Compounds modified by bensulfuron methyl and application of compounds in preparation of antifungal drugs

Abstract

The invention relates to a bensulfuron methyl modified compound and application of the bensulfuron methyl modified compound in preparation of drugs for resisting infection caused by fungi, in particular to application of the bensulfuron methyl modified compound in preparation of drugs for resisting infection caused by candida auricula. The inhibition constant Ki value of the compound disclosed by the invention on candida auricula AHAS can reach 0.33 mu M.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and relates to a class of compounds modified by benzylsulfuron-methyl and uses thereof in preparing drugs for resisting infections caused by fungi, and specifically relates to a class of compounds modified by benzylsulfuron-methyl and uses thereof in preparing drugs for resisting infections caused by Candida auris. Technical Background

[0002] According to the Global Action on Fungal Infections (https: / / gaffi.org / why / fungal-disease-frequency / ), more than 300 million people suffer from serious fungal infections, and the health of 25 million patients is at great risk. The types of antifungal drugs used to treat such diseases are very limited. The families of antifungal drugs that can be used clinically to treat such diseases 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 first officially reported in the external auditory canal of a hospitalized patient in Japan (Satoh, K., et al., Microbiol. Immunol., 2009, 53(1), 41-44), which was resistant to fluconazole. This deadly fungus has now been found in more than 45 countries on six continents and is known as a super fungus.

[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 2011, Wang Jianguo et al. (CN10248862A) found that commercial sulfonylurea herbicides such as chlorimuron-methyl, ethoxysulfuron-methyl and benzylsulfuron-methyl had strong in vitro inhibition of Candida albicans AHAS and showed strong inhibitory effects on Candida albicans, and patented the biological activities of these compounds. In 2020, Professor Luke Guddat's team at the University of Queensland, Australia, discovered that benzylsulfuron-methyl showed strong inhibition of Candida auris AHAS, and showed the strongest inhibitory effect on Candida auris strains in terms of 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. It has great prospects. With the rational modification of benzylsulfuron-methyl, it is possible to discover new sulfonylurea compounds with higher antifungal activity.

[0004] Summary of the invention

[0005] The object of the present invention is to provide a class of compounds modified from bensulfuron-methyl and their use in preparing drugs for treating infections caused by fungi, especially their use in preparing drugs for treating infections caused by Candida auris. The class of compounds modified from bensulfuron-methyl of the present invention is

[0006] and its medicinal salts.

[0007] A new type of sulfonylurea compound of the present invention is obtained by the following reaction formula:

[0008] (1) For compounds 1 to 12

[0009]

[0010] (2) For compounds 13 to 24

[0011] Among them, the synthetic route of intermediate D is

[0012]

[0013] The present invention also provides a drug for treating fungal infections in humans or animals, especially Candida auris, Candida albicans, Cryptococcus neoformans, Saccharomyces cerevisiae, Candida parapsilosis, and Candida glabrata. An effective amount of a benzene alkoxy-substituted sulfonylurea compound is applied to the fungus or its environment to kill the fungus or control fungal growth. The drug may contain the above-mentioned benzene alkoxy-substituted sulfonylurea compound and one or more pharmaceutically acceptable carriers. The carriers include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, and synergists in the pharmaceutical field. The drug can be used in the form of injections, tablets, pills, capsules, suspensions, or emulsions. The administration route can be oral, transdermal, intravenous, or intramuscular for treating fungal infections in humans or animals. Brief Description of the Drawings

[0014] Figure 1 It is the inhibition curve of compound 8 against Candida auris AHAS. Detailed Embodiments

[0015] The substantial features of the present invention can be embodied in the following examples, but these examples are only for illustration and do not limit the present invention.

[0016] Example 1. Synthesis of Intermediate D

[0017] The synthesis of intermediate M was completed according to the method of the reference (Sun, X.W., et al., Chem. Biol. Drug Des. 2024, 103(1): e14364).

[0018] Intermediate M (975 mg, 2.80 mmol) was dissolved in 30 mL of ethyl acetate, and the solution was cooled in an ice bath. tert-Butylamine (409 mg, 5.60 mmol) was slowly added to the solution. The resulting white suspension was stirred at 0 °C for 1 hour and continued to be stirred at room temperature for 2 hours. The mixture was washed with 30 mL of 5% hydrochloric acid. The organic layer was dried over anhydrous magnesium sulfate and evaporated under reduced pressure to obtain 560 mg of compound N as a pale yellow oil, with a yield of 52%. 1H NMR (400 MHz, CDCl 3 )δ8.09-8.02(m,1H,ArH),7.62-7.40(m,3H,ArH),4.91(s,2H,CH 2 SO 2 ), 4.78(s, 2H, OCH 2 CO), 1.53(s, 9H, OC(CH 3 ) 3 ), 1.21(s, 9H, NHC(CH 3 ) 3 ).

[0019] Intermediate N (188 mg, 0.49 mmol) was added to 1 mL of trifluoroacetic acid. The mixture was stirred at room temperature for 2 hours and then evaporated to dryness. Water (30 mL) was added to the residue and the mixture was extracted with ethyl acetate (2×30 mL). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated on a rotary evaporator. The residue was purified by using a chromatograph using an ethyl acetate / n-hexane mixture (1:1, v / v) as an eluent to give 57 mg of a yellow solid D with a yield of 43%. 1 H NMR (400 MHz, DMSO-d 6 )δ13.01 (s, 1H, COOH), 7.85 (dd, J=7.8, 1.5Hz, 1H, ArH), 7.56 (td, J=7.5, 1.5Hz, 1H, ArH), 7.45 (ddd, J=9.0, 7.1, 1.5Hz, 2H, ArH), 6.85 (s, 2H, OCH 2 CO), 4.87 (d, J=1.8Hz, 2H, CH 2 SO 2 ).

