KR20110125118A - Antifungal compound and use thereof - Google Patents

Abstract

PURPOSE: An antifungal composition containing a compound is provided to ensure antifungal and antibacterial activities with safety to human body. CONSTITUTION: A compound with antifungal and antibacterial activities is denoted by chemical formula 1. An antifungal composition contains the compound as an active ingredient. The composition has an antifungal activity against Aspergillus sp., Cladosporium sp., Penicillium sp., or Candida sp. A preservative contains the compound as an active ingredient. A composition for treating or preventing fungal diseases contains the compound as an active ingredient. The compound is delta-dodecalactone and is isolated from supernatant of Lactobacillus culture liquid.

Description

Antifungal Compounds and Their Uses {ANTIFUNGAL COMPOUND AND USE THEREOF}

The present invention relates to compounds having antifungal activity and the use of such compounds.

It is reported that between 5% and 10% of world food production is lost to fungal decay. In particular, Aspergillus or Fungi, such as Penicillium , are known to be a major cause of decay or rot of various types of food or feed that require long-term storage. In addition, the fungal toxins produced by these fungi are known to cause serious diseases in animals, especially humans, who have consumed rot or decayed foods.

Physical and chemical methods have been proposed as methods for inhibiting the growth of these fungi for long term storage of food or feed. The physical method is a method of treating a storage object by a process such as freezing, drying, heat treatment or low temperature storage, and the chemical method is a method of adding a synthetic additive or synthetic preservative such as a preservative to the storage object. However, in recent years, consumer's objection to artificial synthetic preservatives or chemical synthetic additives is increasing, and along with the excellence of quality, consumers' demands for foods containing no preservatives or minimal processing such as organic foods are increasing. . In response to these consumer demands, there is a growing interest in natural preservatives as an alternative to chemical preservatives.

In addition, the Green Round Convention on Environmental Protection and Ecosystem Conservation has become an international concern, and overuse of pesticides is becoming a serious problem. Therefore, in order to overcome the problem of environmental pollution for the existing pesticides or preservatives, the demand for the development of new fungicides that can solve the problems of the existing antifungal agent is increasing. In connection with the development of these new fungicides, there is an increasing need for the development of pollution-free biopides that replace existing chemical preservatives, chemical pesticides or artificial synthetic fungicides.

Lactic acid bacteria (LAB) have been reported to produce a variety of antimicrobial substances such as lactic acid, acetic acid, hydrogen peroxide, diacetyl, carbon dioxide, and bacteriocin, but most of these findings are focused on antibacterial effects. However, very few studies have been conducted on lactic acid bacteria having antifungal performance, and studies on the antifungal substance itself and its mechanisms derived from lactic acid bacteria are still insignificant.

Kimchi, a traditional Korean food, is fermented by various microorganisms after mixing cabbage and radish with spices, spices, and salted fish, and is known as a food using lactic acid bacteria fermentation. Kimchi, a fermented food, contains a variety of microorganisms, and it is speculated that among these microorganisms, various substances acting as natural preservatives are produced due to lactic acid bacteria. However, these various substances are often not precisely identified as specific substances, and most of them study whether there is sufficient antibacterial or antifungal effect as well as a mechanism that acts as a natural preservative and commercially available. It is not.

An object of the present invention is to provide a compound having antibacterial and antifungal activity isolated from lactic acid bacteria which are GRAS (Generally Recognized As Safe) microorganisms isolated from kimchi.

Another object of the present invention is the use of the compound having antifungal activity, specifically, the use of a compound based on the antifungal activity and antibacterial activity and a product applied thereto, for example, a preservative comprising a compound having the antifungal activity, a functional food , Cosmetic compositions, to provide a composition for the treatment or prevention of fungal dermatitis comprising the substance as an active ingredient.

In order to achieve the above object, the present invention provides a compound having antibacterial and antifungal activity. More specifically, the compound of the present invention may have the following formula (1) having antibacterial and antifungal activity.

