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Medicine for restraining clostridium difficile thalli and/or restraining germination of clostridium difficile spores

A technology for Clostridium difficile and Clostridium difficile infection, which is applied in the field of medicines for inhibiting Clostridium difficile cells and/or inhibiting germination of Clostridium difficile spores

Pending Publication Date: 2019-11-22
CHENGDU BIOBEL BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN201710100322.7 discloses the preparation method and medical use of honokiol derivatives, but there is no report of such derivatives in the treatment of Clostridium difficile infectious diseases

Method used

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  • Medicine for restraining clostridium difficile thalli and/or restraining germination of clostridium difficile spores
  • Medicine for restraining clostridium difficile thalli and/or restraining germination of clostridium difficile spores
  • Medicine for restraining clostridium difficile thalli and/or restraining germination of clostridium difficile spores

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Magnolol and honokiol inhibit Clostridium difficile activity

[0026] Three strains of Clostridium difficile, ATCC BBA1870, ATCC 700057, and ATCC 630, were selected in the experiment, and were tested on Brussel agar medium with supplements. Strains frozen in glycerol at -80°C were inoculated on solid agar medium. Place in a 37°C incubator for anaerobic cultivation for 24 to 48 hours. The test range of magnolol and honokiol is 128 μg / ml-0.0625 μg / ml, and there are 11 two-fold dilution concentration gradients in total. Magnolol and honokiol were formulated into high-concentration working solutions with a concentration of 100 times respectively. They were used on the same day, and the solvent was 100% DMSO. For each dilution of the agar plate preparation, 20 μl of the high concentration working solution was mixed with 2 ml of molten, supplemented Brookfield agar (45-55° C.) and added to a six-well plate and allowed to solidify. 1% DMSO was used as a growth ...

Embodiment 2

[0030] The inhibition of embodiment 2 magnolol and honokiol to different anaerobic bacteria activity

[0031] Five strains of anaerobic bacteria, including ATCC 10031, ATCC 12022, ATCC 13048, ATCC 25611, and ATCC 25922, were selected in the experiment and tested on BHI agar medium with supplements. Strains frozen in glycerol at -80°C were inoculated on solid agar medium. Place in a 37°C incubator for anaerobic cultivation for 24 to 48 hours. The test range of magnolol and honokiol is 128 μg / ml-0.0625 μg / ml, and there are 11 two-fold dilution concentration gradients in total. Magnolol and honokiol were formulated into high-concentration working solutions with a concentration of 100 times respectively. They were used on the same day, and the solvent was 100% DMSO. For each dilution of the agar plate preparation, 20 μl of the high concentration working solution was mixed with 2 ml of molten, supplemented Brookfield agar (45-55° C.) and added to a six-well plate and allowed to s...

Embodiment 3

[0036] Example 3 Magnolol and honokiol inhibit Clostridium difficile spore germination experiment

[0037]According to the method disclosed in the literature The Journal of Infectious Diseases 2013; 207:1498-504, the spores of ATCC BBA1870, ATCC 43255, and ATCC 630 strains were cultivated and purified. Purified C. difficile spores were pelleted and washed three times with deionized water, and the buffer in which the spores were stored was removed by centrifugation at 9400 g. The spores were heat activated at 68 °C for 30 min and then washed three more times to remove any autogerminated spores. The turbidity of the spore liquid was adjusted to OD580=1.0 with 100 mM sodium phosphate buffer (pH6.0) containing 5 mg / mL sodium bicarbonate. The magnolol and honokiol were mixed with 6mM taurocholate and 12mM glycine, respectively, and added to 96-well plates. After adding the spores, measure the OD580 (counted as OD580(t)) every minute for 2 hours, and compare it with the OD580 at z...

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Abstract

The invention provides an application of a compound as shown in the formula A to preparation of a medicine for restraining clostridium difficile thalli and / or restraining germination of clostridium difficile spores. R1 and R2 are independently selected from C1-3 alkyl group, C2-3 alkenyl and C2-3 alkynyl. Experiment proves that the compound as shown in the formula A, particularly magnolol and honokiol can notably restrain activity of clostridium difficile, can notably restrain germination of clostridium difficile spores, and has good application prospects for preparation of the medicine for preventing and / or treating clostridium difficile infection diseases, recurrence of clostridium difficile infection diseases, and complications of clostridium difficile infection diseases.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a medicine for inhibiting Clostridium difficile thallus and / or germination of Clostridium difficile spores. Background technique [0002] Clostridium difficile (Clostridium difficile) is an obligate anaerobic bacterium of the genus Clostridium, which is very sensitive to oxygen and difficult to isolate and culture, hence the name, and generally parasitizes in the human intestinal tract. Usually Clostridium difficile infection is caused by excessive use of certain antibiotics, which breaks the balance of intestinal flora, makes the growth of Clostridium difficile flora accelerate, and causes inflammation. Clostridium difficile produces exotoxins A and B with different effects at different times. Toxin A is an enterotoxin, which first binds to mucosal cells in the early stage, causing primary damage, leading to inflammation of the intestinal wall, cell infiltration, i...

Claims

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Application Information

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IPC IPC(8): A61K31/05A61P1/12A61P1/00A61P31/04
CPCA61K31/05A61P1/12A61P1/00A61P31/04
Inventor 广兵阳泰董韧涵刘进谢建黄胜彭向阳覃传军赖永新占伟王晓辉
Owner CHENGDU BIOBEL BIOTECHNOLOGY CO LTD
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