[0020] Example 2. Synthesis of Compound 8

[0021] Intermediate A (Cas number 690638-32-5) was purchased from Arctom Product List, and the synthesis of intermediate B was completed according to the method in the reference (Sun, XW, et al., Chem. Biol. Drug Des. 2024, 103(1): e14364).

[0022] To a stirred solution of intermediate B (100 mg, 0.26 mmol) in 20 mL of acetonitrile was added 2-bromoacetamide (53 mg, 0.39 mmol) and DBU (79 mg, 0.52 mmol). The reaction mixture was heated at 50°C for 1 hour, then concentrated on a rotary evaporator, acidified with 30 mL of 5% hydrochloric acid, and extracted with ethyl acetate. The combined organic layers were dried over anhydrous magnesium sulfate.

[0023] The solvent was removed on a rotary evaporator and the residue was purified twice by preparative TLC on silica gel using ethyl acetate:dichloromethane=1:1 as a developing solvent to obtain 50 mg of a white solid, which was Compound 8.

[0024] Example 3. Synthesis of Compound 20

[0025] Add 109.2 mg of intermediate D (0.4 mmol) to a 25 mL round-bottom flask containing 5 mL of acetonitrile, add 121 mg of phenyl (4, 6-dimethoxypyrimidin 2-yl) carbamate (0.44 mmol) after dissolving, stir evenly and drop 65 μL of DBU (0.44 mmol), stir overnight at room temperature. After sufficient time, pour the reaction solution into a conical flask, add 10 mL of water, adjust the pH value to about 2 with 5% HCl, and precipitate begins to precipitate. Let stand until the precipitation is complete. Filter, wash with water, and dry to obtain a white solid powder, which is the target compound.

[0026] Properties, yields, melting points, 1 H NMR, 13 C NMR and HRMS data are shown in Table 1.

[0027] Table 1. Physicochemical characterization and spectral data of compounds 1 to 24

[0028]

[0029]

[0030]

[0031]

[0032]

[0033] Example 4. Determination of the inhibitory activity of compounds against AHAS of Bacillus auris

[0034] 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).

[0035] 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).

[0036] Inhibition constant K i Calculated by the following formula

[0037]

[0038] 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.

[0039] The inhibitory activities of the tested compounds against C. auris AHAS are shown in Table 1.

[0040] Table 2. Inhibition rate of compound 1-compound 24 on AHAS of C. auris at a concentration of 50 μM (%)

[0041] Compound No. Inhibition rate Compound No. Inhibition rate Compound No. Inhibition rate Compound 1 0 Compound 10 29 Compound 19 12 Compound 2 0 Compound 11 32 Compound 20 61 Compound 3 0 Compound 12 94 Compound 21 0 Compound 4 0 Compound 13 0 Compound 22 24 Compound 5 0 Compound 14 0 Compound 23 0 Compound 6 0 Compound 15 0 Compound 24 64 Compound 7 0 Compound 16 0 Bensulfuron-methyl 91 Compound 8 97 Compound 17 0 Compound 9 0 Compound 18 0

[0042] It can be seen that the compounds of the present invention have good inhibition on AHAS of Candida auris, especially compound 8 and compound 12, whose inhibition rate on AHAS of Candida auris is higher than that of the control drug benzylsulfuron-methyl.

[0043] Table 3. Inhibition constants of some compounds against AHAS of Escherichia coli

[0044] Compound No. <![CDATA[K i (μMM)]]> Compound 8 0.33±0.11 Compound 12 4.38±0.61 Bensulfuron-methyl 0.61±0.09

[0045] It can be seen that the inhibition constants K of compounds 8 and 12 of the present invention for Candida auris AHAS are i All of them are relatively strong, especially compound 8, whose inhibition constant K i It is nearly twice as high as the control drug benzylsulfuron-methyl.

[0046] According to the research results of Sun XW et al. (Sun, XW, et al, Chem. Biol. Drug Des. 2024, 103, e14364.), for the analogs of benzsulfuron-methyl, their inhibitory intensity against Candida auris AHAS and their anti-Auris strain effect in cell models are highly consistent. Therefore, it can be reasonably inferred that under the same conditions, the inhibitory effect of compound 8 on Candida auris in the cell model should be stronger than the inhibitory effect of benzsulfuron-methyl on Candida auris.

Claims

1. A compound modified from benzylsulfuron-methyl, characterized in that The compound modified by benzylsulfuron-methyl is Its medical salt.

2. Use of the compound modified from bensulfuron-methyl as claimed in claim 1 in the preparation of antifungal drugs.

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

4. The use according to claim 2, characterized in that The antifungal drug contains the above-mentioned compound modified from bensulfuron-methyl 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.

5. The use according to claim 2, characterized in that The antifungal drug can be used in the form of injection, tablet, pill, capsule, suspension or emulsion.

6. The use according to claim 2, characterized in that The antifungal drug can be administered orally, percutaneously, intravenously or intramuscularly.

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