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WO2025011553A1 - Azole compounds, synthesis therefor and pharmaceutical use thereof - Google Patents

Azole compounds, synthesis therefor and pharmaceutical use thereof Download PDF

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Publication number
WO2025011553A1
WO2025011553A1 PCT/CN2024/104464 CN2024104464W WO2025011553A1 WO 2025011553 A1 WO2025011553 A1 WO 2025011553A1 CN 2024104464 W CN2024104464 W CN 2024104464W WO 2025011553 A1 WO2025011553 A1 WO 2025011553A1
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alkyl
substituted
halogen
compound
nmr
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French (fr)
Chinese (zh)
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梁晓霞
王雅璐
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Sichuan Agricultural University
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Sichuan Agricultural University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C07D233/58—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring nitrogen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C07D233/60—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with hydrocarbon radicals, substituted by oxygen or sulfur atoms, attached to ring nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
    • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles

Definitions

  • the C 1 -C 3 alkoxy group refers to: methoxy group, ethoxy group, n-propyloxy group, isopropyloxy group.
  • the C 2 -C 4 alkenyloxy group refers to vinyloxy, propenyloxy, butenyloxy and the like.
  • reaction mixture was cooled to room temperature, and the crude product was separated by column chromatography to obtain the cyano-substituted compound 18 of compound 4, i.e., compound 18, the structure of which was confirmed by 1 H NMR, 13 C NMR and HR-MS.
  • the reaction was carried out in a continuous flow jet bubbling reactor with a glass tube (8 mm inner diameter) at atmospheric pressure.
  • the alkyl substituted compound 20 of compound 4, i.e., compound 20, was obtained, and its structure was confirmed by 1 HNMR, 13 C NMR and HR-MS.
  • test compound 20 1 H NMR, 13 C NMR and HR-MS, the results of test compound 20 are as follows:
  • Fluconazole, ketoconazole, itraconazole and the synthesized compound were dried under reduced pressure at 25°C for 12h, the dried samples were accurately weighed, and an appropriate amount of 20% DMSO was added to dissolve, and the volume was fixed to prepare 4mL of stock solutions with drug molar concentrations of 6 ⁇ mol/mL, 3 ⁇ mol/mL, 1.5 ⁇ mol/mL, 0.75 ⁇ mol/mL, and 0.375 ⁇ mol/mL, respectively. After filtering with a 0.22 ⁇ m microporous filter membrane, the samples were stored in a refrigerator at 4°C for later use.
  • PDA potato agar medium
  • the colony diameter of the test fungi on the drug-containing culture medium was measured by the cross method, and the inhibition rate of each drug solution treatment on the linear growth of mycelium was calculated by comparing with the control.
  • the 1st column is a blank control
  • the 11th column is a negative control
  • the 12th column is a growth control.
  • the 96-well plate was placed in a 27°C constant temperature incubator for 48 hours, and the minimum drug concentration of the well with clear and transparent solution was the MIC. Three groups of replicates were set for each.
  • compound 5 had a very obvious inhibitory effect on the hyphal growth of Microsporum canis, and compounds 3 and 4 had the greatest influence on the hyphal growth rate of Trichophyton mentagrophytes and Microsporum gypseum, both less than 1 mm/d.
  • ketoconazole had the highest antibacterial rate, and its antibacterial effect was better than that of itraconazole and fluconazole.
  • Compounds 1-8 had different degrees of antibacterial effect on the three fungi. At a concentration of 6 ⁇ mol/mL, the antibacterial rates of compounds 3 and 4 on the three fungi were greater than 60%, with good antibacterial effects.
  • the antibacterial rate of compound 2 on Microsporum canis and Trichophyton mentagrophytes was greater than 60%, with good antibacterial effects, but the antibacterial rate for Microsporum gypseum was greater than 45%, with good antibacterial effects.
  • Compound 1 had a good antibacterial effect on Trichophyton mentagrophytes, and had a good antibacterial effect on Microsporum canis and Microsporum gypseum.
  • compound 4 still had a good antibacterial effect on the three fungi
  • compound 3 had a good antibacterial effect on Trichophyton mentagrophytes and Microsporum gypseum, but had a poor antibacterial effect on Microsporum canis
  • compound 1 had a good antibacterial effect.
  • the antifungal test showed that, among the 20 compounds, except for compounds 5-8, the other compounds had antifungal activities against the four pathogenic fungi that were superior to or equivalent to the positive control.
  • the compounds 1-20 synthesized in the above examples were subjected to normal cell cytotoxicity assay.
  • NIH3T3 cells grow well and are in the logarithmic growth phase, follow the subculture procedure, resuspend the cells in 1 mL of complete culture medium, add an appropriate amount of complete culture medium to dilute, and count to 5 ⁇ 10 4 cells/mL. Take the prepared cell suspension and inoculate it into a 96-well cell culture plate, add 100 ⁇ L to each well, seal it, and transfer it to a 37°C, 5% CO 2 constant temperature incubator for 24 hours.
  • the compound was dissolved in DMSO, and the corresponding solvent was used as the negative control group. 100 ⁇ L of complete medium was added to the control group, and the complete medium was supplemented to 200 ⁇ L in the experimental group, so that the final concentration of the drug solution in each well was 20 ⁇ M, and 5 groups were paralleled. Derivatives that showed good inhibitory activity at a concentration of 20 ⁇ M were rescreened for active drugs using NIH3T3 cells according to a certain concentration gradient.
  • the MTS solution and complete culture medium were mixed at a ratio of 1:10, the medium containing the drug solution in the 96-well plate was discarded, 200 ⁇ L of the mixed solution was added to each well, and 5 blank groups of the mixed solution without cells were set up and placed in a constant temperature box for reaction for 2-4 hours.
  • the absorbance of each well at an absorption wavelength of 490 nm was detected by an ELISA instrument, the data were exported and the inhibition rate was calculated, and the IC 50 was calculated by the Bliss method using GraphPad Prism 8 software.
  • compound 3 has the smallest IC 50 and the greatest cytotoxicity, while the remaining compounds, including ketoconazole, have lower cytotoxicity. This shows that the toxicity of these compounds to normal tissues should be lower than that of ketoconazole.
  • the positive controls fluconazole and itraconazole have no obvious inhibitory effect on Microsporum canis, Trichophyton mentagrophytes, and Microsporum gypseum, while ketoconazole has a significant inhibitory effect on these three bacteria.
  • the three positive controls all have a certain inhibitory effect on Candida albicans.
  • the synthesized compounds 1-20 have different degrees of antibacterial effects on Microsporum canis, Trichophyton mentagrophytes, Microsporum gypseum, and Candida albicans.
  • compounds 4, 9, 10, 13, 14, 16, and 17 have obvious inhibitory effects on these four bacteria, and are better than fluconazole, itraconazole, and ketoconazole (partially equivalent); compounds 11, 12, and 18 are second; compounds 1-3, 15, 19, and 20 are third. And compounds 1-20 show low cytotoxicity to normal cells and have high safety. The above compounds all have potential antifungal application value.
  • Representative compounds 1-4 were selected, and the molecular docking method was used to examine the binding and action mode of the representative compounds with common antifungal target proteins, and to predict the possible targets of the active compounds in this application.
  • sterol 14- ⁇ demethylase (CYP51, PDB ID: 5FSA) of pathogenic yeast Candida albicans; sterol 14- ⁇ demethylase (CYP51, PDB ID: 5TZ1) of Candida albicans; exo- ⁇ (1,3)-glucanase (XOG1, PDB ID: 1EQP) of Candida albicans; Exo- ⁇ (1,3)-glucanase (XOG1, PDB ID: 1CZ1); chitin synthase 2 from Candida albicans (CHS2, PDB ID: 7STL); dihydrofolate reductase from Candida albicans (DFR1, PDB ID: 1AI9); Candida albicans N-myristoyltransferase (NMT1, PDB ID: 1IYK); phosphomannose isomerase from Candida albicans (PMI, PDB ID: 1PMI);
  • Lib Dock Score indicates the degree of binding between the ligand and the target protein crystal. The higher the Lib Dock Score, the higher the activity of the small molecule binding to the receptor.
  • Compounds 1-4 were molecularly docked with the proteins in "1.1" and visualized. Representative binding diagrams are shown in Figures 1 to 4, which respectively show the spatial configuration of compound 4 molecularly docked with 1EQP, 1CZ1, 1AI9, and 4IIB, as well as the binding of the active groups of the compounds.
  • Lib Dock Score is shown in Table 9.
  • Sterol 14- ⁇ demethylase is a potential antifungal target for most azole compounds, but molecular docking revealed that, compared with the most active ketoconazole, compounds 1-4 had weak binding ability to sterol 14- ⁇ demethylase (CYP51, PDB ID: 5FSA and CYP51, PDB ID: 5TZ1), but showed higher binding ability to glucanase (XOG1, PDB ID: 1EQP, 1CZ1 and XOG1, PDB ID: 1CZ1), dihydrofolate reductase (DFR1, PDB ID: 1AI9) and ⁇ -glucosidase 1 (BGL1, PDB ID: 4IIB).
  • the molecular docking results show that the synthesized active compounds are different from the targets of traditional azole compounds. Its antifungal effect may be highly related to glucanase, dihydrofolate reductase and ⁇ -glucosidase 1.
  • a fur animal topical spray (10 mg/mL) is prepared from the following raw materials by weight percentage: 1% active compound, 40% ethanol, 5% glycerol, 0.2% sodium thiosulfate, 0.5% water-soluble azone, 0.2% sorbic acid, 0.5% PVA, 2% PVP, and 1% Tween 80.
  • the active compound can be any one of the compounds 1-20 prepared in the above embodiments.
  • the preparation method is as follows: take various raw material ingredients according to the above raw material formula ratio, put alcohol in a beaker, add glycerol, sodium thiosulfate, water-soluble azone, sorbic acid, stir for 10 minutes, and mix evenly. Then slowly add PVA (polyvinyl alcohol, Kuraray PVA-217, the same below) and PVP (polyvinyl pyrrolidone, k30, the same below), stir well, and heat appropriately to promote dissolution. After PVA and PVP are fully dissolved, add Tween 80 and continue stirring for 20 minutes. Cool to below 50°C, add active compounds, stir until fully dissolved, filter, and bottle to obtain a fur animal external spray.
  • PVA polyvinyl alcohol
  • PVP polyvinyl pyrrolidone
  • a fur animal topical spray (10 mg/mL) is prepared from the following raw materials by weight percentage: 1% active compound, 5% DMSO, 10% Tween 80, 20% PEG400, 20% ethanol, 5% glycerol, 0.2% sodium thiosulfate, 0.5% water-soluble azone, 0.2% sorbic acid, 0.5% PVA, and 2% PVP.
  • the active compound can be any one of the compounds 1-20 prepared in the above embodiments.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Communicable Diseases (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Provided are azole compounds, synthesis therefor and the pharmaceutical use thereof. The azole compounds have molecular structures represented by formula I, wherein at least one R1 is present; the R1 is each independently selected from hydrogen, C1-C4 alkyl, cyclohexyl, halogen-substituted C1-C3 alkyl, C2-C4 alkenyl, halogen-substituted C2-C4 alkenyl, nitro, amino, halogen, methylamino, hydroxyl, carboxyl, hydroxyl-substituted C1-C3 alkyl, phenyl, methylphenyl, benzyl, C1-C3 alkoxy, C2-C4 alkenyloxy, sulfo, nitrophenyl, formyl, acetyl, benzoyl, benzyloxycarbonyl, benzyloxy, phenoxy, formyloxy, acetoxy, benzoyloxy, mercapto, methylsulfonyl, sulfo, C1-C3 alkylamino, bi-C1-C3 alkyl-substituted amino and tert-butoxycarbonyl; a=0-4; b=0-4; at least one R2 is present; the R2 is each independently selected from C1-C3 alkyl, halogen, amino, methylamino, dimethylamino, hydroxyl, carboxyl, nitro, cyano and haloalkyl; X1 is -(CH2)n- or -(CH2)n-O- or -O-(CH2)n-, n=0-4; X2 is -(CH2)m- or -C(O)-(CH2)m-, m=0-4; and Y is C or N.

Description

一种唑类化合物及其合成、制药应用Azole compound and its synthesis and pharmaceutical application

本申请要求了2023年7月10日提交的中国专利申请2023108400143的优先权。This application claims priority to Chinese patent application 2023108400143 filed on July 10, 2023.

技术领域Technical Field

本发明涉及杂环化合物领域，特别是唑类抗真菌化合物及其制备方法，属于药物化合物的合成领域。The invention relates to the field of heterocyclic compounds, in particular to azole antifungal compounds and preparation methods thereof, and belongs to the field of synthesis of pharmaceutical compounds.

背景技术Background Art

真菌感染是人和动物共患疾病。人类致病真菌常见念珠菌、癣菌、隐球菌等；毛皮动物真菌性皮肤病是临床上常见的动物疾病，病原主要为犬小孢子菌(Microsporum canis)、石膏样小孢子菌(Microsporum gypseum)、须癣毛癣菌(Trichophyton mentagrophytes)和疣状毛癣菌(Trichophyton verrucosum)。感染后会引起人、牛、马、猪、猫、犬等的皮肤癣病，且感染较为顽固，不易根治，容易出现交叉感染和传播的情况，若不及时治疗可能会诱发全身感染，危害性较大。Fungal infection is a disease common to humans and animals. Common pathogenic fungi in humans include Candida, ringworm, Cryptococcus, etc. Fungal skin diseases in fur animals are common animal diseases in clinical practice, and the main pathogens are Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes, and Trichophyton verrucosum. After infection, it can cause skin ringworm in humans, cattle, horses, pigs, cats, dogs, etc., and the infection is relatively stubborn and difficult to cure. It is easy to cross-infect and spread. If not treated in time, it may induce systemic infection, which is very harmful.

真菌感染治疗所需周期长、过程复杂、成本高，人医临床常用酮康唑、伊曲康唑、氟康唑，但已出现明显耐药；兽医临床上常用人医抗真菌药特比萘酚、酮康唑、伊曲康唑进行治疗，缺乏动物专用抗真菌化合物。一方面与《兽药管理条例》四十一条关于“禁止将人用药品用于动物”的规定相冲突，另一方面临床所用的抗真菌药物多存在真菌耐药性、药物不良反应、抗菌谱窄、组织浓度低、口服不吸收等问题。导致大量的抗真菌药物不恰当使用，逐渐显示出药物滥用问题，严重危害药物正常应用开发。The treatment of fungal infections requires a long cycle, a complex process, and high costs. Ketoconazole, itraconazole, and fluconazole are commonly used in human clinical practice, but obvious drug resistance has emerged; terbinafine, ketoconazole, and itraconazole are commonly used in veterinary clinical practice for treatment, and there is a lack of animal-specific antifungal compounds. On the one hand, it conflicts with Article 41 of the "Regulations on Veterinary Drug Administration" on "prohibiting the use of human drugs in animals". On the other hand, most antifungal drugs used in clinical practice have fungal resistance, adverse drug reactions, narrow antibacterial spectrum, low tissue concentration, and oral non-absorption. This has led to the inappropriate use of a large number of antifungal drugs, which have gradually shown drug abuse problems and seriously endangered the normal application and development of drugs.

