阅读说明：本技术 一种抗菌组合物及其在制备表皮葡萄球菌群体感应抑制剂中的应用 (Antibacterial composition and application thereof in preparation of staphylococcus epidermidis quorum sensing inhibitor ) 是由 沈志滨 胡锡昌 张衍湖 ***榕 于 2020-05-15 设计创作，主要内容包括：本发明公开了一种抗菌组合物,包含梣酮、白鲜碱和黃柏酮；同时提供了该抗菌组合物在制备表皮葡萄球菌抑制剂中的应用、在制备治疗和/或预防表皮葡萄球菌过度增殖引起的相关皮肤软组织感染疾病的药物中的应用以及在制备菌群群体感应系统抑制剂中的应用。所述抗菌组合物对于表皮葡萄球菌尤其是耐药性的表皮葡萄球菌有良好的抑菌效果,其机理是通过抑制抑制自诱导寡肽类(AIP)介导的群体感应系统关键基因sarA和LuxS/AI-2介导的群体感应系统关键基因luxS的表达,从而抑制致病菌群群体感应系统,进而抑制菌体表面生物被膜形成。该抗菌组合物组分来源与天然中药植物,具有无毒、无刺激等优点。(The invention discloses an antibacterial composition, which comprises fraxinellone, dictamnine and thujone; simultaneously provides the application of the antibacterial composition in preparing an staphylococcus epidermidis inhibitor, the application in preparing a medicament for treating and/or preventing related skin soft tissue infection diseases caused by excessive proliferation of staphylococcus epidermidis and the application in preparing a flora quorum sensing system inhibitor. The antibacterial composition has a good antibacterial effect on staphylococcus epidermidis, particularly drug-resistant staphylococcus epidermidis, and the mechanism is that the expression of a quorum sensing system key gene sarA mediated by self-induced oligopeptide (AIP) and the expression of a quorum sensing system key gene luxS mediated by LuxS/AI-2 are inhibited, so that a quorum sensing system of pathogenic bacteria is inhibited, and further the formation of a bacterial surface biofilm is inhibited. The antibacterial composition has the advantages of no toxicity, no irritation, etc. due to its component sources similar to those of natural Chinese medicinal plants.)

1. An antibacterial composition is characterized by comprising fraxinellone, dictamnine and thujone.

2. The antimicrobial composition according to claim 1, characterized by comprising the following components in mass percent: 30-45% of fraxinellone, 25-35% of dictamnine and 25-35% of cypress ketone.

3. Use of the antibacterial composition according to claim 1 or 2 for the preparation of an inhibitor of staphylococcus epidermidis.

4. The use of claim 3, wherein the Staphylococcus epidermidis which the inhibitor of Staphylococcus epidermidis is capable of inhibiting is a drug-resistant Staphylococcus epidermidis.

5. Use of the antibacterial composition according to claim 1 or 2 for the preparation of a medicament for the treatment and/or prevention of associated skin soft tissue infection diseases caused by excessive proliferation of staphylococcus epidermidis.

6. The use of claim 5, wherein the associated skin soft tissue infection disease comprises at least one of a wound infection, an epidermal abscess, cellulitis, and a chronic wound ulcer.

7. Use of the antibacterial composition according to claim 1 or 2 for the preparation of an inhibitor of quorum sensing systems.

8. An inhibitor of staphylococcus epidermidis, comprising the antibacterial composition according to claim 1 or 2.

9. A medicament for treating and/or preventing diseases related to skin soft tissue infection caused by excessive proliferation of staphylococcus epidermidis, comprising the antibacterial composition according to claim 1 or 2.

10. An inhibitor of quorum sensing systems, comprising the antibacterial composition of claim 1 or 2.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an antibacterial composition and application thereof in preparation of a staphylococcus epidermidis quorum sensing inhibitor.

Background

The antibacterial drug is a drug with bactericidal or bacteriostatic activity, and comprises various chemically synthesized drugs such as antibiotics, sulfonamides, imidazoles, nitroimidazoles, quinolones and the like. The antibacterial drugs have the functions of inhibiting and killing pathogens under certain concentration, and the drug resistance of the antibiotics in clinical application gradually appears due to different antibacterial spectrums of different antibacterial drugs and along with the large-scale use of the antibiotics. In recent years, the eyes are gradually shifted to the combined application of antibiotics and quorum sensing inhibitors at home and abroad, and the quorum sensing inhibitors are applied to destroy the quorum sensing of flora, so that the mutual connection among the flora is inhibited, the generation of biofilm is inhibited, the dosage of the antibiotics is further reduced, the drug resistance of the antibiotics is reduced, and the clinical antibiotic antibacterial effect is improved. Therefore, the research and application of the quorum sensing inhibitor have important clinical application value.

