Compound extracted from symbiotic fungus Paracoiothyrium brasiliense and application of compound

A technology for symbiotic fungi, compounds, applied in the separation/purification of carbonyl compounds, microorganisms, microorganism-based methods, etc., can solve problems such as drug resistance, high bleeding risk, and low cure rate

Pending Publication Date: 2022-04-29
CHINA THREE GORGES UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although aspirin, clopidogrel and tirofiban are representative anti-platelet aggregation clinical western medicines have a long history of use [Zhang Lijuan. Clinical effect of aspirin tablets combined with clopidogrel bisulfate tab...

Method used

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  • Compound extracted from symbiotic fungus Paracoiothyrium brasiliense and application of compound
  • Compound extracted from symbiotic fungus Paracoiothyrium brasiliense and application of compound
  • Compound extracted from symbiotic fungus Paracoiothyrium brasiliense and application of compound

Examples

Experimental program
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Effect test

Embodiment 1

[0040]A method for the preparation of four new compounds derived from a class of symbiotic fungi, the four new compounds are isolated from the fermentation broth of Paraconiothyrium brasiliense strains isolated from the intestines of the Chinese sword-horned locust in the Shennongjia area .

[0041] The Paraconiothyrium brasiliense strain was preserved in the China Center for Type Culture Collection (Wuhan University) on May 25, 2016, address: Luojia Mountain, Wuchang, Wuhan City, Hubei Province, and the preservation name is: Cyclospora brasiliensis MZ-1, The classification name is Paraconiothyrium brasiliense MZ-1, and the deposit number is CCTCC M2016279.

[0042] The concrete preparation method of described 4 compounds is as follows:

[0043] Insert the symbiotic fungus Paraconiothyrium brasiliense frozen into the PDA solid medium for recovery, cultivate for 3 days to obtain the seed bacterial block, select the seed bacterial block of the size of 0.5cm * 0.5cm to insert in...

Embodiment 2

[0050] Compound 1: Crystalline (ethyl acetate). By high resolution mass spectrometry HR-ESI-MS m / z:251.1646[M+H]+(C 15 h 23 o 3 , calcd.251.1647), determine its molecular formula as C 15 h 22 o 3 , with an unsaturation of 5. Compound 1 1 H-NMR spectrum (Table 1) shows 2 alkenes proton signal δ H 5.89 (1H, d, J=9.8Hz, H-2), 7.08 (1H, d, J=9.8Hz, H-3), 4 methylene proton signals δ H 1.98(1H, t, J=12.9Hz, H-6a), 2.74(1H, d, J=12.9Hz, H-6b), 1.51(1H, m, H-8a), 1.59(1H, m, H -8b), 1.17 (1H, dd, J=12.2, 3.7, Hz H-9a), 1.89 (1H, t, J=3.1Hz, H-9b), 3.22 (1H, d, J=10.8Hz, H -12a), 3.26 (1H, d, J=10.8Hz, H-12b), 3 methyl proton signals δ H 1.02 (3H, s, H-13), 1.13 (3H, s, H-14), 1.90 (3H, s, H-15). Compound 1 13 In the C-NMR spectrum (Table 1), HSQC spectrum and DEPT spectrum, there are 15 carbon signals, including a carbonyl carbon signal δ C 206.4(C-1); four olefinic carbon signals δ C 122.2(C-2), 148.9(C-3), 120.7(C-4) and 153.3(C-5); two oxycarbon signals δ C 73.4 (C-...

Embodiment 3

[0066] Anti-platelet aggregation activity of 4 compounds obtained according to Example 1

[0067] In order to evaluate the anti-platelet aggregation activity of the compounds, some compounds were screened and compared by direct anti-platelet aggregation inhibitor chromogenic assay. Direct anti-platelet aggregation inhibitor chromogenic assay method: with aspirin as positive drug, positive drug and compound are dissolved with 5% DMSO physiological saline solution, select appropriate concentration (1mmol / L) sample to measure, experiment is set up experimental group and blank group, Mix the fresh plasma with sodium citrate (9:1, V / V), centrifuge at 1500g for 10 minutes to obtain the supernatant, preheat it in a 37°C incubator, take 50 μL of the compound and 450 μL of the plasma supernatant and mix well, After reacting at 37° C. for 5 minutes, 20 μL of ADP (10 μmol / L) was added, and the OD value was measured at a wavelength of 405 . Three parallel operations were performed, and t...

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Abstract

The invention provides a compound extracted from a symbiotic fungus Paraciothyrium brasiliense and application of the compound. The symbiotic fungus Paraciothyrium brasiliense is inoculated into a suitable fermentation culture medium for fermentation culture, the fermentation culture medium is a PDA solid culture medium, a seed block is obtained, the seed block is inoculated into a PDB culture medium for culture, a seed solution is obtained, and the seed solution is inoculated into the PDB culture medium for shake cultivation. Performing separation and extraction on the fermentation product, filtering the fermentation product to obtain mycelium and fermentation liquor, and extracting and concentrating the fermentation liquor; and drying, extracting and concentrating the mycelium, combining the extracted products, and carrying out chromatography preparation and separation to obtain the compound. Wherein the compound 3 and the compound 4 have a certain degree of anti-platelet aggregation activity, and the two compounds can obviously improve the content of cAMP in mouse plasma.

Description

technical field [0001] The invention relates to four new compounds derived from the fungus Paraconiothyrium brasiliense, an extraction method thereof and application of anti-platelet aggregation activity, belonging to the technical field of biopharmaceuticals. Background technique [0002] Thrombosis is an important factor causing cardiovascular diseases, and the main reason for thrombus is the adhesion, activation and aggregation of platelets under abnormal conditions, among which the aggregation of platelets is the most common, and the inhibition of platelet aggregation is the main means of antithrombotic treatment [Patrono C, Morais J, Baigent C, et al. Antiplatelet agents for the treatment and prevention of coronary atherothrombosis. J. Am. Coll. Cardiol., 2017, 70(14): 1760-1776.]. At present, the discovery and development of new anti-platelet aggregation target drugs is a hot field, and finding reasonable and effective anti-platelet aggregation drugs has great research...

Claims

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Application Information

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IPC IPC(8): C07C49/733C07D311/76C07D209/18A61P7/02C07C45/80C07C45/79C12P7/26C12P17/06C12P17/10C12R1/645
CPCC07C49/733C07D311/76C07D209/18A61P7/02C07C45/80C07C45/79C12P7/26C12P17/06C12P17/10C07C2602/28C07B2200/13Y02A50/30
Inventor 刘呈雄李慧陈莲尉小琴周继慧刘朝霞邹坤
Owner CHINA THREE GORGES UNIV
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