故开发安全性高、抗耐药、使用方便的抗真菌药物，特别是新型动物专用抗真菌药，很有必要。特别是，如何实现高效抗真菌、低毒副作用，对于养殖畜牧业具有重要意义，如果能够开发一种有效治疗动物真菌感染的药物，可以大幅度减少养殖业因为动物感染真菌导致的生长缓慢、动物死亡等问题，使得养殖业的成本降低，经济效益提高。Therefore, it is necessary to develop antifungal drugs with high safety, resistance to drug resistance, and convenience of use, especially new antifungal drugs for animals. In particular, how to achieve high antifungal efficiency and low toxicity is of great significance to animal husbandry. If a drug that can effectively treat animal fungal infections can be developed, it can greatly reduce the problems of slow growth and animal death caused by fungal infections in animals in the breeding industry, thereby reducing the cost of the breeding industry and improving economic benefits.

发明内容Summary of the invention

本发明的目的在于克服现有技术中所存在的缺少动物专用抗真菌药物的不足，且现有人的医用药应用于动物抗真菌治疗也存在耐药性和不良反应的现状，提供一种新型氮唑类化合物。该类化合物可以有效抑制犬小孢子菌、石膏样小孢子菌、须癣毛癣菌及白色念珠菌，且毒性较低，可应用于抗真菌应用，特别是兽医临床中抗真菌应用。 The purpose of the present invention is to overcome the shortcomings of the prior art, which lacks antifungal drugs specifically for animals, and the current situation that the application of human medical drugs in animal antifungal treatment also has drug resistance and adverse reactions, and to provide a new type of nitrogen azole compound. The compound can effectively inhibit Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes and Candida albicans, and has low toxicity, and can be used for antifungal applications, especially antifungal applications in veterinary clinics.

为了实现上述发明目的，本发明提供了以下技术方案：In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

一种唑类化合物，具有式I所示分子结构的化合物：
An azole compound having a molecular structure shown in Formula I:

其中，R1至少一个，R1各自独立的选自氢、C1-C4烷基、环己基、卤取代C1-C3烷基、C2-C4烯基、卤取代C2-C4烯基、硝基、氰基、氨基、卤素、甲氨基、羟基、羧基、羟基取代C1-C3烷基、苯基、甲基苯基、苯甲基、C1-C3烷氧基、C2-C4烯基氧基、磺酸基、硝基苯基、甲酰基、乙酰基、苯甲酰基、苄氧羰基、苄氧基、苯氧基、甲酰氧基、乙酰氧基、苯甲酰氧基、巯基、甲基磺酰基、磺酸基、C1-C3烷基氨基、双C1-C3烷基取代氨基或叔丁氧羰基。wherein, there is at least one R1 , and each R1 is independently selected from hydrogen, C1 - C4 alkyl, cyclohexyl, halogen-substituted C1- C3 alkyl, C2 - C4 alkenyl, halogen-substituted C2 - C4 alkenyl, nitro, cyano, amino, halogen, methylamino, hydroxyl, carboxyl, hydroxyl-substituted C1 - C3 alkyl, phenyl, methylphenyl, benzyl, C1 - C3 alkoxy, C2 - C4 alkenyloxy, sulfonic acid, nitrophenyl, formyl, acetyl, benzoyl, benzyloxycarbonyl, benzyloxy, phenoxy, formyloxy, acetoxy, benzoyloxy, mercapto, methylsulfonyl, sulfonic acid, C1 - C3 alkylamino, di- C1 - C3 alkyl-substituted amino or tert-butyloxycarbonyl.

a表示R1的个数，a＝0～4；b表示R2的个数，b＝0～4。a represents the number of R1 , a=0-4; b represents the number of R2 , b=0-4.

R2至少一个，R2各自独立的选自C1-C3烷基、卤素、氨基、甲氨基、二甲氨基、羟基、羧基、硝基、氰基或卤代烷基。There is at least one R 2 , and each R 2 is independently selected from C 1 -C 3 alkyl, halogen, amino, methylamino, dimethylamino, hydroxyl, carboxyl, nitro, cyano or halogenated alkyl.

X1为-(CH2)n-或-(CH2)n-O-或-O-(CH2)n-，n＝0～4。 X1 is -( CH2 )n- or -( CH2 )nO- or -O-( CH2 )n-, and n=0-4.

X2为-(CH2)m-或-C(O)-CH(CH3)-(CH2)m-，m＝0～4。X 2 is -(CH 2 )m- or -C(O)-CH(CH 3 )-(CH 2 )m-, m=0 to 4.

Y为C或N。Y is C or N.

进一步，所述R1各自独立的选自：氢、甲基、卤取代甲基、乙基、正丙基、异丙基、异丁基、烯丙基、丙炔基、乙烯基、仲丁基、环己基、苯甲基、丙烯基、叔丁基、异丙烯基、乙炔基、苯基、对甲苯基、间甲苯基、邻甲苯基、邻硝基苯基、甲酰基、乙酰基、苯甲酰基、羧基、苄氧羰基、氰基、氨基、甲氨基、乙基氨基、丙基氨基、二甲氨基、二乙氨基、硝基、羟基、甲氧基、乙氧基、苄氧基、苯氧基、甲酰氧基、巯基、氟基、氯基、溴基、碘基、叔丁氧羰基或氟甲基。Further, each of the R1s is independently selected from the group consisting of hydrogen, methyl, halogen-substituted methyl, ethyl, n-propyl, isopropyl, isobutyl, allyl, propynyl, vinyl, sec-butyl, cyclohexyl, benzyl, propenyl, tert-butyl, isopropenyl, ethynyl, phenyl, p-tolyl, m-tolyl, o-tolyl, o-nitrophenyl, formyl, acetyl, benzoyl, carboxyl, benzyloxycarbonyl, cyano, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, nitro, hydroxyl, methoxy, ethoxy, benzyloxy, phenoxy, formyloxy, mercapto, fluoro, chloro, bromo, iodo, tert-butyloxycarbonyl or fluoromethyl.

优选地，所述R1各自独立的选自：氢、羟基、氰基、甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、氟、氯、溴、三氟甲基、全氟乙基、氨基或叔丁氧羰基等。Preferably, the R 1 is independently selected from hydrogen, hydroxyl, cyano, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, fluorine, chlorine, bromine, trifluoromethyl, perfluoroethyl, amino or tert-butyloxycarbonyl, etc.

进一步，所述R2各自独立的选自：C1-C3烷基、卤素、氨基、硝基、氰基、氟基、氯基、溴基、碘基、羟基、全氟C1-C3烷基。全氟C1-C3烷基包括全氟甲基、全氟乙基、全氟正丙基或全氟异丙基。Furthermore, each of the R2 groups is independently selected from: C1- C3 alkyl, halogen, amino, nitro, cyano, fluoro, chloro, bromo, iodo, hydroxyl, perfluoro C1 - C3 alkyl. Perfluoro C1 - C3 alkyl includes perfluoromethyl, perfluoroethyl, perfluoro-n-propyl or perfluoro-isopropyl.

进一步，X1和X2处于苯环的邻位或对位。X1和X2在共同连接的苯环上，分别处于该苯环的邻位或对位。 Furthermore, X1 and X2 are located at the ortho position or para position of the benzene ring. X1 and X2 are located at the ortho position or para position of the benzene ring on the benzene ring to which they are connected.

进一步，a表示R1的个数，a＝0～3。优选地，a＝0、1、2。Furthermore, a represents the number of R 1 , and a=0 to 3. Preferably, a=0, 1, 2.

进一步，b表示R2的个数，b＝0～3。优选地，b＝0、1、2。Furthermore, b represents the number of R 2 , and b = 0 to 3. Preferably, b = 0, 1, 2.

进一步，X1选自亚甲基、亚乙基、亚甲氧基或亚乙氧基。可以是亚甲基或亚乙基，也可以是亚甲氧基、亚乙氧基，亚乙氧基是指-CH2-CH2-O-。Furthermore, X1 is selected from methylene, ethylene, methyleneoxy or ethyleneoxy. It may be methylene or ethylene, or methyleneoxy or ethyleneoxy. Ethyleneoxy means -CH2 - CH2 -O-.

进一步，X2选自-C(O)-CH(CH2)-、亚甲基或亚乙基。Further, X 2 is selected from -C(O)-CH(CH 2 )-, methylene or ethylene.

进一步，n＝0～3。优选地，n＝0、1、2。Furthermore, n=0 to 3. Preferably, n=0, 1, 2.

进一步，m＝0～3。优选地，m＝0、1、2。例如，m＝0的时候，X2可以为-C(O)-CH(CH3)-。Furthermore, m = 0 to 3. Preferably, m = 0, 1, 2. For example, when m = 0, X 2 may be -C(O)-CH(CH 3 )-.

进一步，所述唑类化合物是具有式II或式III所示分子结构的化合物：
Furthermore, the azole compound is a compound having a molecular structure shown in Formula II or Formula III:

进一步，唑环连接的取代基和左侧苯环连接的取代基，在中间的苯环上连接位置为邻位或对位。即，所述唑类化合物是具有式IV或式V所示分子结构的化合物：
Furthermore, the substituent connected to the azole ring and the substituent connected to the left benzene ring are connected at the ortho position or the para position on the middle benzene ring. That is, the azole compound is a compound having a molecular structure shown in Formula IV or Formula V:

进一步，其中，n＝0～3；Further, wherein n=0-3;

Y为C或N；Y is C or N;

R1至少一个，R1各自独立的选自氢、C1-C4烷基、环己基、卤取代C1-C3烷基、C2-C4烯基、卤取代C2-C4烯基、硝基、氰基、氨基、卤素、甲氨基、羟基、羧基、羟基取代C1-C3烷基、苯基、甲基苯基、苯甲基、C1-C3烷氧基、C2-C4烯基氧基、磺酸基、硝基苯基、甲酰基、乙酰基、苯甲酰基、苄氧羰基、苄氧基、苯氧基、甲酰氧基、乙酰氧基、苯甲酰氧基、巯基、甲基磺酰基、磺酸基、C1-C3烷基氨基、双C1-C3烷基取代氨基或叔丁氧羰基。There is at least one R1 , and each R1 is independently selected from hydrogen, C1 - C4 alkyl, cyclohexyl, halogen-substituted C1 - C3 alkyl, C2 - C4 alkenyl, halogen-substituted C2 - C4 alkenyl, nitro, cyano, amino, halogen, methylamino, hydroxy, carboxyl, hydroxy-substituted C1 - C3 alkyl, phenyl, methylphenyl, benzyl, C1 - C3 alkoxy, C2 - C4 alkenyloxy, sulfonic acid, nitrophenyl, formyl, acetyl, benzoyl, benzyloxycarbonyl, benzyloxy, phenoxy, formyloxy, acetoxy, benzoyloxy, mercapto, methylsulfonyl, sulfonic acid, C1 - C3 alkylamino, di- C1 - C3 alkyl substituted amino or tert-butoxycarbonyl.

a表示R1的个数，a＝0～4；b表示R2的个数，b＝0～4。a represents the number of R1 , a=0-4; b represents the number of R2 , b=0-4.

R2至少一个，R2各自独立的选自C1-C3烷基、卤素、氨基、甲氨基、二甲氨基、羟基、羧基、硝基、氰基或卤代烷基。There is at least one R 2 , and each R 2 is independently selected from C 1 -C 3 alkyl, halogen, amino, methylamino, dimethylamino, hydroxyl, carboxyl, nitro, cyano or halogenated alkyl.

进一步，所述R1各自独立的选自：氢、甲基、卤取代甲基、乙基、正丙基、异丙基、异丁基、烯丙基、丙炔基、乙烯基、仲丁基、环己基、苯甲基、丙烯基、叔丁基、异丙烯基、乙炔基、苯基、对甲苯基、间甲苯基、邻甲苯基、邻硝基苯基、甲酰基、乙酰基、苯甲酰基、羧基、苄氧羰基、氰基、氨基、甲氨基、乙基氨基、丙基氨基、二甲氨基、二乙氨基、硝基、羟基、甲氧基、乙氧基、苄氧基、苯氧基、甲酰氧基、巯基、氟基、氯基、溴基、碘基、叔丁氧羰基、氟甲基或氟乙基。Further, each of the R1s is independently selected from the group consisting of hydrogen, methyl, halogen-substituted methyl, ethyl, n-propyl, isopropyl, isobutyl, allyl, propynyl, vinyl, sec-butyl, cyclohexyl, benzyl, propenyl, tert-butyl, isopropenyl, ethynyl, phenyl, p-tolyl, m-tolyl, o-tolyl, o-nitrophenyl, formyl, acetyl, benzoyl, carboxyl, benzyloxycarbonyl, cyano, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, nitro, hydroxyl, methoxy, ethoxy, benzyloxy, phenoxy, formyloxy, mercapto, fluoro, chloro, bromo, iodo, tert-butoxycarbonyl, fluoromethyl or fluoroethyl.

其中，氟甲基包括全氟甲基，氟乙基包括全氟乙基。Among them, the fluoromethyl group includes perfluoromethyl group, and the fluoroethyl group includes perfluoroethyl group.

优选地，所述R1各自独立的选自：氢、羟基、氰基、甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、氟、氯、溴、三氟甲基、氨基或叔丁氧羰基等。Preferably, the R 1 is independently selected from the group consisting of hydrogen, hydroxyl, cyano, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, fluorine, chlorine, bromine, trifluoromethyl, amino or tert-butyloxycarbonyl.

进一步，所述R2各自独立的选自：C1-C3烷基、卤素、氨基、硝基、氰基、氟基、氯基、溴基、碘基、羟基或全氟C1-C3烷基。Furthermore, each of the R 2 groups is independently selected from the group consisting of C 1 -C 3 alkyl, halogen, amino, nitro, cyano, fluoro, chloro, bromo, iodo, hydroxyl or perfluoro C 1 -C 3 alkyl.

进一步，a表示R1的个数，a＝0～3。优选地，a＝0、1、2。Furthermore, a represents the number of R 1 , and a=0 to 3. Preferably, a=0, 1, 2.

进一步，b表示R2的个数，b＝0～3。优选地，b＝0、1、2。例如b＝0，在中间苯环上无取代。Furthermore, b represents the number of R 2 , and b = 0 to 3. Preferably, b = 0, 1, 2. For example, when b = 0, there is no substitution on the middle benzene ring.

进一步，n＝0、1、2。例如n＝1。Furthermore, n = 0, 1, 2. For example, n = 1.

进一步，Y为C。Further, Y is C.

进一步，Y为N。Further, Y is N.

对于上述式I、式II、式III、式IV或式V化合物的取代基，可以互相进行替换。The substituents of the compounds of the above formula I, formula II, formula III, formula IV or formula V may be replaced with each other.

进一步，所述唑类化合物是以下化合物中的一种：
Further, the azole compound is one of the following compounds:

本发明的再一目的是将上述唑类化合物在制备药物中的应用。Another object of the present invention is to use the above azole compounds in the preparation of medicines.

上述唑类化合物应用于制备药物，特别是应用于制备动物抗真菌药物，具有优秀的抗菌活性，能够有效治疗毛皮动物真菌性皮肤病，具有低毒高效的优势。The above-mentioned azole compounds are used in the preparation of drugs, especially in the preparation of animal antifungal drugs, have excellent antibacterial activity, can effectively treat fungal skin diseases of fur animals, and have the advantages of low toxicity and high efficiency.

