamide 100 mg / kg (10 mg / ml×200 μL ip) was intraperitoneally injected. On day 0, Candida albicans 0304103 (2.5×10 6 CFU / mL×200μL iv), and then the positive drug fluconazole and the test compound were intraperitoneally injected for three consecutive days. The dose was 1mg / kg (0.1mg / ml×200μL ip). The mice were killed on the fourth day, and the antifungal activity of the compound was determined by calculating the bacterial load in the kidney.

[0318] Table 4

[0319]

[0320] Drug-resistant Candida albicans 0304103: (2.5×10 6 CFU / mL×200μL iv), solvent: PBS

[0321] Fluconazole: 1 mg / kg (0.1 mg / ml×200 μL ip) Solvent: 10% DMSO+40% PEG300+50% PBS

[0322] Test compound: 1 mg / kg (0.1 mg / ml×200 μL ip) Solvent: 10% DMSO+40% PE G300+50% PBS.

[0323] Animal experiment results Figure 1 As shown, it can be seen that in this Candida albicans animal model, there were animal deaths in the blank control group (mortality rate 50%), while no animals died in the drug-treated group. At the same time, the kidney bacterial load test found that compound L-20 was better than fluconazole (FKZ) and compounds M-1 and M-2, with significant statistical differences. Although the in vitro activity of compounds M-1 and M-2 is similar to that of L-20, they cannot be metabolized into L-1 and cannot exert a dual mechanism of action, so the animal experimental efficacy is not as good as L-20. This proves that the design concept of the present invention is effective.

[0324] 3. Animal Experimental Efficacy of Compounds L-30 and L-32 against Candida albicans

[0325] On day 0, Candida albicans SC5314 (2.5×10 6 CFU / mL×200μL iv), and then the positive drug fluconazole and the test compound were intraperitoneally injected at 0, 24, and 48 hours respectively. The dose was 10mg / kg (1.0mg / ml×200μL ip). The survival status of the mice was observed for ten consecutive days. The results showed that at this dose, the mice in the fluconazole group, L-30 and L-32 groups could survive.

[0326] Table 5

[0327] Compound Average survival Fluconazole 10 L-30 10 L-32 10

[0328] 4. Animal Experimental Efficacy of Compounds L-30 and L-32 against Aspergillus

[0329] On day 0, the cells were injected with Aspergillus MTU6001 (2.5 × 10 6 CFU / mL×200μL iv), and then the positive drug fluconazole and the test compounds L-30 and L-32 were intraperitoneally injected at 0, 24, and 48 hours respectively. The dose was 50mg / kg (5.0mg / ml×200μL ip). The survival status of the mice was observed for ten consecutive days. The results are shown in Table 6 below. The survival period of mice in the L-30 and L-32 groups was significantly longer than that in the fluconazole group.

[0330] Table 6

[0331] Compound Average survival Fluconazole 2.8 L-30 9.8 L-32 10

[0332] 5. PK experiment:

[0333] C57 mice (male, 5 weeks old) were intraperitoneally injected with L-20 (10 mg / kg, 200 μL). 25 ul of mouse plasma was taken at 5 and 60 min, and an aqueous solution of dithiothreitol DTT (0.2 mg / ml, 50 ul) was added, and vortexed at 20 degrees for 60 minutes. Then 100 ul of acetonitrile was added, centrifuged, and the upper layer of liquid was sent for mass spectrometry analysis to monitor the main metabolite L-1. The data obtained based on the peak area are shown in Table 7 below:

[0334] Table 7

[0335] Entry Sample Name Peak area of ​​L-1 1 5min-without DTT Below detection limit 2 5min-Add DTT 11760 3 60min-without DTT Below detection limit 4 60min-Add DTT 2629

[0336] The above results show that at the 5th and 60th minutes, the peak area of ​​L-1 in the free state is very small, below the detection limit (Entry 1 and Entry 3). However, after adding DTT for reduction, a large amount of free L-1 can be detected (Entry 2 and Entry 4). This shows that L-1 exists in plasma mainly in the form of disulfide bonds with endogenous cysteine, and very little is free. Similar phenomena have been reported in many literatures (Drug metabolism and disposition, 2006, 34, 961; Drugmetabolism and disposition, 2001, 29, 60). Therefore, it can be reasonably inferred that the metabolite L-1 of this series of compounds can also form disulfide bonds with cysteine-containing enzymes in fungi, thereby inhibiting the activity of fungal-related enzymes. The large amount of glutathione in the human body can neutralize this effect and protect the human body from this effect (see reference: The mode of action of allicin: trapping of radicals and interaction with thiol-containing proteins, Biochimica et Biophysica Acta, 1998, 1379, 233). At the same time, this type of antifungal drug can also inhibit fungal 14α-sterol demethylase like traditional conazole drugs, thereby exerting a dual mechanism of action.

