Application of benzene ring alkoxy substituted sulfonylurea compound in preparation of antifungal drugs

Abstract

The invention relates to an application of a benzene ring alkoxy substituted sulfonylurea compound in preparation of a medicine for resisting infection caused by fungi, in particular to an application in preparation of a medicine for resisting infection caused by candida auricula. The inhibition constant Ki value of the compound to the candida auricula AHAS can reach 110 nM, and the minimum half inhibition concentration of the compound to the candida auricula can reach 1.2 mu M.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and relates to the use of a class of phenylcycloalkoxy substituted sulfonylurea compounds in preparing drugs for preventing fungal infections, and specifically relates to the use of a class of phenylcycloalkoxy substituted sulfonylurea compounds in preparing drugs for preventing infections caused by Candida auris. Technical Background

[0002] Infections caused by various invasive fungi have a serious impact on human health, such as tinea pedis, gynecological urinary tract and vaginitis, AIDS caused by surgical wound infections, etc. The types of clinical drugs used to treat fungal infections are very limited, including azoles, echinocandins, polyenes, allylamines and pyrimidine analogs, among which amphotericin B (AMB) and fluconazole (FCZ) are the most widely used drugs. Due to the overuse of these antifungal drugs, drug resistance has become a serious problem. In 2009, Candida auris was officially reported for the first time in the external auditory canal of a hospitalized patient in Japan (Satoh, K., et al., Microbiol. Immunol., 2009, 53 (1), 41-44). It showed resistance to antifungal drugs such as fluconazole and was called super fungus. This deadly fungus has been found in more than 45 countries on six continents.

[0003] Acetolactate synthase (acetohydroxyacid synthase, AHAS, EC 2.2.1.6) is the first key enzyme that catalyzes the biosynthesis of three branched-chain amino acids such as valine. This life process does not exist in mammals, so inhibitor drugs targeting AHAS are biosafe for humans (Duggleby, RG, et al., J.Biochem.Mol.Biol.2000, 33, 1-36). In 2020, Professor Luke Guddat's team at the University of Queensland, Australia, found that benzylsulfuron-methyl showed strong inhibition of Candida auris AHAS, and showed the strongest inhibitory effect on Candida auris strains in cell activity (Agnew-Francis, KA, et al., ACS Infect.Dis.2020, 6, 2901-2912). The above progress shows that targeting AHAS to design antifungal drugs is a relatively novel idea, especially for infections caused by super fungi such as Candida auris, and has great prospects.

[0004] Wang Jianguo et al. recently designed and synthesized a series of phenylcycloalkoxy-substituted sulfonylurea compounds, which showed extremely high control effects on certain difficult-to-control grass weeds in farmland (CN110642791 B and CN117903066 A).

[0005] Summary of the invention

[0006] The object of the present invention is to provide a type of benzene ring alkoxy substituted sulfonylurea compound for use in preparing a drug for antifungal infection, especially for preparing a drug for infection caused by Candida auris. The type of benzene ring alkoxy substituted sulfonylurea compound has a strong inhibitory effect on Candida auris AHAS and has a strong inhibitory effect on Candida auris in a cell model.

[0007] The present invention provides a class of benzene ring alkoxy substituted sulfonylurea compounds:

[0008] and its medicinal salts.

[0009] A new type of sulfonylurea compound of the present invention is obtained by the following reaction formula:

[0010]

[0011] The specific process and details have been introduced in detail in patents CN110642791B and CN117903066A.

[0012] The present invention also provides a drug for treating fungal infections in humans or animals, in particular Candida auris, Candida albicans, Cryptococcus neoformans, Saccharomyces cerevisiae, Candida parapsilosis and Candida glabrata, by applying an effective amount of a phenylcycloalkoxy substituted sulfonylurea compound to the fungus or its environment to kill the fungus or control the growth of the fungus. The drug may contain the above-mentioned phenylcycloalkoxy substituted sulfonylurea compound and one or more pharmaceutically acceptable carriers. The carrier includes conventional diluents, excipients, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and synergists in the pharmaceutical field. The drug can be prepared into an injection, tablet, pill, capsule, suspension or emulsion for use. Its administration route can be oral, percutaneous, intravenous or intramuscular injection, and is used to treat fungal infections in humans or animals. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 is the inhibition curve of compound 2022-LS5 on Candida auris AHAS, Figure 2This is the inhibition curve of compound 2022-LS5 against C. auris strain CBS10913 (the control agents are fluconazole and benzylsulfuron-methyl). DETAILED DESCRIPTION

[0014] The essential features of the present invention can be reflected in the following embodiments, but these embodiments are only for illustration rather than limitation of the present invention.

[0015] Example 1. Preparation of Compound 2022-LA5

[0016] The preparation method was prepared according to the method of patent CN117903066A. 105 mg (0.44 mmol) of 2-(2,2-difluoroethoxy)benzenesulfonamide was dissolved in 5 mL of acetonitrile, and 103 mg (0.4 mmol) of (4-methoxy-6-methylpyrimidin-2-yl)phenylcarbamate and 65 μL (0.44 mmol) of DBU (1,8-diazabicyclo[5.4.0]undec-7-ene) were added, and stirred at room temperature overnight. Then 10 mL of water was added to the reaction solution, and the pH was adjusted to 2 with 5% hydrochloric acid. A white solid was precipitated, and 96.5 mg of the product 2022-LA5 was obtained after filtration and drying, with a yield of 46%.

