CN115252594A

CN115252594A - Application of sanshool in preparation of medicine for resisting propionibacterium acnes

Info

Publication number: CN115252594A
Application number: CN202211021663.2A
Authority: CN
Inventors: 蒋献; 周夕湲
Original assignee: West China Hospital of Sichuan University
Current assignee: West China Hospital of Sichuan University
Priority date: 2021-09-27
Filing date: 2022-08-24
Publication date: 2022-11-01

Abstract

The invention provides application of sanshool in preparing a medicine for resisting propionibacterium acnes. Also discloses a medicine for resisting propionibacterium acnes, which is prepared by taking sanshool as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients. The sanshool can effectively resist propionibacterium acnes, relieve inflammatory reaction caused by the propionibacterium acnes, and provide a new choice for clinically treating diseases caused by the propionibacterium acnes, such as acne, folliculitis and the like.

Description

Application of sanshool in preparation of medicine for resisting propionibacterium acnes

Technical Field

The invention belongs to the field of biological medicines, and particularly relates to application of sanshool in preparation of a medicine for resisting propionibacterium acnes.

Background

Propionibacterium acnes (p.acens) is a gram-positive anaerobic or facultative anaerobic bacterium widely distributed in the skin, hair, oropharynx and gastrointestinal tract of the human body, and its pathogenic effects are mainly to make lipids in sebum form long-chain fatty acids, stimulate the local part and cause obstruction of the sebum tube, which is one of the main factors causing acne. In 1951, andrews et al take the lead to oral tetracycline treatment of acne with good therapeutic effect, so that the treatment of acne by oral administration of antibiotics to suppress Propionibacterium acnes is considered to be the safest and efficient new way to treat acne. To date, dermatologists have also widely used oral antibiotics for the treatment of inflammatory acne. However, oral administration of antibiotics often causes various adverse effects such as gastrointestinal reactions and the like, and long-term use of the antibiotics causes drug resistance of propionibacterium acnes to the antibiotics, so that the clinical curative effect is greatly reduced. Therefore, there is a need to find new drugs capable of effectively inhibiting and killing propionibacterium acnes, so as to be used for the clinical treatment of acne.

The pricklyash peel is the peel of Rutaceae (Rutaceae) Zanthoxylum plant (Zanthoxylum), and the recorded effects of the Chinese medicinal materials include warming spleen and stomach for dispelling cold, eliminating dampness, relieving pain, killing parasite, treating food retention and fluid retention, psychroalgia of heart and abdomen, toothache, pudendal pruritus, scabies and sores. The amide substances are the most characteristic alkaloid in the pepper, are the numb taste sources of the pepper, are separated and identified for more than 50 at present, and are represented by sanshool. In recent years, the sanshool has been found to have a wide range of biological effects, including promoting gastric motility, anaesthetizing and easing pain, expelling parasites, relieving itching, resisting tumors, resisting platelet aggregation and the like. However, the effect of sanshool against propionibacterium acnes has not been reported at present.

Disclosure of Invention

The invention aims to provide a new application of sanshool.

The invention provides application of sanshool in preparing a medicine for resisting propionibacterium acnes.

Further, the anti-propionibacterium acnes drug is a drug for killing or killing propionibacterium acnes.

Furthermore, the anti-propionibacterium acnes drug is a drug for killing or killing propionibacterium acnes. Furthermore, the anti-propionibacterium acnes drug is a drug for inhibiting the growth of propionibacterium acnes.

Furthermore, the medicine is a medicine for treating diseases caused by propionibacterium acnes.

Further, the above-mentioned medicament is a medicament for treating acne and/or folliculitis.

Further, the sanshool is extracted from zanthoxylum bungeanum maxim plants.

Further, the sanshool is one or more of hydroxy- α -sanshool, hydroxy- β -sanshool, hydroxy- γ -sanshool, hydroxy- δ -sanshool, hydroxy-e-sanshool, α -sanshool, β -sanshool, γ -sanshool, δ -sanshool, and e-sanshool.

Preferably, the sanshool is α -sanshool.

Further, the purity of the sanshool is higher than 95%, preferably not lower than 98%, more preferably higher than 98%.