[Formula 1]

In addition, the present invention provides a composition having an antimicrobial activity and an antifungal activity comprising the compound represented by Formula 1 as an active ingredient.

The present invention also provides a preservative comprising the composition as an active ingredient. The preservative may be applied as a preservative used for the storage of stored foods including agricultural products, food additives or food preservatives used in foods, cosmetic additives used in cosmetics, pharmaceutical additives used in cosmetics or pharmaceuticals, or preservatives for pharmaceuticals. Can be.

The present invention also provides a cosmetic composition comprising the composition as an active ingredient, more specifically a cosmetic composition having antifungal efficacy.

In addition, the present invention provides a composition for the treatment or prevention of fungal diseases comprising the composition as an active ingredient.

Hereinafter, the present invention will be described in more detail.

The inventors of the present invention, while researching to find antifungal substances having antifungal activity from lactic acid bacteria, which are generally recognized as safe (GRAS) microorganisms, which are not a safety issue, unlike other microorganisms, which have been mainly reported in the past, the inventors of the present invention Lactobacillus platarum AF1 isolated from kimchi plantarum AF1, KFCC-11340) was isolated from the culture supernatant of the antifungal activity, and the isolated material was confirmed to have an antifungal activity against a variety of fungi, including Candida fungi belonging to fungi and yeast, the antifungal activity The present invention was completed by confirming the spectrum and antibacterial spectrum.

In the present invention, "antibacterial" means an action to limit, reduce or eliminate bacteria, harmful microorganisms or pathogenic microorganisms present in the environment, and may be used in the same sense as the antibacterial. "Bacteria" is a concept that includes both eubacteria and archeobacteria.

In the present invention, "antifungal" means an action intended to limit, reduce or eliminate fungi present in the environment. "Fungus" means all fungi belonging to the fungus, including Aspergillis and Candida.

The environment is a concept encompassing all cases in which the bacteria and / or fungi can grow, and a plant or a part of a plant including the grains in which the bacteria and / or fungi can grow, animals including humans, food, medicines and cosmetics It includes. The environment can be a solid, liquid or gas.

In the present invention, the "antimicrobial composition" may mean the same as an antibiotic, which is a generic term for an antimicrobial agent, and may mean the same as an antimicrobial agent, a fungicide, a preservative, a preservative or an antifungal agent, and preferably the development of fungi and bacteria. Means a substance that can inhibit or inhibit life and life functions.

In the present invention, the "preservative" means the deterioration, decay, discoloration and chemical of preservative substances such as food, feed, cosmetics and pharmaceuticals, such as to prevent the preservation substances from being rotted by bacteria or microorganisms or exposed to air. It is used to prevent change and to maintain the functionality and freshness of the material to be preserved. In a narrow sense, it is used to enable long-term storage of foods, feeds, food ingredients, processed foods, cosmetics or medicines, including grains, mainly through the inhibition or sterilization of harmful microorganisms or decaying microorganisms such as bacteria, fungi or yeasts. It means to be.

In the present invention, "food" means a generic term for all ingredients that humans can consume or humans can eat. The food is a concept that includes all of the raw materials of natural or processed products and processed products containing one or more nutrients and no harmful substances, for example, processed food, fish meat products, tofu, jelly, health supplement food And seasonings, bakery and confectionery, dairy products, pickles, fermented foods or beverages.

In the present invention, "feed" means all substances provided to supply organic or inorganic nutrients necessary for maintaining livestock life. The feed includes livestock feed, and its use or formulation is not limited. For example, it includes a feed, rich feed, supplemental feed, protein feed, starch feed, fat feed, fiber feed or mineral feed. In addition, the formulation of the feed may be a solid (solid) or liquid (liquid), but is not limited thereto.

The present invention relates to compounds with antifungal activity, more particularly compounds with antibacterial and antifungal activity.

More specifically, the compound is a compound represented by the following formula (1).