与现有技术相比，本发明的有益效果：Compared with the prior art, the present invention has the following beneficial effects:

1、本发明唑类化合物可以有效的解决现有技术缺乏动物专用抗真菌化学药物的问题，本发明筛选得到的氮唑类化合物具有低毒高效特点，更适合作为动物专用抗真菌药物的活性成分使用。1. The azole compounds of the present invention can effectively solve the problem of the lack of antifungal chemical drugs for animals in the prior art. The nitrogen azole compounds screened by the present invention have the characteristics of low toxicity and high efficiency, and are more suitable for use as active ingredients of antifungal drugs for animals.

2、本发明唑类化合物经过动物实验，相比于临床一线用药氟康唑、伊曲康唑、酮康唑具有更加优秀的抑菌活性，对于毛皮动物真菌感染使用效果好，特别是对犬小孢子菌、须毛癣菌、石膏样小孢子菌及白色念珠菌的菌丝生长抑制作用显著。2. Animal experiments have shown that the azole compounds of the present invention have better antibacterial activity than the clinical first-line drugs fluconazole, itraconazole and ketoconazole, and have good effects on fungal infections in fur animals, especially the mycelial growth inhibition of Microsporum canis, Trichophyton mentagrophytes, Microsporum gypseum and Candida albicans.

3、本发明唑类化合物对正常细胞毒性较小，优于酮康唑。动物实验表明本发明唑类化合物选择性优秀，在有效抑菌的同时，具有更低的细胞毒性，应用动物皮肤病，不影响动物正常生长，对于集中养殖的养殖场经济效益显著。3. The azole compounds of the present invention have less toxicity to normal cells and are better than ketoconazole. Animal experiments show that the azole compounds of the present invention have excellent selectivity, and have lower cytotoxicity while effectively inhibiting bacteria. They are applied to animal skin diseases without affecting the normal growth of animals, and have significant economic benefits for concentrated breeding farms.

附图说明BRIEF DESCRIPTION OF THE DRAWINGS

图1为化合物4与1EQP的对接图。Figure 1 is the docking diagram of compound 4 and 1EQP.

图2为化合物4与1CZ1的对接图。Figure 2 is the docking diagram of compound 4 and 1CZ1.

图3为化合物4与1AI9的对接图。FIG3 is a docking diagram of compound 4 and 1AI9.

图4为化合物4与4IIB的对接图。FIG4 is a docking diagram of compound 4 and 4IIB.

具体实施方式DETAILED DESCRIPTION

本发明的另一目的是提供一种上述唑类化合物的合成方法，包括以下步骤：取代苯基甲基氯或取代苯基甲基溴，和咪唑或三唑，在乙腈溶液中氢氧化钠催化作用下反应，得到唑类化合物。Another object of the present invention is to provide a method for synthesizing the above-mentioned azole compounds, comprising the following steps: substituted phenylmethyl chloride or substituted phenylmethyl bromide, and imidazole or triazole, reacting under the catalytic action of sodium hydroxide in an acetonitrile solution to obtain an azole compound.

其中，取代苯基甲基氯或取代苯基甲基溴的取代基是：苯甲基、苯基、叔丁基氧羰基苯基、苯甲氧基、Wherein, the substituents of substituted phenylmethyl chloride or substituted phenylmethyl bromide are: benzyl, phenyl, tert-butyloxycarbonylphenyl, benzyloxy,

本发明所述卤素是指氟、氯、溴、碘，所述卤取代是指卤素取代，具体的卤素是指氟取代、氯取代、溴取代、碘取代，可以是单取代也可以是多取代，可以是一种卤素取代也可以是混合多种卤素取代。The halogen mentioned in the present invention refers to fluorine, chlorine, bromine and iodine, and the halogen substitution refers to halogen substitution. Specifically, the halogen refers to fluorine substitution, chlorine substitution, bromine substitution and iodine substitution. It can be single substitution or multiple substitutions, and can be one halogen substitution or a mixture of multiple halogen substitutions.

所述C1-C3烷基是指：甲基、乙基、正丙基、异丙基。 The C 1 -C 3 alkyl group refers to: methyl, ethyl, n-propyl, isopropyl.

所述卤取代C1-C3烷基是指卤素取代的C1-C3烷基，如氟取代、氯取代、溴取代、碘取代。The halogen-substituted C 1 -C 3 alkyl group refers to a C 1 -C 3 alkyl group substituted with a halogen, such as a fluorine-substituted, chlorine-substituted, bromine-substituted, or iodine-substituted group.

所述C1-C3烷氧基是指：甲氧基、乙氧基、正丙基氧基、异丙基氧基。The C 1 -C 3 alkoxy group refers to: methoxy group, ethoxy group, n-propyloxy group, isopropyloxy group.

所述C2-C4烯基是指：乙烯基、丙烯基、丁烯基、1-丁烯基、2-丁烯基。The C 2 -C 4 alkenyl group refers to: vinyl, propenyl, butenyl, 1-butenyl, 2-butenyl.

所述C2-C4烯基氧基是指：乙烯基氧基、丙烯基氧基、丁烯基氧基等。The C 2 -C 4 alkenyloxy group refers to vinyloxy, propenyloxy, butenyloxy and the like.

下面结合试验例及具体实施方式对本发明作进一步的详细描述。但不应将此理解为本发明上述主题的范围仅限于以下的实施例，凡基于本发明内容所实现的技术均属于本发明的范围。The present invention is further described in detail below in conjunction with test examples and specific implementation methods. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.

实施例1Example 1

咪唑类化合物1-4合成Synthesis of Imidazole Compounds 1-4

反应的基本过程如下所示。
The basic process of the reaction is shown below.

咪唑类化合物合成路线Synthesis route of imidazole compounds

(1)在微波反应管中加入1.2eq(反应当量，缩写自equivalent，下同)咪唑溶于适量乙腈中再加入1eq氢氧化钠活化0.5h，再向反应管中加入1eq 1-苄基-2-氯甲基苯，在室温下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基咪唑衍生物(1)，即化合物1，化合物1结构经1H NMR，13C NMR及HR-MS确定。(1) 1.2 eq (reaction equivalent, abbreviated as equivalent, the same below) of imidazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile, and then 1 eq of sodium hydroxide was added to activate for 0.5 h. Then 1 eq of 1-benzyl-2-chloromethylbenzene was added to the reaction tube, reacted at room temperature for 5 h, diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyl imidazole derivative (1), i.e., compound 1. The structure of compound 1 was confirmed by 1 H NMR, 13 C NMR and HR-MS.

(2)在微波反应管中加入1.2eq咪唑溶于适量乙腈中再加入1eq氢氧化钠活化0.5h，再向反应管中加入1eq 2-溴甲基-1,1'-联苯，在室温下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基咪唑衍生物(2)，即化合物2，化合物2结构经1HNMR，13C NMR及HR-MS确定。(2) 1.2 eq of imidazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. Then 1 eq of sodium hydroxide was added to activate for 0.5 h. Then 1 eq of 2-bromomethyl-1,1'-biphenyl was added to the reaction tube and reacted at room temperature for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyl imidazole derivative (2), namely compound 2. The structure of compound 2 was confirmed by 1 HNMR, 13 C NMR and HR-MS.

(3)在微波反应管中加入1.2eq咪唑溶于适量乙腈中再加入1eq氢氧化钠活化0.5h，再向反应管中加入1eq 4'-溴甲基-[1,1'-联苯]-2-羧酸叔丁酯，在室温下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基咪唑衍生物(3)，即化合物3，化合物结构经1H NMR，13C NMR及HR-MS确定。 (3) 1.2 eq of imidazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. 1 eq of sodium hydroxide was added to activate for 0.5 h. 1 eq of 4'-bromomethyl-[1,1'-biphenyl]-2-carboxylic acid tert-butyl ester was added to the reaction tube and reacted at room temperature for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyl imidazole derivative (3), i.e., compound 3. The structure of the compound was confirmed by 1 H NMR, 13 C NMR and HR-MS.

(4)在微波反应管中加入1.2eq咪唑溶于适量乙腈中再加入1eq氢氧化钠活化0.5h，再向反应管中加入1eq 1-(4-苄氧基)苯基-2-溴-1-丙酮，在室温下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基咪唑衍生物(4)，即化合物4，化合物4结构经1H NMR，13C NMR及HR-MS确定。(4) 1.2 eq of imidazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. 1 eq of sodium hydroxide was added to activate for 0.5 h. 1 eq of 1-(4-benzyloxy)phenyl-2-bromo-1-propanone was added to the reaction tube and reacted at room temperature for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyl imidazole derivative (4), i.e., compound 4. The structure of compound 4 was confirmed by 1 H NMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物1-4结果如下： 1 H NMR, 13 C NMR and HR-MS, the test results of compounds 1-4 are as follows:

化合物1，HR-MS m/z:calculated for C17H16N2Na 271.1211,found 271.1206[M+Na]+；1H NMR(600MHz,Chloroform-d)δ7.58(d,J＝1.2Hz,1H),7.55–7.40(m,6H),7.30–7.24(m,3H),7.19–7.15(m,1H),6.95(t,J＝1.3Hz,1H),5.21(s,2H),4.17(s,2H).13C NMR(151MHz,Chloroform-d)δ139.36,138.24,137.38,134.24,131.09,129.53,128.64,128.58,128.52,128.42,127.25,126.42,119.21,48.31,38.73.Compound 1, HR-MS m/z: calculated for C 17 H 16 N 2 Na 271.1211, found 271.1206[M+Na] + ; 1 H NMR (600MHz, Chloroform-d) δ7.58 (d, J＝1.2Hz , 1 H),7.55–7.40(m,6H),7.30–7.24(m,3H),7.19–7.15(m, 1 H),6.95(t,J＝1.3Hz, 1 H),5.21(s,2H) ,4.17(s,2H). 13 C NMR(151MHz,Chloroform-d)δ139.36,138.24,137.38,134.24,131.09,129. 53,128.64,128.58,128.52,128.42,127.25,126.42,119.21,48.31,38.73.

化合物2，HR-MS m/z:calculated for C16H14N2Na 257.1055,found 257.1059[M+Na]+；1H NMR(600MHz,Chloroform-d)δ7.50–7.38(m,5H),7.35(dd,J＝7.3,1.7Hz,1H),7.29(s,1H),7.27–7.23(m,2H),7.20(dd,J＝7.6,1.5Hz,1H),7.05(d,J＝1.1Hz,1H),6.75(d,J＝1.3Hz,1H),5.09(s,2H).13C NMR(151MHz,Chloroform-d)δ141.81,139.89,137.15,133.26,130.33,129.25,128.77,128.45,128.38,128.19,127.91,127.50,118.92,48.62.Compound 2, HR-MS m/z:calculated for C 16 H 14 N 2 Na 257.1055, found 257.1059[M+Na] + ; 1 H NMR (600MHz, Chloroform-d) δ7.50–7.38 (m,5H) ,7.35(dd,J＝7.3,1.7Hz, 1 H),7.29(s, 1 H),7.27–7.23(m,2H),7.20(dd,J＝7.6,1.5Hz, 1H ),7.05(d,J＝1.1Hz, 1H ),6.75( d,J＝1.3Hz, 1H ),5.09(s,2H). 13C NMR(151MHz,Chloroform-d)δ141.81,139.89,137.15,133.26,130.33,129.25,128.77,128.45,128.38,128.19,127.91,127.50,118.92,48.62.

化合物3，HR-MS m/z:calculated for C2 1H22N2NaO2357.1579,found 357.1579[M+Na]+；1HNMR(600MHz,Chloroform-d)δ7.78(dd,J＝7.7,1.4Hz,1H),7.58(d,J＝1.2Hz,1H),7.48(td,J＝7.6,1.5Hz,1H),7.39(td,J＝7.6,1.3Hz,1H),7.31–7.26(m,2H),7.18(m,2H),7.08(m,2H),6.94(d,J＝1.3Hz,1H),5.16(s,2H),1.24(s,9H).13C NMR(151MHz,Chloroform-d)δ168.08,142.37,141.56,137.66,135.26,133.08,131.13,130.78,130.09,130.07,129.55,127.73,127.28,119.59,81.70,50.96,27.95.Compound 3, HR-MS m/z: calculated for C 2 1 H 22 N 2 NaO 2 357.1579, found 357.1579[M+Na] + ; 1 HNMR (600MHz, Chloroform-d) δ7.78 (dd, J=7.7 ,1.4Hz, 1 H),7.58(d,J＝1.2Hz, 1 H),7.48(td,J＝7.6,1.5Hz, 1H ),7.39(td,J＝7.6,1.3Hz, 1H ),7.31–7.26(m,2H),7.18(m,2H),7.08 (m,2H),6.94(d,J=1.3Hz, 1 H),5.16(s,2H),1.24(s,9H). 13 C NMR(151MHz,Chloroform-d)δ168.08,142.37,141.56,137.66,135.26,133.08,131 .13,130.78,130.09,130.07,129.55,127.73,127.28,119.59,81.70,50.96,27.95.

化合物4，HR-MS m/z:calculated for C19H18N2NaO2329.1267,found 329.1266[M+Na]+；1H NMR(600MHz,Chloroform-d)δ7.94–7.89(m,2H),7.63(s,1H),7.44–7.37(m,4H),7.35(ddt,J＝8.5,5.6,2.1Hz,1H),7.08(s,1H),7.05–6.99(m,3H),5.76(q,J＝7.1Hz,1H),5.13(s,2H),1.75(d,J＝7.1Hz,3H).13C NMR(151MHz,Chloroform-d)δ193.56,193.52,163.51,163.46,136.49,136.45,135.93,135.89,131.03,130.99,129.61,129.57,128.85,128.81,128.48,128.44,127.57,127.53,127.24,127.21,118.30,118.26,115.21,115.18,77.34,77.12,76.91,70.37,70.34,55.88,55.84,19.34,19.30.Compound 4, HR-MS m/z: calculated for C 19 H 18 N 2 NaO 2 329.1267, found 329.1266[M+Na] + ; 1 H NMR (600MHz, Chloroform-d) δ7.94–7.89 (m, 2H ),7.63(s, 1 H),7.44–7.37(m,4H),7.35(ddt,J＝8.5,5.6,2.1Hz, 1 H),7.08(s, 1 H),7.05–6.99(m,3H),5.76(q, J＝7.1Hz, 1 H),5.13(s,2H),1.75(d,J=7.1Hz,3H). 13 C NMR(151MHz,Chloroform-d)δ193.56,193.52,163.51,163.46,136.49,136.45,135.93,135.89,131.03,130.99,129.61,129.57,128.85,128.81, 128.48,128.44,127.57,127.53,127.24,127.21,118.30,118.26,115.21 ,115.18,77.34,77.12,76.91,70.37,70.34,55.88,55.84,19.34,19.30.

实施例2Example 2

1,2,4-三氮唑衍生物的合成(化合物5-8)Synthesis of 1,2,4-triazole derivatives (Compound 5-8)

基本反应的基本过程如下所示。
The basic process of the elementary reaction is shown below.