[0017] Likewise, compounds CMO, FMO and 2022-LS5 can be synthesized.

[0018] Example 2. Determination of the inhibitory activity of compounds against AHAS of Bacillus auris

[0019] The constructed plasmid of the catalytic subunit of Candida auris AHAS was transferred into Escherichia coli BL21, and its expression was induced by IPTG, and then the protein was purified by immobilized metal affinity chromatography IMAC. The purified protein was stored in a -80°C refrigerator to maintain its enzyme activity (Agnew-Francis, KA, et al., ACS Infect. Dis. 2020, 6, 2901-2912).

[0020] When the enzyme inhibition activity was measured, the solution contained 200mM phosphate buffer solution (pH 7.2), 100mM sodium pyruvate, 10mM magnesium chloride, 1mM ThDP, 10μM FAD and different concentrations of inhibitors. The enzyme activity was measured by continuous pyruvate consumption method, the reaction temperature was 30℃, and the OD was monitored in real time. 333 changes (Sun, XW, et al., Chem. Biol. Drug Des. 2024, 103(1): e14364).

[0021] Inhibition constant K i Calculated by the following formula

[0022]

[0023] Among them, V max represents the maximum catalytic reaction rate when the AHAS enzyme is not inhibited, [I] represents the concentration of the compound, and V represents the reaction rate.

[0024] The inhibitory activities of the tested compounds against C. auris AHAS are shown in Table 1.

[0025] Table 1. Inhibition constants of test compounds and control drugs against Candida auris AHAS

[0026] Compound No. <![CDATA[K of Candida auris AHAS i (nM)]]> CMO 420±70 FMO 210±30 2022-LA5 190±50 2022-LS5 110±30 Bensulfuron-methyl 613±88

[0027] As can be seen from Table 1, the inhibition constant K of the compounds of the present invention for Candida auris AHAS i Both were lower than that of the control drug benzylsulfuron-methyl, indicating that it had stronger activity in inhibiting the isolated AHAS enzyme.

[0028] Example 3. Inhibitory activity of compounds against Candida auris

[0029] The Candida auris used in the experiment came from the Westerdijk Fungal Biodiversity Institute in the Netherlands, with the code names CBS10913 and CBS12373. CBS10913 is a drug-sensitive strain isolated from a Japanese patient, and CBS12373 is a strain from South Korea that is resistant to azole antibiotics. In this experiment, the broth dilution method was used to determine the anti-candida auris activity of cell activity, and yeast nitrogen base (YNB) medium was selected for the study.

[0030] Inocula of CBS10913 and CBS12373 were prepared by resuspending colonies picked from 48-hour cultures grown on YPD agar in approximately 5.0 mL of sterile Milli-Q water. The cell concentration of the solution was measured using a hemacytometer and adjusted to (1.5-2.0) × 10 cells / mL with YNB broth. 6 CFU / mI and further diluted 1:1000 with YNB. ​​A 100 μL aliquot was then added to each well of the above 96-well plate (excluding inhibitor and medium-only controls) to a total volume of 200 μL per well. The plates were then incubated at 35°C for 48 hours and the OD at 48 hours was recorded. 525 .

[0031] MIC 50The MIC refers to the concentration of the test compound that inhibits the growth of C. auris by 50%, while the MIC refers to the minimum concentration of the test compound that completely inhibits the growth of C. auris. Cell-based inhibition data were obtained using a CLARIOstar multi-mode microplate reader (BMGlabtech).

[0032] Three parallel measurements were performed at each concentration.

[0033] The inhibitory activities of the tested compounds against Candida auris are shown in Table 2.

[0034] Table 2 Minimum inhibitory concentrations of test compounds and control drugs against two species of Candida auris

[0035]

[0036] It can be seen that the compounds of the present invention have a good inhibitory effect on Candida auris in the cell model, especially the compound 2022-LS5, which has a MIC of 0.040 for two strains of Candida auris. 50 The values ​​were lower than those of the control drugs fluconazole and benzylsulfuron-methyl.

Claims

1. Use of a class of phenylcycloalkoxy substituted sulfonylurea compounds in the preparation of antifungal drugs, characterized in that The benzene ring alkoxy substituted sulfonylurea compound is and its medicinal salts.

2. The use according to claim 1, characterized in that The fungi are Candida auris, Candida albicans, Cryptococcus neoformans, Saccharomyces cerevisiae, Candida parapsilosis and Candida glabrata.

3. The use according to claim 1, characterized in that The antifungal drug contains the above-mentioned benzene ring alkoxy substituted sulfonylurea compound and one or more pharmaceutically acceptable carriers, which include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and synergists in the pharmaceutical field.

4. The use according to claim 1, characterized in that The antifungal drug can be used in the form of injection, tablet, pill, capsule, suspension or emulsion.

5. The use according to claim 1, characterized in that The antifungal drug can be administered orally, percutaneously, intravenously or intramuscularly.

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