阅读说明：本技术 3-苯基吲哚衍生物制备抗真菌药物的应用 (Application of 3-phenylindole derivative in preparation of antifungal drugs ) 是由 倪广惠 赵胤 王睿睿 殷珊 孙爱梅 吴艳丹 张祎 刘瑞娜 于 2021-08-13 设计创作，主要内容包括：本发明公开了3-苯基吲哚衍生物制备抗真菌药物的应用。通过实验证明,3-苯基吲哚衍生物单独使用和联合抗真菌药物具有显著抑制耐药白色念珠菌的作用。本发明提供的应用具有很好的药用价值与应用前景。(The invention discloses application of 3-phenylindole derivatives in preparation of antifungal drugs. Experiments prove that the 3-phenylindole derivative has the function of obviously inhibiting drug-resistant candida albicans when being used alone or combined with antifungal drugs. The application provided by the invention has good medicinal value and application prospect.)

1.3-phenyl indole derivatives for preparing antifungal drugs, which is characterized in that the structural formula of the 3-phenyl indole derivatives is shown in the specification(ii) a Wherein R = H, OCH₃。

2. Use according to claim 1, characterized in that the fungus is Candida albicans, Marneffei staphylium, Cryptococcus and Malassezia furfur.

3. The antifungal pharmaceutical composition of claim 1 wherein the fungus is a Candida albicans resistant strain, a Marneffei staphylium resistant strain, a Cryptococcus resistant strain, a Malassezia furfur resistant strain.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of a 3-phenylindole derivative in preparation of an antifungal medicine.

Background

Invasive mycosis refers to systemic infectious diseases caused by invasion of tissues, organs and blood by various pathogenic candida, and the most common pathogenic bacteria include candida albicans, candida tropicalis, candida glabrata and candida kininogenes. Most life-threatening fungal infections occur in immunocompromised trauma, high immunosuppressive usage, neutropenia, medical device implantation and organ transplant patients with a mortality rate of about 40%. With the intervention treatment of antibacterial drugs such as fluconazole, the clinical isolate produces drug resistance genes and forms cross resistance with other azole drugs. Making the treatment of invasive mycoses problematic. In the screening of medicines, the inventor unexpectedly finds that the 3-phenylindole derivative has a good synergistic antibacterial effect on drug-resistant candida albicans, which is the first discovery.

Disclosure of Invention

The invention aims to provide application of 3-phenylindole derivatives in preparing antifungal medicines, wherein the structural formula of the 3-phenylindole derivatives is shown in the specification(ii) a Wherein R = H, OCH₃。

The fungi are Candida albicans, Marneffei basket fungus, Cryptococcus and Malassezia furfur.

The fungi are Candida albicans resistant strain, Marneffei basket fungus resistant strain, Cryptococcus resistant strain, and Malassezia furfur resistant strain.

The invention has the advantages that:

the invention has the advantages and positive effects that sufficient chemical and pharmacological basis is provided for drug candidates for treating drug-resistant deep fungal infections.

Detailed Description

The present invention is further illustrated but not limited in any way by the following examples, and any modifications made thereto are intended to fall within the scope of the present invention.

The application of 3-phenylindole derivatives in preparing antifungal medicines, wherein the structural formula of the 3-phenylindole derivatives is shown in the specification(ii) a Wherein R = H, OCH₃。

The fungi are Candida albicans, Marneffei basket fungus, Cryptococcus and Malassezia furfur.

The fungi are Candida albicans resistant strain, Marneffei basket fungus resistant strain, Cryptococcus resistant strain, and Malassezia furfur resistant strain.