The invention also provides a medicine for resisting propionibacterium acnes, which is a preparation prepared by taking sanshool as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients. Preferably, the preparation is a liquid preparation, wherein the content of sanshool is not less than 1.25mg/mL; more preferably, the adjuvant of the liquid preparation is ethanol.

Further, the above preparation is an external preparation.

The invention proves that the sanshool has the beneficial effects of resisting the propionibacterium acnes and relieving inflammation caused by the propionibacterium acnes, and provides a new choice for clinically treating diseases caused by the propionibacterium acnes, such as acne, folliculitis and the like.

It will be apparent that various other modifications, substitutions and alterations can be made in the present invention without departing from the basic technical concept of the invention as described above, according to the common technical knowledge and common practice in the field.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1 shows the results of a droplet plate experiment of sanshool against Propionibacterium acnes.

FIG. 2 is the statistical results of the droplet plate experiment of sanshool against Propionibacterium acnes.

FIG. 3 is the results of treatment with sanshool on a murine ear model of Propionibacterium acnes.

Detailed Description

Unless otherwise stated. The raw materials used in the invention are all known products and are obtained by purchasing commercial products.

Example 1 bacteriostatic Effect of hydroxy-alpha-Zanthoxylum piperitum on Propionibacterium acnes in vitro

1. Determination of Minimum Inhibitory Concentration (MIC) by 96-well plate brain heart infusion Broth (BHI) method

Experimental materials: selecting a propionibacterium acnes (c.acne) standard strain: ATCC 6919 (deposited center for species, institute of microbiology, guangdong province) and hydroxy- α -sanshool (HAS, mcSt, inc., RM1978, purity 98%), brain heart infusion Broth (BHI) (Haibo, HB8297-5, concentration 98%).

Preparing HAS mother liquor: 31.6mg HAS was made up with 316ul absolute ethanol to a stock solution concentration of 100 mg/ml.

The propionibacterium acnes cultured to the peak of the growth curve is proportioned into a bacterial suspension with OD60=0.3 (the bacterial density is 0.3 multiplied by 10) ⁸ One/ml); 200ul of sterile PBS was added to one ring of wells around the 96-well plate as a barrier, 200ul of bacterial suspension was added to the first well, and 100ul of the rest was added to each wellul bacteria suspension. Then 50ul of HAS stock was added from the first well, then 100ul was aspirated in sequence, so the sequential concentrations were: 5mg/ml,2.5mg/ml,1.25mg/ml,0.625mg/ml,0.313mg/ml,0.156mg/ml,0.078mg/ml,0.039mg/ml,0.019mg/ml, blank control (bacterial suspension only); putting the 96-well plate into an anaerobic incubator, adding an anaerobic culture bag and an anaerobic indicator, and incubating for 3 days in the incubator at 37 ℃.

As a result: MIC of HAS to propionibacterium acnes =1.25mg/ml

2. Drop plate experiments

Experimental materials: the Propionibacterium acnes standard strain and the hydroxy-alpha-sanshool were the same as before. Agar powder, columbia CNM agar basic culture medium, a defibering poplar and snow 10cm culture dish, a constant temperature incubator, anaerobic culture medium, an anaerobic air bag and an anaerobic indicator.

Preparing HAS mother solution: HAS stock solutions were prepared in DMSO at a concentration of 200 mg/ml.

The experimental steps are as follows: heating and melting the CNM culture medium, and heating and melting 1% agar solid; and when the temperature of the CNM culture medium is reduced to about 40 ℃, adding 5% defibrinated goat blood, uniformly mixing, pouring the mixture into a flat plate until the mixture is completely solidified, and preparing the blood flat plate culture medium. Adding HAS mother liquor into 500ul CNM culture medium at a certain proportion, adding 2.5ul mother liquor per 100ul, respectively preparing 1.25mg/ml,2.5mg/ml,5mg/ml, and preparing 1.16 × 10 ⁸ The bacterial solution/ml was aspirated out 100ul,1.2 million rotations, centrifuged for 2 minutes, the original culture medium was aspirated, and medium prepared with HAS of various concentrations was added. Different concentrations of HAS and Propionibacterium acnes were allowed to act for 2h, 4h, 6h, and 8h, respectively, and then dropped onto blood plates at 1 point for each 10-fold drop in bacterial suspension concentration for a total of 5 points, with 3 replicates per plate (see FIG. 1). Finally, calculating the number of the last colony with clear energy, and measuring and calculating the in-vitro bacteriostatic action of the HAS.