三唑类化合物合成路线Synthesis route of triazole compounds

在微波反应管中加入1.2eq1,2,4-三氮唑溶于适量乙腈中再加入1eq氢化钠活化0.5h，再向反应管中加入1eq 1-苄基-2-氯甲基苯，在50℃下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基三唑衍生物(5)，即化合物5，化合物5结构经1HNMR，13C NMR及HR-MS确定。1.2 eq 1,2,4-triazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. Then 1 eq sodium hydride was added to activate for 0.5 h. Then 1 eq 1-benzyl-2-chloromethylbenzene was added to the reaction tube. The mixture was reacted at 50° C. for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyltriazole derivative (5), i.e., compound 5. The structure of compound 5 was confirmed by 1 HNMR, 13 C NMR and HR-MS.

在微波反应管中加入1.2eq1,2,4-三氮唑溶于适量乙腈中再加入1eq氢化钠活化0.5h，再向反应管中加入1eq 2-溴甲基-1，1'-联苯，在50℃下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基三唑衍生物(6)，即化合物6，化合物6结构经1HNMR，13C NMR及HR-MS确定。1.2 eq 1,2,4-triazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. Then 1 eq sodium hydride was added to activate for 0.5 h. Then 1 eq 2-bromomethyl-1,1'-biphenyl was added to the reaction tube. The mixture was reacted at 50° C. for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyltriazole derivative (6), i.e., compound 6. The structure of compound 6 was confirmed by 1 HNMR, 13 C NMR and HR-MS.

在微波反应管中加入1.2eq1,2,4-三氮唑溶于适量乙腈中再加入1eq氢化钠活化0.5h，再向反应管中加入1eq 4'-溴甲基-[1,1'-联苯]-2-羧酸叔丁酯，在50℃下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基三唑衍生物(7)，即化合物7，化合物7结构经1H NMR，13C NMR及HR-MS确定。1.2 eq 1,2,4-triazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. Then 1 eq sodium hydride was added to activate for 0.5 h. Then 1 eq 4'-bromomethyl-[1,1'-biphenyl]-2-carboxylic acid tert-butyl ester was added to the reaction tube. The mixture was reacted at 50° C. for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyltriazole derivative (7), i.e., compound 7. The structure of compound 7 was confirmed by 1 H NMR, 13 C NMR and HR-MS.

在微波反应管中加入1.2eq1,2,4-三氮唑溶于适量乙腈中再加入1eq氢化钠活化0.5h，再向反应管中加入1eq 1-(4-苄氧基)苯基-2-溴-1-丙酮，在50℃下反应5h，用适量蒸馏水稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得苄基三唑衍生物(8)，即化合物8，化合物8结构经1H NMR，13C NMR及HR-MS确定。1.2 eq 1,2,4-triazole was added to a microwave reaction tube and dissolved in an appropriate amount of acetonitrile. Then 1 eq sodium hydride was added to activate for 0.5 h. Then 1 eq 1-(4-benzyloxy)phenyl-2-bromo-1-propanone was added to the reaction tube. The mixture was reacted at 50° C. for 5 h. The mixture was diluted with an appropriate amount of distilled water, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain a benzyltriazole derivative (8), i.e., compound 8. The structure of compound 8 was confirmed by 1 H NMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物5-8结果如下： 1 H NMR, 13 C NMR and HR-MS, the test results of compounds 5-8 are as follows:

化合物5，HR-MS m/z:calculated for C16H16N3250.1340,found 250.1344[M+H]+；1H NMR(600MHz,Chloroform-d)δ7.93(s,1H),7.68(s,1H),7.34(td,J＝7.5,1.4Hz,1H),7.31–7.18(m,5H),7.15(dd,J＝7.6,1.4Hz,1H),7.10–7.04(m,2H),5.26(s,2H),4.03(s,2H).13C NMR(151 MHz,Chloroform-d)δ172.45,145.03,125.82,116.51,112.21,109.71,109.39,108.42,106.74,96.37,51.33,42.15.Compound 5, HR-MS m/z: calculated for C 16 H 16 N 3 250.1340, found 250.1344[M+H] + ; 1 H NMR (600MHz, Chloroform-d) δ7.93 (s, 1 H), 7.68 (s, 1 H),7.34(td,J＝7.5,1.4Hz, 1 H),7.31–7.18(m,5H),7.15(dd,J=7.6,1.4Hz, 1 H),7.10–7.04(m,2H),5.26(s,2H),4.03(s,2H). 13C NMR(151 MHz,Chloroform-d)δ172.45,145.03,125.82,116.51,112.21,109.71,109.39,108.42,106.74,96.37,51.33,42.15.

化合物6，HR-MS m/z:calculated for C15H13N3Na 258.0990,found 258.1007[M+Na]+；1H NMR(600MHz,Chloroform-d)δ7.89(s,1H),7.61(s,1H),7.49–7.34(m,5H),7.34–7.28(m,2H),7.25–7.19(m,2H),5.29(s,2H).13C NMR(151MHz,Chloroform-d)δ151.89,143.01,142.04,139.85,131.83,130.42,129.34,128.83,128.64,128.52,128.04,127.66,51.33.Compound 6, HR-MS m/z: calculated for C 15 H 13 N 3 Na 258.0990, found 258.1007[M+Na] + ; 1 H NMR (600MHz, Chloroform-d) δ7.89 (s, 1 H), 7.61(s, 1 H),7.49–7.34(m,5H),7.34–7.28(m,2H),7.25–7.19(m,2H),5.29(s,2H). 13 C NMR(151MHz,Chloroform-d)δ151.89,143.01,142.04,139.85,131.83,130.42,129.34,128.83,128.64,128.52,128.04,127.66,51.33.

化合物7，HR-MS m/z:calculated for C20H21N3NaO2358.1540,found 358.1531[M+Na]+；1H NMR(600MHz,Chloroform-d)δ8.17(s,1H),8.13(s,1H),7.99(s,1H),7.78(dd,J＝7.7,1.4Hz,1H),7.48(td,J＝7.5,1.4Hz,1H),7.39(td,J＝7.6,1.3Hz,1H),7.35–7.27(m,4H),5.39(s,2H),1.23(s,9H).13C NMR(151MHz,Chloroform-d)δ167.88,152.11,143.07,142.55,141.30,133.43,132.80,130.96,130.56,129.89,129.41,127.82,127.56,81.57,53.55,29.78,27.71.Compound 7, HR-MS m/z: calculated for C 20 H 21 N 3 NaO 2 358.1540, found 358.1531[M+Na] + ; 1 H NMR (600MHz, Chloroform-d) δ8.17 (s, 1 H) ,8.13(s, 1 H),7.99(s, 1 H),7.78(dd,J＝7.7,1.4Hz, 1 H),7.48(td,J＝7.5,1.4Hz, 1 H),7.39(td,J＝7.6,1.3Hz, 1 H),7.35– 7.27(m,4H),5.39(s,2H),1.23(s,9H). 13 C NMR(151MHz,Chloroform-d)δ167.88,152.11,143.07,142.55,141.30,133.43,132 .80,130.96,130.56,129.89,129.41,127.82,127.56,81.57,53.55,29.78,27.71.

化合物8，HR-MS m/z:calculated for C18H17N3NaO2330.1218,found 330.1215[M+Na]+；1H NMR(600MHz,Chloroform-d)δ8.35(s,1H),8.21(s,1H),8.07–7.88(m,3H),7.49–7.32(m,4H),7.17–6.92(m,2H),6.14(q,J＝7.3Hz,1H),5.15(s,2H),1.83(d,J＝7.3Hz,3H).13C NMR(151MHz,Chloroform-d)δ193.14,163.95,151.68,142.95,136.15,131.54,129.11,128.74,127.83,127.21,115.51,70.65,59.50,18.89.Compound 8, HR-MS m/z: calculated for C 18 H 17 N 3 NaO 2 330.1218, found 330.1215[M+Na] + ; 1 H NMR (600MHz, Chloroform-d) δ8.35 (s, 1 H) ,8.21(s, 1 H),8.07–7.88(m,3H),7.49–7.32(m,4H),7.17–6.92(m,2H),6.14(q,J＝7.3Hz, 1H ),5.15(s,2H), 1.83(d,J=7.3Hz,3H). 13 C NMR(151MHz,Chloroform-d)δ193.14,163.95,151.68,142.95,136.15,131.54,129.11,128.74,127.83,127.21,115.51,70.65,59.50,18.89.

实施例3Example 3

化合物4衍生物的制备Preparation of derivatives of compound 4

1.化合物4的氟取代衍生物1. Fluorine-substituted derivatives of compound 4

反应的基本过程如下所示，
The basic process of the reaction is as follows:

氟取代衍生物Fluorine substituted derivatives

在微波反应管中加入1.2eq N-氟吡啶三氟甲磺酸盐溶于适量无水二氯甲烷中，再向反应管中加入1eq化合物4，在40℃下反应20h，用适量饱和氯化钠稀释，用乙酸乙酯萃取三次，水洗，干燥，粗产品柱层析分离，即可得化合物4的氟取代化合物9、10，即化合物9、10，结构经1HNMR，13C NMR及HR-MS确定。1.2 eq N-fluoropyridine trifluoromethanesulfonate was added to a microwave reaction tube and dissolved in an appropriate amount of anhydrous dichloromethane. Then 1 eq of compound 4 was added to the reaction tube, and the mixture was reacted at 40°C for 20 h. The mixture was diluted with an appropriate amount of saturated sodium chloride, extracted with ethyl acetate three times, washed with water, dried, and the crude product was separated by column chromatography to obtain fluorine-substituted compounds 9 and 10 of compound 4, i.e. compounds 9 and 10, whose structures were confirmed by 1 HNMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物9-10结果如下： 1 H NMR, 13 C NMR and HR-MS, the results of test compounds 9-10 are as follows:

化合物9，HR-MS m/z:calculated for C19H16F2N2NaO2365.1078,found 365.1083[M+Na]+；1H NMR(500MHz,Chloroform-d)δ8.01–7.73(m,2H),7.66(t,J＝1.6Hz,1H),7.42(dd,J＝4.0,1.6Hz,1H),7.34(dt,J＝7.7,1.0Hz,1H),7.19–7.02(m,3H),6.98–6.84(m,2H),5.74(q,J＝5.3Hz,1H),5.22(d,J＝1.1Hz,2H),1.60(s,3H)；13C NMR(125MHz,Chloroform-d)δ194.03,162.82,161.64,160.74,137.27,131.30,130.51,129.19,128.90,120.95,120.24,115.34,111.72,104.81,63.80,56.84,20.59.Compound 9, HR-MS m/z: calculated for C 19 H 16 F 2 N 2 NaO 2 365.1078, found 365.1083[M+Na] + ; 1 H NMR (500MHz, Chloroform-d) δ8.01–7.73(m ,2H),7.66(t,J＝1.6Hz, 1 H),7.42(dd,J＝4.0,1.6Hz, 1H ),7.34(dt,J＝7.7,1.0Hz, 1H ),7.19–7.02(m,3H),6.98–6.84(m,2H) ,5.74(q,J＝5.3Hz, 1 H),5.22(d,J＝1.1Hz,2H),1.60(s,3H); 13 C NMR(125MHz,Chloroform-d)δ194.03,162.82,161.64,160.74,137.27,131.30,130 .51,129.19,128.90,120.95,120.24,115.34,111.72,104.81,63.80,56.84,20.59.

化合物10，HR-MS m/z:calculated for C19H17FN2NaO2347.1172,found 347.1175[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.92–7.87(m,2H),7.66(t,J＝1.6Hz,1H),7.51–7.37(m,3H),7.09–7.01(m,4H),5.74(q,J＝5.3Hz,1H),5.07(t,J＝1.0Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,163.06,162.71,137.27,133.69,131.30,130.11,129.19,128.90,120.95,115.40,115.33,70.02,56.84,20.59.Compound 10, HR-MS m/z: calculated for C 19 H 17 FN 2 NaO 2 347.1172, found 347.1175[M+Na] + ; 1 H NMR (500MHz, Chloroform-d) δ7.92–7.87(m,2H ),7.66(t,J＝1.6Hz, 1 H),7.51–7.37(m,3H),7.09–7.01(m,4H),5.74(q,J＝5.3Hz, 1 H),5.07(t,J＝1.0Hz,2H),1.60(s, 3H).; 13C NMR(125MHz,Chloroform-d)δ194.03,163.06,162.71,137.27,133.69,131.30,130.11,129.19,128.90,120.95,115.40,115.33,70.02,56.84,20.59.

2.化合物4的氯取代衍生物2. Chloro-substituted derivatives of compound 4

反应的基本过程如下所示，
The basic process of the reaction is as follows:

氯取代衍生物Chlorine substituted derivatives

向装有搅拌子和冷凝管是烧瓶中加入次氯酸钙0.039mol、水100mL、乙酸10mL。然后将烧瓶浸入冰水中水浴(0℃)并搅拌至内容物至次氯酸钙全部溶解并获得浅黄色溶液。然后在三分钟内加入化合物4的丙酮溶液0.039mol，0℃下搅拌1h。然后将反应混合物用水稀释，瓶底有不溶的油状物，然后用乙醚萃取，用饱和碳酸氢钠洗，干燥。粗产品柱层析分离，即可得化合物4的氯取代化合物11、12，即化合物11、12，结构经1H NMR，13C NMR及HR-MS确定。Add 0.039 mol of calcium hypochlorite, 100 mL of water and 10 mL of acetic acid to a flask equipped with a stirrer and a condenser. Then immerse the flask in an ice-water bath (0°C) and stir until the contents are completely dissolved and the calcium hypochlorite is obtained to obtain a light yellow solution. Then add 0.039 mol of acetone solution of compound 4 within three minutes and stir at 0°C for 1 h. Then dilute the reaction mixture with water, there is an insoluble oil at the bottom of the flask, then extract with ether, wash with saturated sodium bicarbonate and dry. The crude product is separated by column chromatography to obtain chlorine-substituted compounds 11 and 12 of compound 4, namely compounds 11 and 12, whose structures are confirmed by 1 H NMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物11、12结果如下： 1 H NMR, 13 C NMR and HR-MS, the test results of compounds 11 and 12 are as follows:

化合物11，HR-MS m/z:calculated for C19H16Cl2N2NaO2397.0487,found 397.0483[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.96–7.86(m,2H),7.66(t,J＝1.6Hz,1H),7.48(dt,J＝8.3,1.1Hz,1H),7.45–7.37(m,2H),7.33(dd,J＝8.3,2.1Hz,1H),7.09–7.03(m,3H),5.74(q,J＝5.3Hz,1H),5.20(t,J＝0.8Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03, 162.82,137.27,135.03,133.92,131.33,131.30,131.21,129.19,128.90,128.05,127.41,120.95,115.33,68.73,56.84,20.59.Compound 11, HR-MS m/z: calculated for C 19 H 16 Cl 2 N 2 NaO 2 397.0487, found 397.0483[M+Na]+; 1 H NMR (500MHz, Chloroform-d) δ7.96–7.86 (m ,2H),7.66(t,J＝1.6Hz, 1 H),7.48(dt,J＝8.3,1.1Hz, 1H ),7.45–7.37(m,2H),7.33(dd,J＝8.3,2.1Hz, 1H ),7.09–7.03(m,3H) ,5.74(q,J＝5.3Hz, 1 H), 5.20 (t, J = 0.8Hz, 2H), 1.60 (s, 3H).; 13 C NMR(125MHz,Chloroform-d)δ194.03, 162.82,137.27,135.03,133.92,131.33,131.30,131.21,129.19,128.90,128.05,127.41,120.95,115.33,68.73,56.84,20.59.