Staphylococcus epidermidis (Staphylococcus epidermidis) is a gram-positive coccus which grows on the epidermis of a living body, exists in the skin, vagina and other parts of a human body, and is named as Staphylococcus epidermidis because the Staphylococcus epidermidis is often piled up in a string shape. The genus is a normal flora type, most of which are nonpathogenic bacteria, and the few of which can cause other diseases, and is an important source for cross infection in hospitals. The related skin soft tissue infection diseases caused by the excessive proliferation of staphylococcus epidermidis comprise wound infection, epidermal abscess, cellulitis, chronic wound ulcer and the like. At present, the problem of antibiotic resistance continues to be one of the important factors that plague antibacterial therapy, and resistant staphylococcus epidermidis infection may lead to more serious diseases and consequences.

The Quorum sensing system (QS) is a biological system which promotes the production of microbial cell self-induced signal molecules (AI) through population density, and initiates specific gene expression through AI induction or inhibition biological signal transmission to achieve the intracellular and intercellular information communication of microbes. The quorum sensing system is widely existed in microorganisms such as bacteria, fungi and the like, regulates and controls various physiological functions, wherein the quorum sensing system comprises extracellular polysaccharide, effector protein, spore generation, biofilm formation, virulence factors and the like, can effectively improve the stress resistance and environmental adaptability of microbial populations, and is also one of the main generation mechanisms of strong pathogenicity and drug resistance of pathogenic microorganisms. According to the difference between self-induced signal molecules and induction mechanisms, the QS system of bacteria can be divided into three types, namely, oligopeptide signal molecules (AIPs) of gram-positive bacteria, N-acyl-homoserine lactone signal molecules (AHLs) of gram-negative bacteria, and AI-2 signal factors common to interspecific cell communication.

Bacterial quorum sensing is a phenomenon that when a bacterial colony reaches a certain growth density, a cell body promotes specific gene expression and regulates and controls various life activities of the bacterial colony through secretion and action of self-induced molecules. The formation of staphylococcus epidermidis biofilm is closely related to the formation of Polysaccharide adhesin (PIA) between bacteria, and a strain with the PIA deleted can generate Adhesion obstacle and cannot form biofilm. Meanwhile, the quorum sensing of AIP as a self-induction signal molecule is a main quorum sensing system of staphylococcus epidermidis and gram-positive bacteria, and finally regulates and controls the secretion of PIA, the formation of a biofilm and the generation of virulence factors through the expression regulation and control of SarA on icaADBC.

AI-2 is a quorum sensing of signal molecules among bacteria, widely existing in gram-positive bacteria and gram-negative bacteria, and is a main way for information exchange among bacteria of different species. The current research shows that a common signal communication mode exists between the same strain and different strains, namely a molecular AI-2 mediated information communication mode between bacteria, and the channel is mainly to generate AI-2 molecules under the catalysis of LuxS protease encoded by LuxS gene, and regulate the secretion of polymers (EPS) of the biofilm to regulate the adhesion of the bacterial biofilm.

With the wide clinical application of broad-spectrum antibacterial drugs, the problem of drug resistance of opportunistic pathogenic bacteria is increasingly serious, so that the treatment capacity of common antibacterial drugs on skin soft tissue infection caused by bacteria is greatly reduced, and the desire of vast consumers on pursuing health, green and sustainability cannot be met. The flora quorum sensing inhibitor can well inhibit signal paths among flora, block the connection among flora and effectively inhibit the over-proliferation of bacteria to a certain extent. Meanwhile, because the traditional Chinese medicine components have higher safety and lower drug resistance compared with chemical medicines, the search for new antibacterial drugs or antibacterial drug compositions from the traditional Chinese medicine components has important development value and significance.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art. Therefore, the antibacterial composition provided by the invention can effectively inhibit the quorum sensing of the drug-resistant staphylococcus epidermidis, is safer and non-irritant to human bodies, has no side effects, and can be used for preparing skin external products related to skin soft tissue infection caused by the hyperproliferation of the drug-resistant staphylococcus epidermidis.