Example 1

Indole derivatives antifungal activity test:

materials and methods

Medicine and sample

The preparation method of the indole derivative comprises the following steps:

synthesis of 3-phenyl-5-methoxyindole

In a dry round bottom flask was added 5-methoxyindole (3.0 mmol, 442.0 mg) and potassium tert-butoxide (6.0 mmol, 694.0 mg), the solid mixture was vacuum/nitrogen purged three times, then 6.5 mL of degassed dimethyl sulfoxide was injected (freeze-pump-thaw), the solid was dissolved after 5 min of stirring, and iodobenzene (1.5 mmol, 171 uL) was added dropwise, stirring at 80 ℃ under nitrogen. After reacting for 40 h, adding 6 mL of deionized water for quenching, extracting with 60 mL of ethyl acetate, then washing with half-saturated sodium chloride solution and saturated sodium chloride solution for 3 times in sequence, and drying the organic layer with anhydrous sodium sulfate. Filtration was carried out, the solvent was evaporated under reduced pressure from the filtrate, and the residue was purified by silica gel column chromatography to give 162.2 mg (yield: 24%) of 3-phenyl-5-methoxyindole.

Synthesis of 3-phenylindoles

Indole (1.0 mmol, 120.0 mg) and potassium tert-butoxide (2.0 mmol, 231.4 mg) were added to a dry round bottom flask, the solid mixture was vacuum/nitrogen purged three times, then 1.5 mL degassed dimethyl sulfoxide was injected (freeze-pump-thaw), the solid was dissolved after 5 min of stirring, and iodobenzene (0.5 mmol, 51 uL) was added dropwise with stirring at 80 ℃ under a nitrogen atmosphere. After 42 h of reaction, the reaction was complete as checked by TLC. Quenched by the addition of 2 mL of deionized water, extracted with 20 mL of ethyl acetate, then washed 3 times with half-saturated sodium chloride solution, and the organic layer dried over anhydrous sodium sulfate. Filtration was carried out, the solvent was distilled off from the filtrate under reduced pressure, and the residue was purified by silica gel column chromatography to give 47 mg of 3-phenylindole (yield 24.3%).

The sample 3-phenylindole derivative was dissolved at 100mg/ml and placed in a refrigerator at 4 ℃. Grinding fluconazole, dissolving with DMSO, ultrasonic treating for 10min, centrifuging to obtain supernatant, storing at 50mg/ml, and placing in refrigerator at 4 deg.C.

II, reagent and solution

(1) Reagent

Fluconazole, DMSO.

(2) Culture medium

Sabouraud broth.

Third, Experimental methods

Taking a 96-well culture plate, diluting the positive drug fluconazole into an initial concentration of 200ug/ml, directly using a storage solution for a sample, directly adding 200ug/ml fluconazole into the storage solution for the sample during drug combination, then diluting by 5 times, carrying out 4 concentration gradients, and repeating 3 wells for each concentration gradient. Adding fungus suspension into each well, wherein the concentration of Candida albicans standard strain is 1 × 10⁵CFU/mL, concentration of Candida albicans resistant strain 1 × 10⁵ CFU/mL, cultured at 37 ℃ for 24 h. The OD value at 630nm was measured by a microplate reader. The experiment was also set with a medium blank control, a bacteria solution control and a fluconazole positive drug control.

Fourth, calculation formula

Fungal activity inhibition (%) = (1-sample OD value/Experimental control hole OD value) × 100%

Combination Index (FICI): FICI = MICA/A + MICB/B (where A and B are MIC values for single use of the two drugs, respectively, and MICA and MICB are MIC values for combination of the two drugs, respectively.) when the MIC values are higher than the detection upper limit, a value twice the upper limit concentration is used to calculate the FICI. the MIC50 value in this experiment is calculated as the percentage inhibition of fluconazole at 50% minimal fungal growth. when FICI is less than or equal to 0.5, the two drugs act in a synergistic manner, when FICI is greater than 0.5 and less than or equal to 4, the two drugs act in an unrelated manner, and when FICI is greater than 4, the two drugs act in an antagonistic manner.

The results of the experiments are shown in the following table.

The experimental result shows that the indole derivative has antibacterial effect on drug-resistant bacteria; the compound has the function of inhibiting candida albicans drug-resistant bacteria when being used together with the antifungal drug fluconazole, and has strong synergistic effect.