The results are shown in FIG. 2, further reflecting that HAS HAS a bacteriostatic effect on Propionibacterium acnes, with a MIC of 1.25mg/ml.

EXAMPLE 2 therapeutic Effect of hydroxy-alpha-Zanthoxylin on Propionibacterium acnes infected murine ear model

1. Experimental materials: balb/c Male mice (6-8W), selecting a Propionibacterium acnes standard strain: ATCC 6919 (Collection of species, institute of microbiology, guangdong province) and hydroxy- α -sanshool (McBiotech, inc., RM 1978), brain heart leach Broth (BHI) (Haibo, HB8297-5, 98% concentration).

The proportion of the bacterial suspension is as follows: the Propionibacterium acnes in the log phase of growth is washed twice with sterile PBS, and then mixed with sterile normal saline to obtain a mixture of 1 × 10 ⁸ Bacterial solution/ml. HAS was prepared in 20mg/ml absolute ethanol.

The experimental contents are as follows: 24 mice were randomly divided into 3 groups: model group (left ear root injected with 25ul of bacterial suspension, right side injected with 25ul of physiological saline), HAS group (left ear root injected with 25ul of bacterial suspension, immediately coated with 75ul of HAS solution, 12h later coated with the same dosage of HAS solution), and stroma group (left ear root injected with 25ul of bacterial suspension, immediately coated with 75ul of absolute ethanol solution, 12h later coated with the same dosage of absolute ethanol solution). Mice were sacrificed 24 hours later, photographed, local harvested and fixed in formaldehyde.

The experimental results are as follows: the left mouse ear of the HAS group was red and swollen, and had a significantly reduced thickness compared to the absolute ethanol and the model group, and HE stained left ear with reduced thickness and inflammatory response (see fig. 3).

The experimental results prove that the sanshool has remarkable anti-propionibacterium acnes and anti-inflammatory effects, and has potential application value in treating acne.

In conclusion, the invention proves that the sanshool has the beneficial effects of resisting the propionibacterium acnes and relieving inflammatory reaction caused by the propionibacterium acnes, and provides a new choice for clinically treating diseases caused by the propionibacterium acnes, such as acne, folliculitis and the like.

Claims

1. Application of sanshool in preparing anti-Propionibacterium acnes medicine is provided.

2. The use of claim 1, wherein the anti-Propionibacterium acnes medicament is a medicament that kills or kills Propionibacterium acnes.

3. The use of claim 1, wherein the anti-propionibacterium acnes medicament is a medicament that inhibits the growth of propionibacterium acnes.

4. The use of claim 1, wherein the medicament is a medicament for treating a disease caused by propionibacterium acnes.

5. Use according to claim 4, wherein the medicament is a medicament for the treatment of acne and/or folliculitis.

6. The use according to claim 1, wherein the sanshool is one or more of hydroxy- α -sanshool, hydroxy- β -sanshool, hydroxy- γ -sanshool, hydroxy- δ -sanshool, hydroxy-e-sanshool, α -sanshool, β -sanshool, γ -sanshool, δ -sanshool, and e-sanshool.

7. The use according to claim 6, wherein the sanshool is hydroxy- α -sanshool.

8. The use according to any one of claims 1 to 7, wherein the sanshool is more than 95% pure; preferably, the purity of the sanshool is not less than 98%.

9. A medicine for resisting propionibacterium acnes is characterized by being a preparation prepared by taking sanshool as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients; preferably, the preparation is a liquid preparation, wherein the concentration of the sanshool is not lower than 1.25mg/mL; more preferably, the adjuvant of the liquid preparation is ethanol.

10. The medicament of claim 9, wherein the formulation is an external formulation.