化合物12，HR-MS m/z:calculated for C19H17ClN2NaO2363.0876,found 363.0874[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.95–7.85(m,2H),7.66(t,J＝1.6Hz,1H),7.47–7.32(m,5H),7.11–6.93(m,3H),5.75–5.70(m,1H),5.07(t,J＝1.0Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.27,136.11,134.36,131.30,129.87,129.19,128.90,128.64,120.95,115.33,70.02,56.84,20.59.Compound 12, HR-MS m/z: calculated for C 19 H 17 ClN 2 NaO 2 363.0876, found 363.0874[M+Na]+; 1 H NMR (500MHz, Chloroform-d) δ7.95–7.85 (m, 2H ),7.66(t,J＝1.6Hz, 1 H),7.47–7.32(m,5H),7.11–6.93(m,3H),5.75–5.70(m, 1 H),5.07(t,J＝1.0Hz,2H),1.60(s,3H). ; 13C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.27,136.11,134.36,131.30,129.87,129.19,128.90,128.64,120.95,115.33,70.02,56.84,20.59.

3.化合物4的溴取代3. Bromine substitution of compound 4

反应的基本过程如下所示
The basic process of the reaction is as follows

溴取代衍生物Bromine substituted derivatives

将溴代丁酰亚胺(NBS)和无水FeCl3(摩尔比：1：0.1)放入装有搅拌子的烧瓶中，以摩尔比(NBS：化合物4＝3：1)加入化合物4，再加入反应体积20％的乙腈，加热至90℃，反应7h，反应完成后用乙醚萃取，无水硫酸钠干燥。粗产品柱层析分离，即可得化合物4的溴取代化合物13、14，即化合物13、14，结构经1HNMR，13C NMR及HR-MS确定。Bromobutyrimide (NBS) and anhydrous FeCl 3 (molar ratio: 1:0.1) were placed in a flask equipped with a stirrer, compound 4 was added at a molar ratio (NBS: compound 4 = 3:1), and then acetonitrile was added in an amount of 20% of the reaction volume, heated to 90°C, reacted for 7 hours, extracted with ether after the reaction was completed, and dried over anhydrous sodium sulfate. The crude product was separated by column chromatography to obtain bromine-substituted compounds 13 and 14 of compound 4, namely compounds 13 and 14, whose structures were confirmed by 1 HNMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物13、14结果如下：The results of 1 H NMR, 13 C NMR and HR-MS for test compounds 13 and 14 are as follows:

化合物13，HR-MS m/z:calculated for C19H16Br2N2NaO2484.9476,found 484.9491[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.95–7.88(m,2H),7.66(t,J＝1.6Hz,1H),7.59(d,J＝1.9Hz,1H),7.48(dd,J＝8.2,1.8Hz,1H),7.45–7.37(m,2H),7.13–6.88(m,3H),5.74(q,J＝5.3Hz,1H),5.23(d,J＝1.0Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,162.82,137.27,134.46,132.34,131.30,131.03,130.91,129.19,128.90,124.97,122.15,120.95,115.33,71.39,56.84,20.59.Compound 13, HR-MS m/z: calculated for C 19 H 16 Br 2 N 2 NaO 2 484.9476, found 484.9491[M+Na]+; 1 H NMR (500MHz, Chloroform-d) δ7.95–7.88(m ,2H),7.66(t,J＝1.6Hz, 1 H),7.59(d,J＝1.9Hz, 1H ),7.48(dd,J＝8.2,1.8Hz, 1H ),7.45–7.37(m,2H),7.13–6.88(m,3H),5.74 (q,J＝5.3Hz, 1 H), 5.23 (d, J = 1.0Hz, 2H), 1.60 (s, 3H).; 13 C NMR(125MHz,Chloroform-d)δ194.03,162.82,137.27,134.46,132.34,131.30,131 .03,130.91,129.19,128.90,124.97,122.15,120.95,115.33,71.39,56.84,20.59.

化合物14，HR-MS m/z:calculated for C19H17BrN2NaO2407.0371,found 407.0365[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.94–7.88(m,2H),7.66(t,J＝1.6Hz,1H),7.57–7.47(m,2H),7.47–7.35(m,3H),7.12–7.02(m,3H),5.74(q,J＝5.3Hz,1H),5.07(t,J＝1.0Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.27,136.68,131.92,131.30,130.14,129.19,128.90,123.33,120.95,115.33,70.02,56.84,20.59. Compound 14, HR-MS m/z: calculated for C 19 H 17 BrN 2 NaO 2 407.0371, found 407.0365[M+Na]+; 1 H NMR (500MHz, Chloroform-d) δ7.94–7.88 (m, 2H ),7.66(t,J＝1.6Hz, 1 H),7.57–7.47(m,2H),7.47–7.35(m,3H),7.12–7.02(m,3H),5.74(q,J＝5.3Hz, 1 H),5.07(t,J＝1.0 Hz,2H),1.60(s,3H).; 13 C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.27,136.68,131.92,131.30,130.14,129.19,128.90,123.33,120.95,115.33,70.02,56.84,20.59.

4.化合物4的氨基取代4. Amino Substitution of Compound 4

反应的基本过程如下所示，
The basic process of the reaction is as follows:

氨基取代衍生物Amino substituted derivatives

将化合物4(0.20mmol)、三甲基硅叠氮化物(TMSN3，0.24mmol)、CHCl3(500μL)和三氟甲磺酸(TfOH，1.8mmol)加到带有搅拌子的反应管中，然后用密封小瓶，然后在60℃下反应20min。反应后，将小瓶在冰浴中冷却，在搅拌下缓慢加入500μL甲醇。用适量饱和碳酸氢钠溶液洗，然后用二氯甲烷萃取，无水硫酸钠干燥。粗产品柱层析分离，即可得化合物4的氨基取代化合物15，即化合物15，结构经1HNMR，13C NMR及HR-MS确定。Compound 4 (0.20 mmol), trimethylsilyl azide (TMSN3, 0.24 mmol), CHCl 3 (500 μL) and trifluoromethanesulfonic acid (TfOH, 1.8 mmol) were added to a reaction tube with a stirrer, and then sealed in a vial, and then reacted at 60°C for 20 min. After the reaction, the vial was cooled in an ice bath, and 500 μL of methanol was slowly added under stirring. Wash with an appropriate amount of saturated sodium bicarbonate solution, then extract with dichloromethane, and dry with anhydrous sodium sulfate. The crude product was separated by column chromatography to obtain the amino-substituted compound 15 of compound 4, i.e., compound 15, whose structure was confirmed by 1 HNMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物15结果如下：The results of 1 H NMR, 13 C NMR and HR-MS for testing compound 15 are as follows:

化合物15，HR-MS m/z:calculated for C19H19BrN3NaO2344.1375,found 344.1359[M+Na]+；1H NMR(500MHz,Chloroform-d)δ8.02–7.83(m,2H),7.66(t,J＝1.6Hz,1H),7.42(dd,J＝4.0,1.6Hz,1H),7.36–7.26(m,2H),7.14–6.98(m,3H),6.61–6.48(m,2H),5.74(q,J＝5.3Hz,1H),5.07(t,J＝1.0Hz,2H),4.17(d,J＝5.7Hz,1H),4.07(d,J＝5.7Hz,1H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,163.06,147.95,137.27,131.30,129.92,129.19,128.90,128.10,120.95,115.61,115.33,70.02,56.84,20.59.Compound 15, HR-MS m/z: calculated for C 19 H 19 BrN 3 NaO 2 344.1375, found 344.1359[M+Na]+; 1 H NMR (500MHz, Chloroform-d) δ8.02–7.83 (m, 2H ),7.66(t,J＝1.6Hz, 1 H),7.42(dd,J＝4.0,1.6Hz, 1 H),7.36–7.26(m,2H),7.14–6.98(m,3H),6.61–6.48(m,2H),5.74(q,J =5.3Hz, 1 H),5.07(t,J＝1.0Hz,2H),4.17(d,J＝5.7Hz, 1H ),4.07(d,J＝5.7Hz, 1 H),1.60(s,3H).; 13 C NMR(125MHz,Chloroform-d)δ194.03,163.06,147.95,137.27,131.30,129.92,129.19,128.90,128.10,120.95,115.61,115.33,70.02,56.84,20.59.

5.化合物4的三氟甲基取代5. Trifluoromethyl Substitution of Compound 4

三氟甲基取代衍生物合成反应的基本过程如下所示，
The basic process of the synthesis of trifluoromethyl substituted derivatives is as follows:

将化合物4(100μL)，二甲基亚砜(2.0ml)，1N二甲基亚砜硫酸溶液(2.0ml)，3.0mol/l三氟甲基碘二甲基亚砜溶液(1.0ml)，30％过氧化氢水溶液(0.2ml)和1.0mol/l硫酸亚铁(II)水溶液(0.3ml)装入双颈烧瓶中，在氩气环境下将混合物搅拌20min。在搅拌过程中，反应体系的温度在40℃至50℃。此后将所得溶液冷却至室温。用适量饱和氯化钠稀释，用乙酸 乙酯萃取三次，水洗三次，干燥，粗产品柱层析分离，即可得化合物4的三氟甲基取代化合物16、17，即化合物16、17，结构经1HNMR，13C NMR及HR-MS确定。Compound 4 (100 μL), dimethyl sulfoxide (2.0 ml), 1N dimethyl sulfoxide sulfuric acid solution (2.0 ml), 3.0 mol/l trifluoromethyl iodide dimethyl sulfoxide solution (1.0 ml), 30% hydrogen peroxide aqueous solution (0.2 ml) and 1.0 mol/l ferrous sulfate (II) aqueous solution (0.3 ml) were placed in a double-necked flask, and the mixture was stirred for 20 min under an argon atmosphere. During the stirring process, the temperature of the reaction system was between 40°C and 50°C. Thereafter, the resulting solution was cooled to room temperature. Diluted with an appropriate amount of saturated sodium chloride, acetic acid was added. The product was extracted with ethyl ester three times, washed with water three times, and dried. The crude product was separated by column chromatography to obtain trifluoromethyl substituted compounds 16 and 17 of compound 4, namely compounds 16 and 17, whose structures were confirmed by 1 HNMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物16、17结果如下：The results of 1 H NMR, 13 C NMR and HR-MS for test compounds 16 and 17 are as follows:

化合物16，HR-MS m/z:calculated for C20H17F3N2NaO2397.1140,found 397.1144[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.96–7.85(m,2H),7.72–7.58(m,3H),7.53–7.35(m,3H),7.12–6.98(m,3H),5.74(q,J＝5.3Hz,1H),5.07(t,J＝1.0Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.27,137.10,131.30,130.64,129.19,128.90,128.86,126.17,124.13,120.95,115.33,70.12,56.84,20.59.Compound 16, HR-MS m/z: calculated for C 20 H 17 F 3 N 2 NaO 2 397.1140, found 397.1144[M+Na]+; 1 H NMR(500MHz,Chloroform-d)δ7.96–7.85(m,2H),7.72–7.58(m,3H),7.53–7.35(m,3H),7.12–6.98(m,3H),5.74(q, J＝5.3Hz, 1 H), 5.07 (t, J = 1.0Hz, 2H), 1.60 (s, 3H).; 13 C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.27,137.10,131.30,130.64, 129.19,128.90,128.86,126.17,124.13,120.95,115.33,70.12,56.84,20.59.

化合物17，HR-MS m/z:calculated for C2 1H16F6N2NaO2465.1014,found 465.1017[M+Na]+；1H NMR(500MHz,Chloroform-d)δ7.94–7.86(m,3H),7.66(t,J＝1.6Hz,1H),7.51(dd,J＝6.4,1.8Hz,1H),7.44–7.35(m,2H),7.10–6.99(m,3H),5.74(q,J＝5.3Hz,1H),5.28(t,J＝0.9Hz,2H),1.60(s,3H).；13C NMR(125MHz,Chloroform-d)δ194.03,162.82,137.27,133.06,131.30,130.09,130.06,129.64,129.19,128.90,125.35,124.16,123.89,123.43,120.95,115.33,69.38,56.84,20.59.Compound 17, HR-MS m/z: calculated for C 2 1 H 16 F 6 N 2 NaO 2 465.1014, found 465.1017[M+Na]+; 1 H NMR (500MHz, Chloroform-d) δ7.94–7.86( m,3H),7.66(t,J＝1.6Hz, 1 H),7.51(dd,J＝6.4,1.8Hz, 1H ),7.44–7.35(m,2H),7.10–6.99(m,3H),5.74(q,J＝5.3Hz, 1H ),5.28 (t,J＝0.9Hz,2H),1.60(s,3H).; 13 C NMR(125MHz,Chloroform-d)δ194.03,162.82,137.27,133.06,131.30,130.09,130.06,129 .64,129.19,128.90,125.35,124.16,123.89,123.43,120.95,115.33,69.38,56.84,20.59.

6.化合物4的氰基取代6. Cyano substitution of compound 4

氰基取代衍生物合成反应的基本过程如下所示，
The basic process of the synthesis of cyano substituted derivatives is as follows:

将Pd(OAc)2(4.5mg，0.02mmol，10mol％)、3-(三氟甲基)喹啉(7.9mg，0.040mmol，20mol％)、N-乙酰甘氨酸(7.0mg，0.060mmol，30mol％)、AgF(101.5mg，0.80mmol,4eq)、芳烃(0.20mmol,1eq)、CuCN(35.8mg，0.40mmol，2eq)和HFIP(2mL)。将反应容器密封，混合物在室温下搅拌2分钟后，加热至90℃，并将反应混合物在该温度下以1000rpm搅拌18小时。将反应混合物冷却至室温，粗产品柱层析分离，即可得化合物4的氰基取代化合物18，即化合物18，结构经1H NMR，13C NMR及HR-MS确定。Pd(OAc) 2 (4.5 mg, 0.02 mmol, 10 mol%), 3-(trifluoromethyl)quinoline (7.9 mg, 0.040 mmol, 20 mol%), N-acetylglycine (7.0 mg, 0.060 mmol, 30 mol%), AgF (101.5 mg, 0.80 mmol, 4 eq), aromatic hydrocarbons (0.20 mmol, 1 eq), CuCN (35.8 mg, 0.40 mmol, 2 eq) and HFIP (2 mL) were added. The reaction vessel was sealed, the mixture was stirred at room temperature for 2 minutes, heated to 90° C., and the reaction mixture was stirred at 1000 rpm for 18 hours at the same temperature. The reaction mixture was cooled to room temperature, and the crude product was separated by column chromatography to obtain the cyano-substituted compound 18 of compound 4, i.e., compound 18, the structure of which was confirmed by 1 H NMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物18结果如下：The results of 1 H NMR, 13 C NMR and HR-MS for testing compound 18 are as follows:

化合物18，HR-MS m/z:calculated for C20H17N3NaO2354.1218,found 354.1222[M+Na]+； Compound 18, HR-MS m/z: calculated for C 20 H 17 N 3 NaO 2 354.1218, found 354.1222 [M+Na]+;

1H NMR(500MHz,Chloroform-d)δ7.96–7.85(m,2H),7.69–7.60(m,3H),7.54–7.45(m,2H),7.42(dd,J＝4.0,1.6Hz,1H),7.10–7.00(m,3H),5.74(q,J＝5.3Hz,1H),5.07(t,J＝1.0Hz,2H),1.60(s,3H).； 1 H NMR(500MHz,Chloroform-d)δ7.96–7.85(m,2H),7.69–7.60(m,3H),7.54–7.45(m,2H),7.42(dd,J＝4.0,1.6Hz, 1 H),7.10–7.00(m,3H),5.74(q,J＝5.3Hz, 1H ),5.07(t,J＝1.0Hz,2H),1.60(s,3H).;

13C NMR(125MHz,Chloroform-d)δ194.03,163.06,139.40,137.27,132.22,131.30,129.19,129.09,128.90,120.95,118.50,115.33,110.20,70.02,56.84,20.59. 13C NMR(125MHz,Chloroform-d)δ194.03,163.06,139.40,137.27,132.22,131.30, 129.19,129.09,128.90,120.95,118.50,115.33,110.20,70.02,56.84,20.59.

