Application of naphthoγ-pyrone compounds in the preparation of anti-Helicobacter pylori drugs or preventive health care products

A technology of pyrone and compound, which is applied in the field of marine fungal metabolites as new antibacterial agents, and achieves the effects of simple operation, high extraction yield and high product purity

Active Publication Date: 2022-07-19
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no reports have been retrieved so far that the naphtho-γ-pyrone compounds derived from Aspergillus niger have anti-Helicobacter pylori activity.

Method used

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  • Application of naphthoγ-pyrone compounds in the preparation of anti-Helicobacter pylori drugs or preventive health care products
  • Application of naphthoγ-pyrone compounds in the preparation of anti-Helicobacter pylori drugs or preventive health care products
  • Application of naphthoγ-pyrone compounds in the preparation of anti-Helicobacter pylori drugs or preventive health care products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Isolation of fungal DM94

[0042] The strain DM94 was isolated from the root soil of the mangrove tree. The soil samples were first placed in a sterile petri dish and air-dried in the natural environment. Then, 1 g of the air-dried soil samples were weighed and dissolved in 10 mL of 50% old seawater to obtain 10 mL of 50% old seawater. -1 solution, continue to dilute to 10 -2 , 10 -3 , to obtain three gradient sample solutions. Take 100 μL respectively and spread it on the separation medium GPY medium, Martin medium, CA medium, PDA medium, SDA medium and fungus No. 2 medium. Two separate mediums with different dilution concentrations are in parallel. Culture at 28°C for 1 to 8 weeks. The single colonies were observed and picked, and were inoculated on the fungus No. 2 medium to obtain pure cultured strains.

Embodiment 2

[0043] Example 2 Molecular identification of fungal DM94

[0044] Extraction of genomic DNA: Obtain the DM94 genomic DNA template using a nucleic acid rapid extractor. The operation is as follows: Take a sterile DNA extraction tube filled with quartz sand, add 500 μL of TE, pick an appropriate amount of mycelium into the tube, and extract by high-speed shaking 1min. Add 500 μL of phenol-chloroform solution and centrifuge to obtain genomic DNA.

[0045] Amplification of gene sequence: 12.5μL of 2x PCR mastermix was added, primers ITS1 (5'-TCC GTAGGT GAA CCT GCG G-3') and ITS4 (5'-TCC TCC GCT TAT TGA TAT GC-3'), ( 18SF is 5'-AACCTG GTT GAT CCT GCC AGT-3'; 18SR is 5'-CGA CGG GCG GTG TGT AC-3') 1 μL each, template DNA 2 μL, and double distilled water 13.5 μL. The amplification conditions were pre-denaturation at 95°C for 3 min; denaturation at 95°C for 30S, annealing temperature at 59°C for 30S, extension temperature at 72°C for 30S, a total of 32 cycles; post-extension at 72°C ...

Embodiment 3

[0049] The small preparation of the fermentation crude extract of embodiment 3 fungus DM94

[0050] Bacteria were grown in corn solid fermentation medium (corn kernels 50g; ammonium tartrate 2.37g; yeast extract 0.86g; KH 2 PO 4 0.25g; MgSO 4 0.17g; sea salt 0.4g; distilled water 20mL, natural pH. (Hong Kui, Yang Xue, A high-yielding pseudoshell compound strain Aspergillus pyroli TKYX429 and its application, ZL201310186483.4)) 28° C., cultured for 4 weeks. First soak the fermentation product with 80% acetone for 12h, then ultrasonicate for 20min with an ultrasonic extractor, extract with ethyl acetate for 3 to 5 times, combine the extracts, evaporate to dryness, and finally dissolve in chromatographic methanol. The samples were used for antibacterial activity detection and HPLC detection.

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Abstract

The invention discloses the application of a class of naphtho-γ-pyrone compounds in the preparation of anti-Helicobacter pylori medicines or preventive health care products. The present invention provides a marine fungus named Aspergillus sp.DM94, preservation number: CCTCC NO: M 20191003. The invention utilizes the polarity difference of naphtho-γ-pyrones to extract and separate a class of naphtho-γ-pyrone compounds from the fermentation culture of marine fungi, and the method is easy to operate, high in extraction yield and high in product purity , suitable for large-scale production. The in vitro antibacterial experiments show that the naphtho γ-pyrones have good in vitro antibacterial activity, and the MIC value is against the standard strain G27 of Helicobacter pylori and the multidrug-resistant strain HP159 of levofloxacin, clarithromycin and metronidazole. 4~64μg / mL, can be used to prepare antibacterial drugs and preventive health food, and has a good development prospect.

Description

technical field [0001] The invention relates to the field of marine fungal metabolites as new antibacterial agents, in particular to the application of a class of naphthoγ-pyrone compounds in the preparation of anti-Helicobacter pylori medicines or preventive health care products. Background technique [0002] Helicobacter pylori is a microanaerobic, Gram-negative bacterium that is often located in the fundus mucosa of the human stomach and infects about half of the world's population. Helicobacter pylori infection often leads to digestive diseases such as chronic gastritis and peptic ulcers. Studies have shown that Helicobacter pylori infection is closely related to the occurrence and development of gastric cancer, and eradication of Helicobacter pylori can reduce the risk of gastric cancer. For the clinical treatment of Helicobacter pylori, triple or quadruple therapy recommended by relevant guidelines, including the proton pump inhibitor omeprazole and the antibiotics am...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P17/06C12P17/16A61K31/352A61P31/04A23L33/10A23L31/00C12R1/66
CPCC12N1/14C12P17/06C12P17/162A61K31/352A61P31/04A23L33/10A23L31/00A23V2002/00C12R2001/66A23V2200/30A23V2250/208Y02A50/30
Inventor 唐金山洪葵毕洪凯姚新生董治统贾佳苟小霜薛雅馨丁文娟
Owner JINAN UNIVERSITY
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