[0337] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art may make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A sulfur-containing triazole compound and its salt, wherein the compound has the following general formula I: in, A is selected from C1-C6 alkyl, C1-C6 fluoroalkyl, or a group in which one or more atoms of C1-C6 alkyl or C1-C6 fluoroalkyl are substituted by their isotopes; Ar represents a substituted or unsubstituted 5-14 membered aryl or heteroaryl group; the heteroatom on the heteroaryl group is selected from N, S, and O; and the substituent is selected from a halogen atom or a C1-C6 alkyl group.

2. The sulfur-containing triazole compound and its salt according to claim 1, characterized in that: A is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, difluoroethyl or isotope-substituted ethyl.

3. The sulfur-containing triazole compound and its salt according to claim 1, characterized in that: Ar is selected from one of the following groups: phenyl, halophenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, isoxazolyl, imidazolyl, thiazolyl, furanyl, pyrazolyl, oxazolyl, pyrrolyl, indolyl, isoindolyl, quinolyl, isoquinolyl.

4. A sulfur-containing triazole compound and its salt, wherein the compound has one of the following structural formulas: in, R is a hydrogen atom or a C1-C6 alkyl group.

5. The sulfur-containing triazole compound and its salt according to claim 1 or 4, characterized in that: The salt is selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, formate, acetate, benzoate, benzenesulfonate, camphorsulfonate, citrate, edisylate, fumarate, glucoheptonate, gluconate, glucuronate, isethionate, lactate, lactobionate, dodecyl sulfate, malate, maleate, methanesulfonate, naphthoate, naphthoate, nitrate, stearate, oleate, oxalate, pamoate, phosphate, hydrogen phosphate, dihydrogen phosphate, polygalacturonate, succinate, sulfate, sulfosalicylate, tartrate, p-toluenesulfonate, and trifluoroacetate.

6. A method for preparing sulfur-containing triazole compounds and salts thereof according to claim 1, characterized in that: The method comprises the following steps: S1, thiol compound L-1 Reaction with disulfide compound R1-SS-R1 to generate intermediate II wherein R1 is selected from an electron withdrawing group; S2. The intermediate II reacts with thiol Ar-A-SH to obtain the sulfur-containing triazole compound.

7. The method for preparing sulfur-containing triazole compounds and salts thereof according to claim 6, characterized in that: The disulfide compound is selected from 2,2'-disulfide dipyridine, 4,4'-disulfide dipyridine, 3,3'-disulfide dipyridine, 1,2-di(pyrimidin-2-yl)disulfane, and 4,4'-dinitrodiphenyl disulfide.

8. The method for preparing the sulfur-containing triazole compound and its salt according to claim 6, characterized in that: The solvent used in the reactions of steps S1 and S2 is selected from at least one of methanol, ethanol, ethyl acetate, tetrahydrofuran, DMSO, dichloromethane, chloroform and acetonitrile.

9. An isotope-labeled compound of a thiotriazole compound and a salt thereof according to claim 1 or 4, characterized in that: The isotope-labeled compound has the structure of the thiotriazole compound, and one or more atoms in the structural formula are replaced by its isotope.

10. Use of the sulfur-containing triazole compound and its salt according to claim 1, 4 or 9 in the preparation of antifungal drugs.

11. The use according to claim 10, characterized in that The antifungal drug comprises at least one of an anti-Candida drug, an anti-cryptococcal drug, and an anti-Aspergillus drug.

12. A pharmaceutical composition comprising at least one sulfur-containing triazole compound and its salt, or its isomer according to claim 1, 4 or 9, and a pharmaceutically acceptable carrier, diluent or excipient.

Images