7.化合物4的羟基取代7. Hydroxyl Substitution of Compound 4

羟基取代衍生物反应的基本过程如下。
The basic process of the reaction of hydroxyl substituted derivatives is as follows.

将乙腈(5mL)中的苯(1mmol)和催化剂Cu-Au@g-C3N4(30mg)加入到25mL装有磁力搅拌子圆底烧瓶中。随后，将30％H2O2(1mmol)加入到反应混合物中，并使用20W家用冷LED灯照射，通过TLC监测反应的进程。反应完成后，从反应混合物中分离出催化剂，然后离心。用乙酸乙酯萃取，减压干燥，得到化合物4的羟基取代化合物19，即化合物19，结构经1H NMR，13C NMR及HR-MS确定。Benzene (1 mmol) and catalyst Cu-Au@gC 3 N 4 (30 mg) in acetonitrile (5 mL) were added to a 25 mL round-bottom flask equipped with a magnetic stirrer. Subsequently, 30% H 2 O 2 (1 mmol) was added to the reaction mixture, and a 20 W household cold LED lamp was used for irradiation, and the progress of the reaction was monitored by TLC. After the reaction was completed, the catalyst was separated from the reaction mixture and then centrifuged. It was extracted with ethyl acetate and dried under reduced pressure to obtain the hydroxyl-substituted compound 19 of compound 4, i.e., compound 19, whose structure was confirmed by 1 H NMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物19结果如下：The results of 1 H NMR, 13 C NMR and HR-MS for test compound 19 are as follows:

化合物19，HR-MS m/z:calculated for C19H18N2NaO3345.1215,found 345.1221[M+Na]+；Compound 19, HR-MS m/z: calculated for C 19 H 18 N 2 NaO 3 345.1215, found 345.1221 [M+Na]+;

1H NMR(500MHz,Chloroform-d)δ8.79(s,1H),7.96–7.83(m,2H),7.66(t,J＝1.6Hz,1H),7.42(dd,J＝4.0,1.6Hz,1H),7.36–7.22(m,2H),7.09–7.01(m,3H),6.82–6.60(m,2H),5.74(q,J＝5.3Hz,1H),5.07(t,J＝1.0Hz,2H),1.60(s,3H).； 1 H NMR (500MHz, Chloroform-d) δ8.79 (s, 1 H), 7.96–7.83 (m, 2H), 7.66 (t, J = 1.6 Hz, 1 H), 7.42 (dd, J = 4.0, 1.6Hz, 1 H),7.36–7.22(m,2H),7.09–7.01(m,3H),6.82–6.60(m,2H),5.74(q,J＝5.3Hz, 1 H),5.07(t,J＝1.0 Hz,2H),1.60(s,3H).;

13C NMR(125MHz,Chloroform-d)δ194.03,163.06,157.08,137.27,131.30,129.97,129.65,129.19,128.90,120.95,115.90,115.33,70.02,56.84,20.59. 13C NMR(125MHz,Chloroform-d)δ194.03,163.06,157.08,137.27,131.30,129.97,129.65,129.19,128.90,120.95,115.90,115.33,70.02,56.84,20.59.

8.化合物4的烷基取代8. Alkyl Substitution of Compound 4

烷基取代过程，反应的基本过程如下，
Alkyl substitution process, the basic process of the reaction is as follows,

在大气压下，在带有玻璃管(8mm内径)的连续流喷射鼓泡反应器中反应。将催化剂：H型丝光沸石(H-MOR，0.5g)从环境温度原位热处理至673K(10K/min)。然后将化合物4：甲醇(1:1)的混合物加入反应器(WHSV＝2.0HR-1)，反应3h，冷却至室温。得到化合物4的烷基取代化合物20，即化合物20，结构经1HNMR，13C NMR及HR-MS确定。The reaction was carried out in a continuous flow jet bubbling reactor with a glass tube (8 mm inner diameter) at atmospheric pressure. The catalyst: H-mordenite (H-MOR, 0.5 g) was in situ heat treated from ambient temperature to 673 K (10 K/min). Then a mixture of compound 4:methanol (1:1) was added to the reactor (WHSV=2.0 HR-1), reacted for 3 h, and cooled to room temperature. The alkyl substituted compound 20 of compound 4, i.e., compound 20, was obtained, and its structure was confirmed by 1 HNMR, 13 C NMR and HR-MS.

1H NMR，13C NMR及HR-MS，测试化合物20结果如下： 1 H NMR, 13 C NMR and HR-MS, the results of test compound 20 are as follows:

化合物20，HR-MS m/z:calculated for C20H20N2NaO23431422,found 3431427[M+Na]+；Compound 20, HR-MS m/z: calculated for C 20 H 20 N 2 NaO 2 3431422, found 3431427 [M + Na] +;

1H NMR(500MHz,Chloroform-d)δ7.96–7.89(m,2H),7.66(t,J＝1.6Hz,1H),7.42(dd,J＝4.0,1.7Hz,1H),7.34–7.24(m,2H),7.20–7.12(m,2H),7.12–6.98(m,3H),5.74(q,J＝5.3Hz,1H),5.06(t,J＝1.0Hz,2H),1.59(d,J＝5.3Hz,3H).； 1 H NMR (500MHz, Chloroform-d) δ7.96–7.89 (m, 2H), 7.66 (t, J＝1.6Hz, 1 H), 7.42 (dd, J＝4.0, 1.7Hz, 1 H),7.34–7.24(m,2H),7.20–7.12(m,2H),7.12–6.98(m,3H),5.74(q,J＝5.3Hz, 1 H),5.06(t,J＝1.0 Hz, 2H), 1.59 (d, J = 5.3Hz, 3H).;

13C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.88,137.27,135.07,131.30,129.23,129.19,128.90,128.51,120.95,115.33,70.02,56.84,21.04,20.59. 13C NMR(125MHz,Chloroform-d)δ194.03,163.06,137.88,137.27,135.07,131.30 ,129.23,129.19,128.90,128.51,120.95,115.33,70.02,56.84,21.04,20.59.

实施例4Example 4

合成化合物体外抗真菌活性测定In vitro antifungal activity assay of synthetic compounds

1.实验菌种1. Experimental strains

石膏样小孢子菌标准菌株(Microsporum gypseum，M873.579)、须毛癣菌标准菌株(Trichophyton mentagrophytes，M3.859)、犬小孢子菌标准菌株(Microsporum canis，ATCC8137)、白色念珠菌(Candida albicans(Robin)Berkhout,ATCC10231)。Standard strain of Microsporum gypseum (M873.579), standard strain of Trichophyton mentagrophytes (M3.859), standard strain of Microsporum canis (ATCC8137), and Candida albicans (Robin) Berkhout (ATCC10231).

2实验方法2 Experimental methods

2.1药物溶液配备2.1 Preparation of drug solution

将氟康唑、酮康唑、伊曲康唑以及合成的化合物于25℃下减压干燥12h，精密称定干燥后样品，加入适量20％DMSO溶解，定容，配成药物摩尔浓度分别为6μmol/mL、3μmol/mL、1.5μmol/mL、0.75μmol/mL、0.375μmol/mL的原液各4mL。用0.22μm的微孔滤膜过滤后，置4℃冰箱保存备用。Fluconazole, ketoconazole, itraconazole and the synthesized compound were dried under reduced pressure at 25°C for 12h, the dried samples were accurately weighed, and an appropriate amount of 20% DMSO was added to dissolve, and the volume was fixed to prepare 4mL of stock solutions with drug molar concentrations of 6μmol/mL, 3μmol/mL, 1.5μmol/mL, 0.75μmol/mL, and 0.375μmol/mL, respectively. After filtering with a 0.22μm microporous filter membrane, the samples were stored in a refrigerator at 4°C for later use.

2.2培养基制备2.2 Culture medium preparation

(1)马铃薯琼脂培养基(PDA)制备：取新鲜去皮马铃薯，切成小块，称量200g，放入1L蒸馏水中煮30min，用三层纱布过滤后，补足因蒸发而减少的蒸馏水至1L。在滤液中 加入20g的葡萄糖和20g的琼脂，加热，待琼脂完全融化后，趁热分装于250mL的锥形瓶中并塞上棉塞，用报纸包好，于121℃高压蒸汽灭菌锅高压灭菌30min。灭菌完成后，转移至无菌操作台，待培养基温度降至50℃左右。倒无菌平板晃匀，平放，制成PDA平板，凝固后4℃保存，备用。(1) Preparation of potato agar medium (PDA): Take fresh peeled potatoes, cut them into small pieces, weigh 200 g, put them into 1 L of distilled water and boil them for 30 min. After filtering with three layers of gauze, add the distilled water reduced by evaporation to 1 L. Add 20g of glucose and 20g of agar, heat, and when the agar is completely melted, divide it into 250mL conical bottles while hot and plug them with cotton plugs, wrap them with newspapers, and sterilize them in a high-pressure steam sterilizer at 121℃ for 30min. After sterilization, transfer it to a sterile operating table and wait until the culture medium temperature drops to about 50℃. Pour the sterile plate and shake it evenly, lay it flat, and make a PDA plate. After solidification, store it at 4℃ for later use.

(2)含药平板的制备：分别取上述制备的不同药液1mL以及上述冷却至50℃左右的PDA培养基9mL倒入灭菌的培养皿中，制成浓度为0.6μmol/mL、0.3μmol/mL、0.15μmol/mL、0.075μmol/mL、0.0375μmol/mL的含药平板。待培养基冷却至50℃，加入1mL 30％DMSO以及9mLPDA培养基制成该含药平板的对照平板，凝固后4℃保存，备用。(2) Preparation of drug-containing plates: Take 1 mL of the different drug solutions prepared above and 9 mL of the PDA medium cooled to about 50°C and pour them into sterilized culture dishes to prepare drug-containing plates with concentrations of 0.6 μmol/mL, 0.3 μmol/mL, 0.15 μmol/mL, 0.075 μmol/mL, and 0.0375 μmol/mL. After the culture medium is cooled to 50°C, add 1 mL 30% DMSO and 9 mL PDA medium to prepare a control plate of the drug-containing plate, and store it at 4°C after solidification for later use.

2.3菌悬液的制备2.3 Preparation of bacterial suspension

将试验所需的石膏样小孢子菌、须毛癣菌、犬小孢子菌、白色念珠菌从冰箱中取出，用无菌接种环分别将3种真菌接种到SDA平板上，27℃培养2周，活化真菌，使菌落均匀覆盖在培养基上。吸取灭菌的生理盐水2mL冲洗菌落表面的菌丝和孢子，将含有菌丝和孢子的冲洗液用血细胞计数板计数，将浓度调至为105～106CFU/mL，获得菌悬液，备用。Take out the required Microsporum gypseum, Trichophyton mentagrophytes, Microsporum canis, and Candida albicans from the refrigerator, use a sterile inoculation loop to inoculate the three fungi onto the SDA plate, and culture at 27°C for 2 weeks to activate the fungi so that the colonies are evenly covered on the culture medium. Pipette 2mL of sterile saline to rinse the hyphae and spores on the surface of the colony, count the washing liquid containing hyphae and spores with a hemocytometer, adjust the concentration to 105-106 CFU/mL, and obtain a bacterial suspension for use.

2.4菌丝生长速率和抑菌率的测定2.4 Determination of mycelial growth rate and antibacterial rate

将供试真菌接种在PDA平板上27℃培养5天备用。用已灭菌的打孔器在接种供试菌的PDA平板边缘切取直径为1cm的菌饼，将该菌饼接入含药培养基上，有菌丝的一面朝下，每个培养皿接种1个菌饼。同时设置空白对照、阴性对照、阳性对照(氟康唑、伊曲康唑、酮康唑)。做3次重复试验，置于27℃的恒温培养箱中培养。培养7d，用十字交叉法测量供试真菌在含药培养基上的菌落直径，与对照比较计算各药液处理对菌丝直线生长的抑制率。按下式求出菌丝生长速率和抑菌率：菌丝生长速率(cm/d)＝(测量直径平均值-1.0)/天数；抑菌率(％)＝(空白对照菌落直径-处理菌落直径)÷空白对照菌落直径×100。The test fungi were inoculated on PDA plates and cultured at 27°C for 5 days. A sterilized punch was used to cut a 1 cm diameter bacterial cake from the edge of the PDA plate inoculated with the test fungi, and the cake was inoculated on the drug-containing culture medium, with the mycelium side facing down, and one bacterial cake was inoculated in each culture dish. At the same time, a blank control, a negative control, and a positive control (fluconazole, itraconazole, ketoconazole) were set up. Three repeated tests were performed and cultured in a constant temperature incubator at 27°C. After 7 days of culture, the colony diameter of the test fungi on the drug-containing culture medium was measured by the cross method, and the inhibition rate of each drug solution treatment on the linear growth of mycelium was calculated by comparing with the control. The mycelium growth rate and the inhibition rate were calculated according to the following formula: Mycelium growth rate (cm/d) = (average measured diameter - 1.0) / number of days; Inhibition rate (%) = (blank control colony diameter - treated colony diameter) ÷ blank control colony diameter × 100.

2.5 MIC测定2.5 MIC determination

取无菌96孔板，首先每孔加入100μL的PDB培养基，然后在第1列、第2列每孔分别加入药溶液100μL，混匀；2～10列进行倍比稀释：从第二列取100μL液体至第3列，混匀；从第3列取100μL液体至第4列，依次类推，至第10列(浓度范围6000μmol/L-11.78μmol/L)。在第11列加入100μLDMSO，10、11列混匀后取出100μL液体，弃去；最后12列每孔加入菌悬液100μL。第1列为空白对照，第11列为阴性对照，第12列为生长对照。将该96孔板置于27℃恒温培养箱中培养48h，以溶液澄清透明的孔的最小药液浓度为MIC。每个设置三组重复。Take a sterile 96-well plate, first add 100 μL of PDB medium to each well, then add 100 μL of drug solution to each well in the 1st and 2nd columns respectively, and mix well; perform serial dilution in columns 2 to 10: take 100 μL of liquid from the 2nd column to the 3rd column, mix well; take 100 μL of liquid from the 3rd column to the 4th column, and so on, to the 10th column (concentration range 6000 μmol/L-11.78 μmol/L). Add 100 μL of DMSO to the 11th column, take out 100 μL of liquid after mixing columns 10 and 11, and discard; finally, add 100 μL of bacterial suspension to each well in the 12th column. The 1st column is a blank control, the 11th column is a negative control, and the 12th column is a growth control. The 96-well plate was placed in a 27°C constant temperature incubator for 48 hours, and the minimum drug concentration of the well with clear and transparent solution was the MIC. Three groups of replicates were set for each.

