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Applications of daphnetin in inhibition of Helicobacter pylori

A technology of Helicobacter pylori and daphnetin, applied in the field of daphnetin

Active Publication Date: 2018-08-10
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on its activity inhibition against Helicobacter pylori (Hp)

Method used

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  • Applications of daphnetin in inhibition of Helicobacter pylori
  • Applications of daphnetin in inhibition of Helicobacter pylori
  • Applications of daphnetin in inhibition of Helicobacter pylori

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Amoxicillin critical plate: Dissolve Mueller Hinton Agar medium in double distilled water, 121°C, 15min. When it cools down to 55°C, add 5% sheep blood, mix well, then take 13.5ml and pour it into the mixture containing 5mg / ml, 10mg / ml, 20mg / ml amoxicillin China Institute for Food and Drug Control, production batch number 130409- 200208), dry it for later use. Example 1 The acquisition of multidrug-resistant Helicobacter pylori strains

[0032] 1. Culture and subculture of Helicobacter pylori

[0033] The collected Helicobacter pylori strains were cultured in Karmali blood plates, and the culture conditions were 10% CO 2 , 5%O 2 and 85%N 2 , and cultured at 37°C for 3 days. When Helicobacter pylori grows, take it out for passage, take out the Helicobacter pylori to be passaged from the micro-aerobic tank, put it into the ultra-clean workbench, scrape enough Helicobacter pylori colonies into the normal saline with the inoculation loop, and adjust the concentration t...

Embodiment 2

[0037] The determination of embodiment 2 clinical strain drug resistance phenotype

[0038] 1. Critical value method:

[0039] (1) Inoculation: Use an inoculation loop to scrape Hp grown in the resistant plate into normal saline, and dilute to McF2.0-4.0 (about 1×10 8 CFU / ml). Use a pipette gun to add the diluted bacterial solution (2.5 μL / spot) into critical plates containing different types and concentrations of antibiotics.

[0040] The types of antibiotics are specifically metronidazole MTZ, clarithromycin CLA, levofloxacin LEV, amoxicillin AMX, and tetracycline TE.

[0041] (2) Cultivation: Put the plate into the culture tank and put it into the gas bag so that the gas condition is 10% CO 2 , 5%O 2 and 85%N 2 , and cultivated at 37°C for 72h.

[0042] (3) Result reading:

[0043] If bacteria grow in the critical plate, it is recorded as the drug-resistant bacteria of the corresponding antibiotic.

[0044] If the bacteria grow below the critical value but the critica...

Embodiment 3

[0057] Embodiment 3 drug susceptibility test

[0058] 1. Paper method

[0059] (1) Coating: Use an inoculation loop to scrape Hp grown in the resistant plate into normal saline, and dilute to McF2.0-4.0 (about 1×10 8 CFU / ml). Use a pipette gun to spread 150 μL of the diluted bacterial solution on an MH blood plate to dry, and then place a drug-sensitive paper sheet (oxioid, production batch number) containing 100 μg daphnetin (Dalian Meilun Biotechnology Co., Ltd., production batch number A1228AS) 1829617) pasted in.

[0060] (2) Cultivation: Put the plate into the culture tank and put it into the gas bag so that the gas condition is 10% CO 2 , 5%O 2 and 85%N 2 , and cultivated at 37°C for 72h.

[0061] (3) Result reading: the diameter of the inhibition zone was measured by a vernier caliper.

[0062] 2. Plate two-fold dilution method

[0063] (1) Inoculation: Use an inoculation loop to scrape Hp grown in the resistant plate into normal saline, and dilute to McF 2.0-4....

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Abstract

The present invention relates to new uses of daphnetin in inhibition of Helicobacter pylori. According to the present invention, the daphnetin can well kill a variety of drug-resistant phenotype Helicobacter pylori; particularly the results show that the daphnetin can produce the zone-of-inhibition with a diameter of 39-53 mm at a concentration of 100 [mu]g / disc, and has the minimum inhibitory concentration (MIC) of 25-100 [mu]g / ml; and the new use of the daphnetin provide the new option for Helicobacter pylori inhibition drugs.

Description

technical field [0001] The invention relates to a new application of daphnetin, in particular to its application in anti-helicobacter pylori, and belongs to the field of medical biotechnology. Background technique [0002] Daphnetin (Daphnetin) is an active ingredient extracted from Daphne Korean Nakai, also known as Zushimajialin. It is the first new drug in my country, and its chemical name is 7,8-dihydroxycoumarin (7,8-dithydroxycoumarin), mainly found in the genus Daphneaceae. [0003] Daphnetin has a wide range of pharmacological effects, including: antithrombotic, hypolipidemic, anti-inflammatory, analgesic, sedative and hypnotic, antimalarial, aphidicidal, and antibacterial effects. At present, its antibacterial activity has been reported, mainly including: Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and so on. However, there is no report on its activity inhibition against Helicobacter pylori (Hp). Contents of the inventio...

Claims

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Application Information

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IPC IPC(8): A61K31/37A61P31/04A61P1/00A61P1/04A61P35/00
CPCA61K31/37A61P1/00A61P1/04A61P31/04A61P35/00
Inventor 游雪甫王根柱李聪然杨信怡王秀坤熊雁琼庞晶卢曦胡辛欣聂彤影李雪卢芸
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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