The invention also provides application of the antibacterial composition in preparation of a staphylococcus epidermidis inhibitor.

The invention also provides application of the antibacterial composition in preparing a medicament for treating and/or preventing related Skin Soft Tissue Infection (SSTI) diseases caused by excessive proliferation of staphylococcus epidermidis.

The invention also provides application of the antibacterial composition in preparation of an inhibitor for a quorum sensing system of a flora.

In one aspect of the present invention, there is provided an antibacterial composition comprising fraxinellone, dictamnine and thujone.

Further, the antibacterial composition comprises the following components in percentage by mass: 30-45% of fraxinellone, 25-35% of dictamnine and 25-35% of cypress ketone.

Preferably, the following components are contained in mass percent: 41% of fraxinellone, 27% of dictamnine and 32% of cypress ketone. In another aspect of the invention, there is provided the use of the antibacterial composition in the preparation of an inhibitor of staphylococcus epidermidis.

Further, the staphylococcus epidermidis capable of being inhibited by the staphylococcus epidermidis inhibitor is drug-resistant staphylococcus epidermidis. The antibacterial composition has good inhibitory effect on drug-resistant staphylococcus epidermidis.

Further, the staphylococcus epidermidis inhibitor can inhibit a quorum sensing system of staphylococcus epidermidis.

Further, the staphylococcus epidermidis inhibitor can inhibit the biofilm formation of staphylococcus epidermidis.

Wherein, the staphylococcus epidermidis can be staphylococcus epidermidis in different growth stages.

In another aspect of the present invention, there is provided a use of the antibacterial composition in the preparation of a medicament for treating and/or preventing related soft tissue infection (SSTI) diseases caused by excessive proliferation of staphylococcus epidermidis.

The term "treating", as used herein, includes alleviating, inhibiting or ameliorating the symptoms or conditions of a disease; inhibiting the generation of complications: ameliorating or preventing underlying metabolic syndrome; inhibiting the development of a disease or condition, such as controlling the development of a disease or condition; alleviating the disease or symptoms; regression of the disease or symptoms; alleviating a complication caused by the disease or symptom, or preventing or treating a symptom caused by the disease or symptom. As used herein, administration can result in an improvement in a disease, symptom, or condition, particularly an improvement in severity, delay in onset, slow progression, or decrease in duration of a condition. Whether fixed or temporary, sustained or intermittent, can be attributed to conditions associated with administration.

Further, the related skin soft tissue infection diseases comprise wound infection, epidermal abscess, cellulitis, chronic wound ulcer and the like.

Preferably, the drug-resistant staphylococcus epidermidis growth-related disease is wound infection.

Further, the dosage form of the medicine is an external preparation.

Preferably, the external preparation comprises a solution, a powder, a lotion, a tincture, a cream, an oil, a plaster, or the like.

In yet another aspect of the present invention, there is provided the use of the antibacterial composition for the preparation of an inhibitor of quorum sensing systems.

Further, the inhibitor of the quorum sensing system can inhibit a key gene sarA mediated by self-induced oligopeptides (AIP).

Further, the flora quorum sensing system inhibitor LuxS/AI-2 mediates the expression of a quorum sensing system key gene luxS.

In some embodiments of the invention, research results show that the antibacterial composition can significantly inhibit the expression of self-induced oligopeptide (AIP) -mediated quorum sensing system key gene sarA and LuxS/AI-2-mediated quorum sensing system key gene luxS, has statistical difference (P < 0.05), is nontoxic and non-irritant, and can be used for preparing a medicament-resistant staphylococcus epidermidis quorum sensing inhibitor and a skin external product for preventing or treating diseases related to skin soft tissue infection caused by hyperproliferation of medicament-resistant staphylococcus epidermidis.

Because the quorum sensing system is widely existed in microorganisms such as bacteria, fungi and the like, and regulates and controls various physiological functions, the quorum sensing system can effectively improve the stress resistance and environmental adaptability of microbial populations and is also one of the main generation mechanisms of strong pathogenicity and drug resistance of pathogenic microorganisms; therefore, the inhibition of the quorum sensing system of the bacterial colony can inhibit the physiological regulation of bacteria, including the inhibition of the production of exopolysaccharides, effector proteins, spores, the formation of biofilms and the like, can reduce the stress resistance and environmental adaptability of microbial populations, and can inhibit the strong pathogenicity and drug resistance of pathogenic microorganisms.