3.抑菌试验结果3. Antibacterial test results

3.1化合物1-8对菌丝生长速率的影响 3.1 Effects of compounds 1-8 on mycelial growth rate

菌丝生长速率法测试结果见表1-3。The test results of mycelium growth rate method are shown in Tables 1-3.

其中阳性对照组伊曲康唑和氟康唑对对犬小孢子菌、须毛癣菌、石膏样小孢子菌的菌丝生长抑制作用不明显，酮康唑有明显的抑制作用且效果较好，尤其是对须毛癣菌的菌丝生长速率的影响更甚。化合物1-8对这三种菌的菌丝生长有不同程度的影响，且均比伊曲康唑和氟康唑效好。在6μmol/mL时化合物5的对犬小孢子菌菌丝的生长抑制作用非常明显，化合物3、4对须毛癣菌和石膏样小孢子菌的菌丝生长速率的影响均最大，均小于1mm/d。Among them, itraconazole and fluconazole in the positive control group had no obvious inhibitory effect on the hyphal growth of Microsporum canis, Trichophyton mentagrophytes, and Microsporum gypseum, while ketoconazole had a significant inhibitory effect and a better effect, especially on the hyphal growth rate of Trichophyton mentagrophytes. Compounds 1-8 had different degrees of influence on the hyphal growth of these three fungi, and were better than itraconazole and fluconazole. At 6 μmol/mL, compound 5 had a very obvious inhibitory effect on the hyphal growth of Microsporum canis, and compounds 3 and 4 had the greatest influence on the hyphal growth rate of Trichophyton mentagrophytes and Microsporum gypseum, both less than 1 mm/d.

表1不同浓度的化合物对犬小孢子菌的菌丝生长速率的影响
Table 1 Effects of different concentrations of compounds on the mycelial growth rate of Microsporum canis

表2不同浓度的化合物对须毛癣菌的菌丝生长速率的影响
Table 2 Effects of different concentrations of compounds on the hyphae growth rate of Trichophyton mentagrophytes

表3不同浓度的化合物对石膏样小孢子菌的菌丝生长速率的影响

Table 3 Effects of different concentrations of compounds on the mycelial growth rate of Microsporum gypseum

4.1.2化合物1-8对三种菌的抑菌率4.1.2 Antibacterial rate of compounds 1-8 against three bacteria

抑菌率的测定结果见表4-6。The results of the inhibition rate determination are shown in Tables 4-6.

阳性对照组中酮康唑的抑菌率最大，抑菌效果均比伊曲康唑和氟康唑好。化合物1-8对三种真菌均具有不同程度的抑菌作用。在浓度为6μmol/mL时，化合物3、4对三种菌的抑菌率均大于60％，抑菌效果好，化合物2对犬小孢子菌、须毛癣菌的抑菌率大于60％抑菌效果好，但对于石膏样小孢子菌的抑菌率大于45％，抑菌效果较好。化合物1对须毛癣菌抑菌效果好，对于犬小孢子菌和石膏样小孢子菌的抑菌效果较好。在浓度为3μmol/mL时，化合物4对三种菌仍然有好的抑菌效果，化合物3对须毛癣菌和石膏样小孢子菌的抑菌效果好，但对于犬小孢子菌抑菌效果较差，化合物1均有较好的抑菌效果。In the positive control group, ketoconazole had the highest antibacterial rate, and its antibacterial effect was better than that of itraconazole and fluconazole. Compounds 1-8 had different degrees of antibacterial effect on the three fungi. At a concentration of 6 μmol/mL, the antibacterial rates of compounds 3 and 4 on the three fungi were greater than 60%, with good antibacterial effects. The antibacterial rate of compound 2 on Microsporum canis and Trichophyton mentagrophytes was greater than 60%, with good antibacterial effects, but the antibacterial rate for Microsporum gypseum was greater than 45%, with good antibacterial effects. Compound 1 had a good antibacterial effect on Trichophyton mentagrophytes, and had a good antibacterial effect on Microsporum canis and Microsporum gypseum. At a concentration of 3 μmol/mL, compound 4 still had a good antibacterial effect on the three fungi, compound 3 had a good antibacterial effect on Trichophyton mentagrophytes and Microsporum gypseum, but had a poor antibacterial effect on Microsporum canis, and compound 1 had a good antibacterial effect.

表4不同浓度的化合物对犬小孢子菌的抑菌率
Table 4 Inhibition rate of compounds at different concentrations against Microsporum canis

表5不同浓度的化合物对须毛癣菌的抑菌率

Table 5 Inhibition rate of compounds at different concentrations against Trichophyton mentagrophytes

表6不同浓度的化合物对石膏样小孢子菌的抑菌率
Table 6 Inhibition rate of compounds at different concentrations against Microsporum gypseum

4.1.3化合物1-20对四种真菌的MIC测定结果4.1.3 MIC results of compounds 1-20 against four fungi

MIC的测定结果见表7。化合1-20对四种真菌均具有不同程度的抑制作用。与阳性对照组(伊曲康唑、氟康唑及酮康唑)比较，化合物4及其衍生物化合物9,10,13,14,16,17对犬小孢子菌、须毛癣菌、石膏样小孢子菌及白色念珠菌的MIC均优于阳性对照；化合物11,12,18次之；化合物1-3,15,19,20均优于伊曲康唑及氟康唑。The results of MIC determination are shown in Table 7. Compounds 1-20 have different degrees of inhibitory effects on the four fungi. Compared with the positive control group (itraconazole, fluconazole and ketoconazole), the MICs of compound 4 and its derivatives compounds 9, 10, 13, 14, 16, 17 against Microsporum canis, Trichophyton mentagrophytes, Microsporum gypseum and Candida albicans are better than the positive control; compounds 11, 12, 18 are second; compounds 1-3, 15, 19, 20 are better than itraconazole and fluconazole.

表7化合物1-20对四种菌的MIC

Table 7 MIC of compounds 1-20 against four bacteria

通过抑菌试验可知，20个化合物中，除化合物5-8外，其余化合物对四种病原真菌均具有优于阳性对照或与阳性对照相当的抗真菌活性。The antifungal test showed that, among the 20 compounds, except for compounds 5-8, the other compounds had antifungal activities against the four pathogenic fungi that were superior to or equivalent to the positive control.

实施例5Example 5

将上述实施例合成的化合物1-20，进行正常细胞的细胞毒性测定The compounds 1-20 synthesized in the above examples were subjected to normal cell cytotoxicity assay.

1.细胞株1. Cell lines

NIH3T3细胞NIH3T3 cells

2.实验方法2. Experimental Methods

2.1接种细胞2.1 Cell inoculation

待NIH3T3细胞生长良好并处于对数生长期时，按照传代操作，将细胞经重悬于1mL的完全培养基中，加入适量完全培养液稀释，计数为5×104个/mL。取配制好的细胞悬液接种于96孔细胞培养板，每孔中加入100μL，封口后转移至37℃、5％CO2恒温培养箱中培养24h。When NIH3T3 cells grow well and are in the logarithmic growth phase, follow the subculture procedure, resuspend the cells in 1 mL of complete culture medium, add an appropriate amount of complete culture medium to dilute, and count to 5 × 10 4 cells/mL. Take the prepared cell suspension and inoculate it into a 96-well cell culture plate, add 100 μL to each well, seal it, and transfer it to a 37°C, 5% CO 2 constant temperature incubator for 24 hours.

2.2配置药液和给药2.2 Preparation of drug solution and administration

将化合物用DMSO溶解，并以相应溶剂作为阴性对照组。对照组加入100μL完全培养基，实验组补充完全培养基至200μL，使每孔药液的终浓度20μM，平行5组。在20μM浓度下显示出良好抑制活性的衍生物，按照一定的浓度梯度使用NIH3T3细胞对活性药物进行复筛。The compound was dissolved in DMSO, and the corresponding solvent was used as the negative control group. 100 μL of complete medium was added to the control group, and the complete medium was supplemented to 200 μL in the experimental group, so that the final concentration of the drug solution in each well was 20 μM, and 5 groups were paralleled. Derivatives that showed good inhibitory activity at a concentration of 20 μM were rescreened for active drugs using NIH3T3 cells according to a certain concentration gradient.

2.3 MTS试剂盒检测细胞活力2.3 MTS kit to detect cell viability

细胞经药物处理48h后，按1:10的比例将MTS溶液和完全培养液混合，弃去96孔板中含药液的培养基，每孔加入200μL混合液，并设置5个不含细胞的混合液空白组，置于恒温箱中反应2-4h。用酶标仪检测在490nm吸收波长下各孔的吸光度，导出数据并计算抑制率，通过GraphPad Prism 8软件利用Bliss法计算IC50。After the cells were treated with drugs for 48 hours, the MTS solution and complete culture medium were mixed at a ratio of 1:10, the medium containing the drug solution in the 96-well plate was discarded, 200 μL of the mixed solution was added to each well, and 5 blank groups of the mixed solution without cells were set up and placed in a constant temperature box for reaction for 2-4 hours. The absorbance of each well at an absorption wavelength of 490 nm was detected by an ELISA instrument, the data were exported and the inhibition rate was calculated, and the IC 50 was calculated by the Bliss method using GraphPad Prism 8 software.

3.实验结果3. Experimental results

化合物对NIH3T3细胞的IC50结果见表8。 The IC50 results of the compounds on NIH3T3 cells are shown in Table 8.

所测化合物中，化合物3的IC50最小，细胞毒性最大，其余化合物叫酮康唑均细胞毒性较小，可见这些化合物对正常组织的毒性应低于酮康唑。Among the tested compounds, compound 3 has the smallest IC 50 and the greatest cytotoxicity, while the remaining compounds, including ketoconazole, have lower cytotoxicity. This shows that the toxicity of these compounds to normal tissues should be lower than that of ketoconazole.

表8化合物1-20对NIH3T3细胞的IC50
Table 8 IC 50 of compounds 1-20 on NIH3T3 cells

结论in conclusion

综上可知，阳性对照氟康唑和伊曲康唑对于犬小孢子菌、须毛癣菌、石膏样小孢子菌的抑制作用不明显，酮康唑对这三种菌有明显抑制作用。三种阳性对照对白色念珠菌均有一定抑制作用。而合成的化合物1-20对犬小孢子菌、须毛癣菌、石膏样小孢子菌及白色念珠菌均具有不同程度的抑菌作用，其中化合物4,9,10,13,14,16,17对这四种菌均有明显的抑制作用，且均优于氟康唑、伊曲康唑及酮康唑(部分相当)；化合物11,12,18次之；化合物1-3,15,19,20再次之。且化合物1-20对正常细胞均表现出较低的细胞毒性，安全性较高。以上化合物均具有潜在的抗真菌应用价值。In summary, the positive controls fluconazole and itraconazole have no obvious inhibitory effect on Microsporum canis, Trichophyton mentagrophytes, and Microsporum gypseum, while ketoconazole has a significant inhibitory effect on these three bacteria. The three positive controls all have a certain inhibitory effect on Candida albicans. The synthesized compounds 1-20 have different degrees of antibacterial effects on Microsporum canis, Trichophyton mentagrophytes, Microsporum gypseum, and Candida albicans. Among them, compounds 4, 9, 10, 13, 14, 16, and 17 have obvious inhibitory effects on these four bacteria, and are better than fluconazole, itraconazole, and ketoconazole (partially equivalent); compounds 11, 12, and 18 are second; compounds 1-3, 15, 19, and 20 are third. And compounds 1-20 show low cytotoxicity to normal cells and have high safety. The above compounds all have potential antifungal application value.

实施例6Example 6

代表性化合物抗真菌作用靶点预测Prediction of antifungal targets of representative compounds

选择代表性化合物1-4，采用分子对接的方法，考察代表化合物与常见抗真菌作用靶点蛋白的结合情况和作用模式，预测本申请中活性化合物的可能作用靶点。Representative compounds 1-4 were selected, and the molecular docking method was used to examine the binding and action mode of the representative compounds with common antifungal target proteins, and to predict the possible targets of the active compounds in this application.

1.分子对接方法1. Molecular docking method

从RCSB数据库(http://www.rcsb.org/)分别找到：致病性酵母白色念珠菌的甾醇14-α脱甲基酶(CYP51，PDB ID:5FSA)；白色念珠菌的甾醇14-α脱甲基酶(CYP51，PDB ID:5TZ1)；白色念珠菌的外切-β(1,3)-葡聚糖酶(XOG1，PDB ID：1EQP)；白色念珠菌的 外切-β(1,3)-葡聚糖酶(XOG1，PDB ID:1CZ1)；来自白色念珠菌的几丁质合成酶2(CHS2，PDB ID：7STL)；白色念珠菌的二氢叶酸还原酶(DFR1，PDB ID：1AI9)；白色念珠菌N-肉豆蔻酰转移酶(NMT1，PDB ID：1IYK)；白色念珠菌的磷酸甘露糖异构酶(PMI，PDB ID：1PMI)；来自曲霉的β-(1,4)-半乳糖酶(GAL1，PDB ID：1FOB)；来自曲霉的β-葡萄糖苷酶1(BGL1，PDB ID：4IIB)；来自曲霉的鼠李半乳糖醛酸酶A(RHGA，PDB ID：1RMG)的晶体结构。From the RCSB database (http://www.rcsb.org/), we found the following: sterol 14-α demethylase (CYP51, PDB ID: 5FSA) of pathogenic yeast Candida albicans; sterol 14-α demethylase (CYP51, PDB ID: 5TZ1) of Candida albicans; exo-β(1,3)-glucanase (XOG1, PDB ID: 1EQP) of Candida albicans; Exo-β(1,3)-glucanase (XOG1, PDB ID: 1CZ1); chitin synthase 2 from Candida albicans (CHS2, PDB ID: 7STL); dihydrofolate reductase from Candida albicans (DFR1, PDB ID: 1AI9); Candida albicans N-myristoyltransferase (NMT1, PDB ID: 1IYK); phosphomannose isomerase from Candida albicans (PMI, PDB ID: 1PMI); β-(1,4)-galactosidase from Aspergillus (GAL1, PDB ID: 1FOB); β-glucosidase 1 from Aspergillus (BGL1, PDB ID: 4IIB); crystal structure of rhamnogalacturonase A from Aspergillus (RHGA, PDB ID: 1RMG).

从Pub Chem数据库(https://pubchem.ncbi.nlm.nih.gov/)中下载化合物1-4以及阳性对照氟康唑、酮康唑、伊曲康唑结构的SDF格式文件，将其导入Discovery软件，获得其3D结构。运用Discovery软件移除靶蛋白中的配体和非蛋白分子(如水分子)并进行加氢处理，然后在软件中对化合物和靶点蛋白进行分子对接，并进行可视化处理。Download the SDF format files of compounds 1-4 and positive control fluconazole, ketoconazole, and itraconazole structures from the Pub Chem database (https://pubchem.ncbi.nlm.nih.gov/), import them into the Discovery software, and obtain their 3D structures. Use the Discovery software to remove ligands and non-protein molecules (such as water molecules) in the target protein and perform hydrogenation treatment, then perform molecular docking and visualization of the compounds and target proteins in the software.

