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New application of locust endophytic fungus secondary metabolite

A technology of secondary metabolites and endophytic fungi, applied to medical preparations containing active ingredients, drug combinations, blood diseases, etc., can solve problems such as concerns and increased investment

Inactive Publication Date: 2021-11-05
CHINA THREE GORGES UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the large market opportunities and the shortcomings of existing drugs, pharmaceutical companies have significantly increased their attention and investment in the research and development of anticoagulant drugs

Method used

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  • New application of locust endophytic fungus secondary metabolite
  • New application of locust endophytic fungus secondary metabolite
  • New application of locust endophytic fungus secondary metabolite

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Inoculate the bacteria activated by PDA medium into PDB medium, add the precursor L-tryptophan with a final concentration of 1.0mM in batches at 28°C and 180rpm, and collect the fermentation broth and bacteria after continuing to cultivate for 20 days. Mycelia, extract the fermentation broth 3 times with ethyl acetate, dry the mycelia at 45°C, add acetone to soak overnight, extract by ultrasonic (45°C, 60W) for 30min, repeat the extraction 3 times, and combine all extracts The weight is 15g. The total extract was separated by normal phase silica gel (200-300 mesh) column chromatography, and chloroform-methanol=20:1 (volume ratio) transverse gradient elution was used to obtain 50 fractions in total. Carry out TLC detection, developing system is: chloroform-methanol=15: 1 (volume ratio), observe fluorescence under 365nm ultraviolet lamp, then spray color development with improved potassium iodide, observe whether there are orange spots, find that the first From the 25th ...

Embodiment 2

[0017] The anticoagulant activity determination of 5 compounds obtained according to embodiment 1

[0018] In order to evaluate the anticoagulant activity of Chuanxiongdole and its analogues, the direct thrombin inhibitor chromogenic assay was used to screen and compare. Direct thrombin inhibitor chromogenic assay: take hirudin as the positive drug, use bovine serum albumin buffer according to the kit operation, configure hirudin and compound solutions with the same concentration gradient, set up the experimental group and the blank group, and the experimental group Add 50 μl of prepared sample solution and 50 μl of thrombin substrate into a microwell plate at 37°C (the blank group does not add 50 μl of sample solution), mix well and incubate at 37°C for 2 minutes, then add 50 μl of thrombin and mix well Incubate at 37°C for 2 minutes, finally add 100 μl of 2% citric acid as a reaction stop solution, mix well, measure the absorbance (OD value) at 405 nm, perform three parallel...

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Abstract

The invention discloses an application of five compounds derived from an endophytic fungus Paracoiothyrium brasiliense in the aspect of anticoagulation, Four compounds of the five compounds have a remarkable inhibition effect on anticoagulant activity, the median inhibitory concentrations of the four compounds are respectively 7.11 [mu] g / ml, 13.11 [mu] g / ml, 13.59 [mu] g / ml and 2.23 [mu] g / ml, and the four compounds are obviously superior to positive drug hirudin, and can be used for being developed into a treatment coagulant drug. Therefore, the four compounds provided by the invention have clinical application potential of anticoagulant activity.

Description

technical field [0001] The invention relates to the application of anticoagulant activity of five compounds derived from the endophytic fungus Paraconiothyrium brasiliense, and belongs to the technical field of biopharmaceuticals. Background technique [0002] The prevention and treatment of thromboembolic diseases has been a research hotspot in recent years. Thrombosis, the formation of localized blood clots, is a major cause of development and death in arterial diseases such as myocardial infarction and stroke, as well as in venous thromboembolic diseases. Drug therapy plays an important role in the prevention and treatment process [Wang Xiu, Xia Quan. Current status and research progress of clinical application of anticoagulant drugs. Anhui Medicine, 2011,15(10):1189-1192.]. Anticoagulant therapy is the basis for the treatment of thromboembolic diseases. Its principle is to prevent the blood coagulation process by affecting coagulation factors. It is mainly used in the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/403A61K31/437A61P7/02
CPCA61K31/403A61K31/437A61P7/02
Inventor 刘呈雄李慧周继慧陈莲刘朝霞邹坤
Owner CHINA THREE GORGES UNIV