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Pharmaceutical composition capable of killing drug-resistant aspergillus fumigatus and method for killing drug-resistant aspergillus fumigatus

A technology of drug-resistant Aspergillus fumigatus and composition, applied in the field of microorganisms, can solve problems such as long

Inactive Publication Date: 2015-04-01
GUANGXI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

How to control the IPA caused by drug-resistant Aspergillus fumigatus has become a clinical problem. Developing new and effective drugs is a way to solve this problem. However, developing new and effective drugs is a long process, and it is difficult to deal with current drug resistance. severe situation
If a certain drug can cooperate with traditional antifungal drugs to make the latter play a stronger role in inhibiting Aspergillus fumigatus, it will also be of great help to alleviate the pressure of drug resistance, but no similar reports have been seen so far

Method used

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  • Pharmaceutical composition capable of killing drug-resistant aspergillus fumigatus and method for killing drug-resistant aspergillus fumigatus
  • Pharmaceutical composition capable of killing drug-resistant aspergillus fumigatus and method for killing drug-resistant aspergillus fumigatus
  • Pharmaceutical composition capable of killing drug-resistant aspergillus fumigatus and method for killing drug-resistant aspergillus fumigatus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Drug-resistant Aspergillus fumigatus pharmaceutical composition test

[0022] (1) Sources of strains Seven strains of Aspergillus fumigatus (numbered as AF 5, AF 14, AF 15, AF 17, AF 18, AF 26, AF 32; Among them, 5 strains of sputum samples, 1 strain of alveolar lavage fluid, and 1 strain of wound secretion) were identified as classic Aspergillus fumigatus by lactic acid phenol cotton blue staining morphology and 48°C temperature test. Sex tests confirmed that all strains were resistant to Amphotericin B (AMB). The quality control strain was Candida parapsilosis ATCC22019 specified by CLSI M38-A2 guidelines.

[0023] (2) Activation of strains and preparation of spore suspension Resuscitate 7 clinical strains of AMB drug-resistant Aspergillus fumigatus and inoculate them on a PDA slope, incubate at 35-37 °C for 3-5 days, and wash the surface of the slope with PBS solution containing 0.025% Tween 20 Collect the spores, resuspend the spores in RPMI...

Embodiment 2

[0031] Embodiment 2 pharmaceutical composition test result

[0032] (1) Quality control standard The MIC value of the quality control strain ATCC 22019 to AMB measured by each independent experiment is 1 μg / mL, which is within the reference range specified in the CLSI M38-A2 guideline, and the strains in the growth control wells grow well. Allergy test results are reliable.

[0033] (2) MIC of citral and AMB used alone or in combination According to the ECVs standard recommended by CLSI, the sensitivity of the tested Aspergillus fumigatus to AMB was judged, and the MIC of AMB was considered as sensitive and >1 μg / mL as resistant. When AMB was used alone, the MIC range of the tested strains was 2-4 μg / mL, that is to say, all the 7 strains of Aspergillus fumigatus tested in this experiment were resistant to AMB. The MIC value of citral alone is 256~512μg / mL. See Table 1 for details.

[0034] Table 1 MIC values ​​of citral and AMB alone or in combination on 7 strains of AMB-re...

Embodiment 3

[0040] Example 3 Cytotoxicity Experiment

[0041] Take 1-3ml of heparin anticoagulant blood from healthy adults, wash it with PBS buffer at pH=7.0 for 3 times, and centrifuge at 20-25°C and 2500rpm for 10min, remove the centrifuged supernatant, add PBS buffer, and prepare 10% erythrocyte-PBS suspension, and then dilute the suspension 1:10 with PBS, take 500 μL and place it in an EP tube, add 500 μL of citral or positive control amphotericin B serially diluted twice in PBS. Incubate the EP tube at 37°C for 1 h, centrifuge at 20-25°C and 2500 rpm for 10 min, transfer 150 μL of the supernatant to a 96-well plate, and measure the absorbance A at a wavelength of 540 nm with a microplate reader 540 . Red blood cells treated with 0.1% Triton X-100 in Triton X-100 wells were regarded as 100% complete hemolysis, and negative control wells were treated with red blood cells-PBS suspension containing 1% DMSO as 0% hemolysis. Hemolysis percentage (%) = (citral or amphotericin B hole A 5...

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Abstract

The invention discloses a pharmaceutical composition capable of killing drug-resistant aspergillus fumigatus and a method for killing drug-resistant aspergillus fumigatus, and belongs to the technical field of microorganism. The pharmaceutical composition is prepared from citral and an antifungal drug, wherein the antifungal drug comprises amphotericin B, voriconazole and itraconazole. The method for killing drug-resistant aspergillus fumigatus comprises the steps of preparing the composition, preparing a spore suspension, determining the drug MIC (Minimal Inhibitory Concentration) and carrying out a united drug sensitivity test. The pharmaceutical composition disclosed by the invention improves the antibacterial effect of the drug to the drug-resistant strain, reduces the toxic or side effects of the antifungal drug and is an ideal pharmaceutical composition for treating antifungal drug resistant aspergillus fumigatus infection.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a pharmaceutical composition for killing drug-resistant Aspergillus fumigatus and a method for killing drug-resistant Aspergillus fumigatus. Background technique [0002] With the acceleration of population aging, the sharp increase in patients with malignant tumors, AIDS, organ transplantation, and diabetes has led to the development of a large number of interventional diagnosis and treatment technologies and the use of broad-spectrum antibiotics and glucocorticoids. The current clinically acquired deep fungal infection is developing rapidly. and has become one of the leading causes of death in immunocompromised patients. Invasive pulmonary aspergillosis (IPA) caused by Aspergillus fumigatus is a common and fatal opportunistic infection in immunocompromised patients such as hematological malignancies, organ transplantation, and AIDS, with a mortality rate as h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/06A61K38/12A61K31/496A61K31/7048A61K31/506A61P31/10A61K31/11
Inventor 陈一强罗劲孔晋亮
Owner GUANGXI MEDICAL UNIVERSITY
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