In still another aspect of the present invention, there is provided an epidermal staphylococcus inhibitor comprising the antibacterial composition.

In still another aspect of the present invention, there is provided a medicament for treating and/or preventing related skin soft tissue infection diseases caused by excessive proliferation of staphylococcus epidermidis, comprising the antibacterial composition.

In yet another aspect of the present invention, there is provided an inhibitor of quorum sensing systems, comprising the antibacterial composition.

The invention has the following beneficial effects:

the antibacterial composition has a remarkable inhibiting effect on staphylococcus epidermidis, shows that the antibacterial composition has a good inhibiting effect on staphylococcus epidermidis biofilms at different growth stages, has a remarkable inhibiting effect on a flora quorum sensing system of staphylococcus epidermidis, and can inhibit a quorum sensing system key gene sarA mediated by self-induced oligopeptides (AIP) and a quorum sensing system key gene luxS mediated by LuxS/AI-2. Meanwhile, the raw materials of the antibacterial composition are derived from natural traditional Chinese medicines, are safe, have no stimulation and side effects, have remarkable prevention and treatment effects on skin and soft tissue infection related diseases caused by hyperproliferation of the drug-resistant staphylococcus epidermidis, and can be applied to preparation of drug-resistant staphylococcus epidermidis quorum sensing inhibitors and drugs for preventing and treating skin and soft tissue infection related diseases caused by hyperproliferation of the staphylococcus epidermidis.

The antibacterial composition is a natural extract, has a good inhibiting effect on quorum sensing among drug-resistant staphylococcus epidermidis floras, has no toxic or side effect, is a safe and effective quorum sensing inhibitor with a Chinese herbal medicine source, has the effects of treating skin soft tissue infection diseases caused by staphylococcus epidermidis, does not have skin irritation and sensitization problems, and has important development value and significance.

The antibacterial composition has good antibacterial effect on staphylococcus epidermidis, particularly drug-resistant staphylococcus epidermidis, and the mechanism is that the expression of a quorum sensing system key gene sarA mediated by self-induced oligopeptide (AIP) and a quorum sensing system key gene luxS mediated by LuxS/AI-2 is inhibited, so that a quorum sensing system of pathogenic bacteria is inhibited, and the formation of a bacterial surface biofilm is inhibited. The antibacterial composition has the advantages of no toxicity, no irritation, etc. due to its component sources similar to those of natural Chinese medicinal plants.

In particular embodiments, the antimicrobial compositions of the present invention may be used alone or in combination with a number of other chemicals. Whether or not the chemical substance has biological activity or a function of treating a disease, the chemical substance includes one or more of a pharmaceutically acceptable carrier, a food, a natural product, a chemically synthesized drug, or a human drug.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Drawings

FIG. 1 is a graph showing the effect of the antimicrobial composition of example 3 on the adhesion rate of Staphylococcus epidermidis SEP06 biofilm (3h, initial adhesion period);

FIG. 2 is a graph showing the effect of the antimicrobial composition of example 3 on the adhesion rate of Staphylococcus epidermidis SEP06 biofilm (8h, logarithmic growth phase);

FIG. 3 is a graph showing the effect of the antimicrobial composition of example 3 on the adhesion rate of Staphylococcus epidermidis SEP06 biofilm (24h, mature stage);

FIG. 4 is a graphic representation of experimental cell images showing the effect of the antimicrobial composition of example 3 on the removal of the biofilm of Staphylococcus epidermidis SEP06(3h, 8h, 24 h); (e) graph of cell imaging of different concentrations of antibacterial composition in initial adhesion stage; (a) graph of cellular imaging of different concentrations of antimicrobial compositions at logarithmic growth phase; (i) graph of cell imaging of different concentrations of antibacterial composition at maturation stage;

FIG. 5 is a graphical representation of the vasoreactivity effect on chick embryo chorioallantoic membrane (CAM) in example 5 of the present invention; wherein, (A) 0.9% NaCl solution group, (B) 1% Texpon ASV solution group, (C) antibacterial composition group, and the vascular response effect of each group of CAMs after being exposed to CAM for 5min are shown.

Detailed Description

In order to explain technical contents, achieved objects, and effects of the present invention in detail, the following description is made with reference to the accompanying drawings in combination with the embodiments.

Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and materials used in the following examples are commercially available.