2.分子对接结果及分析2. Molecular docking results and analysis

Lib Dock Score指示了配体与靶点蛋白晶体的结合程度，Lib Dock Score越高，小分子与受体结合的活性越高。将化合物1-4分别与“1.1”中的蛋白质分别进行分子对接，并进行可视化处理，代表性结合图如图1至图4所示，分别展示了化合物4与1EQP、1CZ1、1AI9、4IIB进行分子对接的空间构型，以及化合物活性基团结合情况。Lib Dock Score见表9。Lib Dock Score indicates the degree of binding between the ligand and the target protein crystal. The higher the Lib Dock Score, the higher the activity of the small molecule binding to the receptor. Compounds 1-4 were molecularly docked with the proteins in "1.1" and visualized. Representative binding diagrams are shown in Figures 1 to 4, which respectively show the spatial configuration of compound 4 molecularly docked with 1EQP, 1CZ1, 1AI9, and 4IIB, as well as the binding of the active groups of the compounds. Lib Dock Score is shown in Table 9.

甾醇14-α脱甲基酶为多数唑类化合物潜在抗真菌靶点，但通过分子对接发现，与活性最优的酮康唑为对比，化合物1-4与甾醇14-α脱甲基酶(CYP51,PDB ID:5FSA和CYP51，PDB ID:5TZ1)的结合能力不强，而与葡聚糖酶(XOG1，PDB ID：1EQP、1CZ1和XOG1，PDB ID:1CZ1)、二氢叶酸还原酶(DFR1，PDB ID：1AI9)以及β-葡萄糖苷酶1(BGL1，PDB ID：4IIB)显示较高的结合能力。Sterol 14-α demethylase is a potential antifungal target for most azole compounds, but molecular docking revealed that, compared with the most active ketoconazole, compounds 1-4 had weak binding ability to sterol 14-α demethylase (CYP51, PDB ID: 5FSA and CYP51, PDB ID: 5TZ1), but showed higher binding ability to glucanase (XOG1, PDB ID: 1EQP, 1CZ1 and XOG1, PDB ID: 1CZ1), dihydrofolate reductase (DFR1, PDB ID: 1AI9) and β-glucosidase 1 (BGL1, PDB ID: 4IIB).

表9各化合物与靶点蛋白的Lib Dock Score
Table 9 Lib Dock Score of each compound and target protein

分子对接结果表明，与传统唑类化合物的作用靶点不同，所合成的活性化合物并非作用 于甾醇14-α脱甲基酶，而可能通过多靶点完成抗真菌作用，其抗真菌作用可能与葡聚糖酶、二氢叶酸还原酶以及β-葡萄糖苷酶1高度相关。The molecular docking results show that the synthesized active compounds are different from the targets of traditional azole compounds. Its antifungal effect may be highly related to glucanase, dihydrofolate reductase and β-glucosidase 1.

实施例8Example 8

制剂例(喷雾剂)Preparation example (spray)

一种皮毛动物外用喷雾剂(10mg/mL)，由以下原料按重量百分比制备而成：活性化合物1％、乙醇40％、甘油5％、硫代硫酸钠0.2％、水溶性氮酮0.5％、山梨酸0.2％、PVA0.5％、PVP 2％、1％吐温80。其中，活性化合物可以采用上述实施例制备的化合物1-20中的任意一种。A fur animal topical spray (10 mg/mL) is prepared from the following raw materials by weight percentage: 1% active compound, 40% ethanol, 5% glycerol, 0.2% sodium thiosulfate, 0.5% water-soluble azone, 0.2% sorbic acid, 0.5% PVA, 2% PVP, and 1% Tween 80. The active compound can be any one of the compounds 1-20 prepared in the above embodiments.

制备方法如下：按照上述原料配方比例取各种原料成分，将酒精置于烧杯中，加入甘油、硫代硫酸钠、水溶性氮酮、山梨酸，搅拌10min，混合均匀。然后缓慢加入PVA(聚乙烯醇，可乐丽PVA-217，下同)、PVP(聚乙烯吡咯烷酮，k30，下同)，充分搅拌，可适当加热以促进溶解，待PVA和PVP充分溶解后，加入吐温80，继续搅拌20min。冷却至50℃以下，加入活性化合物，搅拌至溶解充分，过滤，装瓶，制得皮毛动物外用喷雾剂。The preparation method is as follows: take various raw material ingredients according to the above raw material formula ratio, put alcohol in a beaker, add glycerol, sodium thiosulfate, water-soluble azone, sorbic acid, stir for 10 minutes, and mix evenly. Then slowly add PVA (polyvinyl alcohol, Kuraray PVA-217, the same below) and PVP (polyvinyl pyrrolidone, k30, the same below), stir well, and heat appropriately to promote dissolution. After PVA and PVP are fully dissolved, add Tween 80 and continue stirring for 20 minutes. Cool to below 50°C, add active compounds, stir until fully dissolved, filter, and bottle to obtain a fur animal external spray.

实施例9Example 9

制剂例(喷雾剂)Preparation example (spray)

一种皮毛动物外用喷雾剂(10mg/mL)，由以下原料按重量百分比制备而成：活性化合物1％、DMSO5％、10％吐温80、20％PEG400、乙醇20％、甘油5％、硫代硫酸钠0.2％、水溶性氮酮0.5％、山梨酸0.2％、PVA0.5％、PVP 2％。其中，活性化合物可以采用上述实施例制备的化合物1-20中的任意一种。A fur animal topical spray (10 mg/mL) is prepared from the following raw materials by weight percentage: 1% active compound, 5% DMSO, 10% Tween 80, 20% PEG400, 20% ethanol, 5% glycerol, 0.2% sodium thiosulfate, 0.5% water-soluble azone, 0.2% sorbic acid, 0.5% PVA, and 2% PVP. The active compound can be any one of the compounds 1-20 prepared in the above embodiments.

按照上述配方比例准备好原料，然后按照以下制备方法制成喷雾剂。Prepare the raw materials according to the above formula ratio, and then make the spray according to the following preparation method.

①称1g活性化合物，用5mL DMSO(二甲基亚砜)溶解，加入10mL吐温80，20mL PEG400(聚乙二醇，分子量380～420)，5mL甘油，20mL乙醇混合均匀。① Weigh 1g of active compound, dissolve it with 5mL DMSO (dimethyl sulfoxide), add 10mL Tween 80, 20mL PEG400 (polyethylene glycol, molecular weight 380-420), 5mL glycerol, and 20mL ethanol and mix well.

②向40mL水中加入0.2g硫代硫酸钠，0.2g山梨酸，0.5mL水溶性氮酮溶解后，一点点的加入2g PVP，0.5g PVA加热至溶解，冷却至室温。② Add 0.2g sodium thiosulfate, 0.2g sorbic acid, and 0.5mL water-soluble azone to 40mL water and dissolve them. Then add 2g PVP and 0.5g PVA little by little, heat until dissolved, and cool to room temperature.

③将②中溶解缓慢加入①中，逐渐搅拌至溶解，装瓶，制得皮毛动物外用喷雾剂。 ③ Slowly add the dissolved substance in ② into ①, gradually stir until dissolved, bottle, and obtain an external spray for fur animals.

Claims (10)

一种唑类化合物，其特征在于，具有式I所示分子结构的化合物：
An azole compound, characterized in that it has a molecular structure shown in Formula I:
其中，R1至少一个，R1各自独立的选自氢、C1-C4烷基、环己基、卤取代C1-C3烷基、C2-C4烯基、卤取代C2-C4烯基、硝基、氨基、卤素、甲氨基、羟基、羧基、羟基取代C1-C3烷基、苯基、甲基苯基、苯甲基、C1-C3烷氧基、C2-C4烯基氧基、磺酸基、硝基苯基、甲酰基、乙酰基、苯甲酰基、苄氧羰基、苄氧基、苯氧基、甲酰氧基、乙酰氧基、苯甲酰氧基、巯基、甲基磺酰基、磺酸基、C1-C3烷基氨基、双C1-C3烷基取代氨基或叔丁氧羰基；wherein, there is at least one R1 , and each R1 is independently selected from hydrogen, C1 - C4 alkyl, cyclohexyl, halogen-substituted C1- C3 alkyl, C2 - C4 alkenyl, halogen-substituted C2 - C4 alkenyl, nitro, amino, halogen, methylamino, hydroxyl, carboxyl, hydroxyl-substituted C1 - C3 alkyl, phenyl, methylphenyl, benzyl, C1 - C3 alkoxy, C2 - C4 alkenyloxy, sulfonic acid, nitrophenyl, formyl, acetyl, benzoyl, benzyloxycarbonyl, benzyloxy, phenoxy, formyloxy, acetoxy, benzoyloxy, mercapto, methylsulfonyl, sulfonic acid, C1 - C3 alkylamino, di- C1 - C3 alkyl-substituted amino or tert-butyloxycarbonyl; a表示R1的个数，a＝0～4；b表示R2的个数，b＝0～4；a represents the number of R1 , a=0-4; b represents the number of R2 , b=0-4; R2至少一个，R2各自独立的选自C1-C3烷基、卤素、氨基、甲氨基、二甲氨基、羟基、羧基、硝基、氰基或卤代烷基；At least one R 2 , each R 2 is independently selected from C 1 -C 3 alkyl, halogen, amino, methylamino, dimethylamino, hydroxyl, carboxyl, nitro, cyano or haloalkyl; X1为-(CH2)n-或-(CH2)n-O-或-O-(CH2)n-，n＝0～4； X1 is -( CH2 )n- or -( CH2 )nO- or -O-( CH2 )n-, n = 0 to 4; X2为-(CH2)m-或-C(O)-CH(CH3)-(CH2)m-，m＝0～4；X 2 is -(CH 2 )m- or -C(O)-CH(CH 3 )-(CH 2 )m-, m=0～4; Y为C或N。Y is C or N. 根据权利要求1所述一种唑类化合物，其特征在于，所述R1各自独立的选自：氢、甲基、乙基、正丙基、异丙基、异丁基、烯丙基、丙炔基、乙烯基、仲丁基、环己基、苯甲基、丙烯基、叔丁基、异丙烯基、乙炔基、苯基、对甲苯基、间甲苯基、邻甲苯基、邻硝基苯基、甲酰基、乙酰基、苯甲酰基、羧基、苄氧羰基、氨基、甲氨基、乙基氨基、丙基氨基、二甲氨基、二乙氨基、硝基、羟基、甲氧基、乙氧基、苄氧基、苯氧基、甲酰氧基、氟基、巯基、氯基、溴基、碘基、叔丁氧羰基或氟甲基。The azole compound according to claim 1, characterized in that the R 1 is independently selected from the group consisting of hydrogen, methyl, ethyl, n-propyl, isopropyl, isobutyl, allyl, propynyl, vinyl, sec-butyl, cyclohexyl, benzyl, propenyl, tert-butyl, isopropenyl, ethynyl, phenyl, p-tolyl, m-tolyl, o-tolyl, o-nitrophenyl, formyl, acetyl, benzoyl, carboxyl, benzyloxycarbonyl, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, nitro, hydroxyl, methoxy, ethoxy, benzyloxy, phenoxy, formyloxy, fluoro, thiol, chloro, bromo, iodo, tert-butoxycarbonyl or fluoromethyl. 根据权利要求1所述一种唑类化合物，其特征在于，所述R2各自独立的选自：C1-C3烷基、卤素、氨基、氟基、氯基、溴基、碘基、羟基、全氟C1-C3烷基；全氟C1-C3烷基是指全氟甲基、全氟乙基、全氟正丙基或全氟异丙基。An azole compound according to claim 1, characterized in that the R2 is independently selected from: C1 - C3 alkyl, halogen, amino, fluoro, chloro, bromo, iodo, hydroxyl, perfluoro C1 - C3 alkyl; perfluoro C1 - C3 alkyl refers to perfluoromethyl, perfluoroethyl, perfluoro-n-propyl or perfluoro-isopropyl. 根据权利要求1所述一种唑类化合物，其特征在于，a表示R1的个数，a＝0～3。The azole compound according to claim 1, characterized in that a represents the number of R1 , and a=0-3. 根据权利要求1所述一种唑类化合物，其特征在于，b表示R2的个数，b＝0～3。The azole compound according to claim 1, characterized in that b represents the number of R2 , and b=0-3. 根据权利要求1所述一种唑类化合物，其特征在于，n＝0～3。 The azole compound according to claim 1, characterized in that n=0-3. 根据权利要求1所述一种唑类化合物，其特征在于，m＝0～3。The azole compound according to claim 1, characterized in that m=0-3. 根据权利要求1所述一种唑类化合物，其特征在于，所述唑类化合物是具有式II或式III所示分子结构的化合物：
An azole compound according to claim 1, characterized in that the azole compound is a compound having a molecular structure shown in Formula II or Formula III:
其中，n＝0-3；Wherein, n = 0-3; Y为C或N；Y is C or N; R1至少一个，R1各自独立的选自氢、C1-C4烷基、环己基、卤取代C1-C3烷基、C2-C4烯基、卤取代C2-C4烯基、硝基、氰基、氨基、卤素、甲氨基、羟基、羧基、羟基取代C1-C3烷基、苯基、甲基苯基、苯甲基、C1-C3烷氧基、C2-C4烯基氧基、磺酸基、硝基苯基、甲酰基、乙酰基、苯甲酰基、苄氧羰基、苄氧基、苯氧基、甲酰氧基、乙酰氧基、苯甲酰氧基、巯基、甲基磺酰基、磺酸基、C1-C3烷基氨基、双C1-C3烷基取代氨基或叔丁氧羰基； R1 is at least one, and R1 is independently selected from hydrogen, C1 - C4 alkyl, cyclohexyl, halogen-substituted C1 - C3 alkyl, C2 - C4 alkenyl, halogen-substituted C2 - C4 alkenyl, nitro, cyano, amino, halogen, methylamino, hydroxy, carboxyl, hydroxy-substituted C1 - C3 alkyl, phenyl, methylphenyl, benzyl, C1 - C3 alkoxy, C2 - C4 alkenyloxy, sulfonic acid, nitrophenyl, formyl, acetyl, benzoyl, benzyloxycarbonyl, benzyloxy, phenoxy, formyloxy, acetoxy, benzoyloxy, mercapto, methylsulfonyl, sulfonic acid, C1 - C3 alkylamino, di- C1 - C3 alkyl-substituted amino or tert-butyloxycarbonyl; a表示R1的个数，a＝0～4；b表示R2的个数，b＝0～4；a represents the number of R1 , a=0-4; b represents the number of R2 , b=0-4; R2至少一个，R2各自独立的选自C1-C3烷基、卤素、氨基、甲氨基、二甲氨基、羟基、羧基、硝基、氰基或卤代烷基。There is at least one R 2 , and each R 2 is independently selected from C 1 -C 3 alkyl, halogen, amino, methylamino, dimethylamino, hydroxyl, carboxyl, nitro, cyano or halogenated alkyl. 根据权利要求1所述一种唑类化合物，其特征在于，所述唑类化合物是以下化合物中的一种：

The azole compound according to claim 1, characterized in that the azole compound is one of the following compounds:

权利要求1-9任意一项所述唑类化合物在制备药物中的应用。 Use of the azole compound according to any one of claims 1 to 9 in the preparation of medicines.
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