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Compounds isolated from Colletosporum anthracis and preparation method and use thereof

A technology of glenospora anthracnose and compounds, applied in biochemical equipment and methods, organic chemical methods, and methods based on microorganisms, to achieve the effects of easy control of preparation conditions, improvement of learning and memory ability, and simple preparation methods

Active Publication Date: 2020-06-05
LANZHOU UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there have been no reports on the isolation of glenospora anthraxin A from glenospora anthracnose and its inhibitory activity against acetylcholinesterase

Method used

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  • Compounds isolated from Colletosporum anthracis and preparation method and use thereof
  • Compounds isolated from Colletosporum anthracis and preparation method and use thereof
  • Compounds isolated from Colletosporum anthracis and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Preparation of compounds of the present invention:

[0036] (1) Fermentation of Glosporum anthracnose

[0037] a. Strain activation: take the frozen strain of G. anthracis, inoculate it on the plate of PDA solid medium in the ultra-clean workbench, and cultivate it in a constant temperature incubator at 28°C for 7 days;

[0038] b. Inoculation and primary fermentation of strains: Inoculate the activated strains into 500ml Erlenmeyer flasks containing 200mL PDB liquid medium, a total of 12 flasks were placed in a 28°C constant temperature air bath rotary shaker, shaker The rotation speed is 160r / min, the culture time is 10 days, and the total fermentation volume is 2.4L;

[0039] c. Large-scale fermentation of strains: prepare a total of 45L of PDB liquid medium, sterilize it on a steam generator, the sterilization temperature is 115-118°C, and the sterilization time is 2.5h. After the sterilization is completed, cool the medium to room temperature Dissolve one bottle ...

Embodiment 2

[0047] Determination and verification of the structural formula of the compound of the present invention:

[0048] (1) Physical and chemical property data of the compound

[0049] Colorless oil, optical rotation: 30; development system is petroleum ether: acetone = 2:1, the color of ethanol sulfate is light yellow spots.

[0050] (2) Determination of the molecular formula of the compound

[0051] combine 1 H-NMR and 13 C-NMR data and HRESIMS (measured value 273.1466[M+Na] + , calculate C 15 h 22 o 3 The Na value is 273.1461), and its molecular formula is determined to be C 15 h 22 o 3 .

[0052] (3) Determination of the compound structural formula

[0053] Such as figure 2 As shown, the IR spectrum data show that the compound contains a hydroxyl group and a carbonyl group (νmax=3406, 1724cm -1 ). 1 H-NMR, 13 C-NMR and DEPT spectra (such as Figure 3-Figure 5 Shown) shows three methyl groups, three sp3 methylene groups, one sp2 methylene group, two sp3 methine ...

Embodiment 3

[0064] The acetylcholinesterase inhibitory activity test of compound of the present invention:

[0065] (1) Determination of the acetylcholinesterase inhibitory activity of the compound of the present invention

[0066] Adopt improved Ellman method to measure the acetylcholinesterase inhibitory activity of compound gliosporine anthraxin A, huperzine A is used as positive control drug, concrete implementation steps are as follows:

[0067] The improved Ellman method was used for determination, and the operation steps were as follows: 140 μL PBS (0.1M pH=8.0), 20 μL sample solution (final concentration: 1 mg / mL), 15 μL AChE (0.28 U / mL, pH = 8.0 PBS dissolved dilution). After incubation at 4°C for 20 min, 10 μL DTNB (0.075 mol / L) and 10 μL ATCI (0.01 mol / L) were added. Incubate at 37°C for 20 min, and measure the absorbance at 405 nm with a microplate reader. Among them, 20 μL of PBS (pH 8.0) was used to replace 20 μL of sample solution in the blank group; 20 μL of huperzine A...

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Abstract

The invention discloses a compound separated from colletotrichum gloesporioides and a preparation method and application thereof. The molecular formula of the compound separated from the colletotrichum gloesporioides is C15H22O3. The preparation method of the compound comprises the following steps: activating a colletotrichum gloesporioides strain which has been cryopreserved, inoculating the activated strain to a culture medium for culturing, preparing a PDB liquid culture medium, and sterilizing; dissolving a bottle of penicillin and a bottle of streptomycin in sterile water, and adding the solution in a fermenting tank; inoculating the sterilized culture into the fermenting tank for fermenting; after fermenting is finished, carrying out suction filtration on a fermented product to obtain mycelium; extracting and concentrating to obtain a bacteria solution extract; and carrying out column chromatography, elution and wet packing on a coarse extract, carrying out elution and column chromatography on silica gel again, carrying out elution with petroleum ether-acetone eluant again, carrying out column chromatography, carrying out elution with chloroform-methanol to implement decolorization, and carrying out separation to obtain the compound which is colletotrichum gloesporioides A. The provided compound can be used for preparing anti-senile dementia drug.

Description

technical field [0001] The invention belongs to the technical field of medicine separation and preparation, and in particular relates to a compound isolated from glyospora anthracnose and its preparation method and application. Background technique [0002] Alzheimer's disease (Alzheimer's Disease, AD), that is, senile dementia, is a progressive, neurodegenerative disease, accompanied by memory loss and cognitive impairment. The number of patients with Alzheimer's disease has increased dramatically, and it has gradually become a social problem. The treatment of Alzheimer's disease has also become an urgent problem to be solved by the scientific community. [0003] The etiology and pathogenesis of Alzheimer's disease have not yet been elucidated. At present, domestic and foreign clinical studies have shown that acetylcholinesterase inhibitors are the most effective drugs for the treatment of AD, because drugs that inhibit acetylcholinesterase can significantly increase the l...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C49/743C07C45/78C07C45/79A61P25/28C12P7/26C12R1/645
CPCC07B2200/07C07C45/78C07C45/79C07C49/743C12P7/26
Inventor 杨中铎陈效威
Owner LANZHOU UNIVERSITY OF TECHNOLOGY
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