Compound separated from colletotrichum gloesporioides and preparation method and application thereof

A technology of anthracnose and compounds, which are applied in biochemical equipment and methods, organic chemistry methods, microorganism-based methods, etc., to achieve the effects of improving learning and memory ability, easily controllable preparation conditions, and simple preparation methods

Active Publication Date: 2017-07-21
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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  • Compound separated from colletotrichum gloesporioides and preparation method and application thereof
  • Compound separated from colletotrichum gloesporioides and preparation method and application thereof
  • Compound separated from colletotrichum gloesporioides and preparation method and application 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 rotary shaker in a constant temperature air bath at 28°C. 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) Physicochemical property data of the compound

[0049] Colorless oil, optical rotation: 30; the developing system is petroleum ether:acetone=2:1, and the color of sulfuric acid ethanol 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 (found 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 compound structural formula

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

Embodiment 3

[0064] Acetylcholinesterase inhibitory activity test of the compounds of the present invention:

[0065] (1) Measurement of acetylcholinesterase inhibitory activity of the compounds of the present invention

[0066] Adopt the modified Ellman method to measure the acetylcholinesterase inhibitory activity of the compound anthraxine A, and Huperzine A is used as the positive control drug, and the specific implementation steps are as follows:

[0067] The modified Ellman method was used for determination. 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, 15 μL AChE (0.28 U / mL, pH=8.0 PBS solubilization 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, the blank group used 20 μL PBS (pH 8.0) to replace 20 μL of sample solution...

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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 Applications(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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