[Formula 1]

The compound represented by Chemical Formula 1 is Lactobacillus Platarum AF1 ( Lactobacillus) isolated from kimchi plantarum AF1, KFCC-11340) may be isolated from the culture supernatant. The Lactobacillus platarum AF1 was deposited with the Korea Microorganism Conservation Center on November 6, 2004 and received accession number KFCC-11340. The Lactobacillus platarum AF1 was deposited with the Korea Microorganism Conservation Center on November 6, 2004 and received accession number KFCC-11340. The Lactobacillus plantarum AF1 can be grown in a conventional lactic acid bacteria medium. When the Lactobacillus plantarum AF1 is incubated at 30 ° C using a liquid medium, it may reach a growth stop after 16 hours to 20 hours of incubation. In order to obtain the culture supernatant, the Lactobacillus plantarum AF1 may be cultured under anaerobic conditions at a culture temperature of 25 ℃ to 35 ℃.

The method for separating the compound having an antifungal activity represented by Formula 1 from the culture supernatant of the Lactobacillus platarum AF1 may use a method for separating an antifungal activity substance from a conventional microbial culture supernatant. The substance having antifungal activity isolated by the above method was confirmed the structure of the compound using GC-MS. As a result of confirming the structure, the structure of the compound represented by Chemical Formula 1 was confirmed, and it was confirmed that the compound represented by Chemical Formula 1 was delta-dodecaractone through the structure of the compound represented by Chemical Formula 1.

The present inventors separated the material having antifungal activity from the culture of lactic acid bacteria, more specifically, lactic acid bacteria through the embodiment of the present invention, it was confirmed for the first time that the delta-dode caractone has antifungal activity. The compound represented by the formula (Delta) -docatelactone was confirmed to have antifungal activity against various fungi as well as antibacterial activity against bacteria that are various pathogenic microorganisms.

In addition, the compound represented by Formula 1 includes culturing the Lactobacillus platarum AF1 to obtain a culture solution, obtaining a culture supernatant from the culture medium and separating the compound represented by the formula 1 from the culture supernatant solution It can be prepared by the method.

In the obtaining of the culture solution, the Lactobacillus platarum AF1 may be cultured in a liquid medium, preferably, lactic acid bacteria medium, the culture conditions may be anaerobic conditions, and the culture temperature conditions may be 25 ° C to 35 ° C. . In addition, the step of obtaining the culture supernatant may be performed by a method of obtaining the supernatant by centrifuging the culture solution using a centrifuge, and may further perform the process of filtration or sterilization using a filter paper or a filter. In addition, the step of separating the compound may be performed by a separation purification process, for example, solid phase extraction (SPE) and / or HPLC for the culture supernatant.

In addition, the present invention relates to the use of a compound represented by the formula (1) having the antibacterial and antifungal activity. The use may be a use using antibacterial and antifungal activity.

Specifically, the use is a preservative used for the storage of stored foods, including agricultural products, specifically food additives or food preservatives used in food, cosmetic additives used in cosmetics or pharmaceutical additives or pharmaceuticals used in cosmetic preservatives or medicines It may be a preservative, a cosmetic composition, specifically a cosmetic composition having an antifungal effect, or a composition for treating or preventing a disease caused by a fungal disease or a fungal infection, including, but not limited to, the use of a preservative for the present invention. It may include any use that utilizes or applies the antifungal and antimicrobial effects of the compound.

The compound of Formula 1 was obtained by separating from the culture supernatant of the Lactobacillus platarum AF1 having antifungal activity, the compound of Formula 1 is a fungus that contaminates peanuts, corn and cereals, and has been reported to be a potent liver cancer-causing substance (Aflatoxin). aflatoxin) to create Aspergillus Playa bus (Aspergillus flavus), and this causes deterioration of stored grain fungus Aspergillus pneumonia pathogens in Phu called Aspergillus fumigatus seujeung to (Aspergillus fumigatus), Ochratoxin A (ochratoxin A) Aspergillus Petra key (Aspergillus capable of producing toxins that petrakii), dermal toxin (dermatoxin) and one kind of fungal poisoning can create a cholesteric Mato cystine (sterigmatocystin) that may cause, and a variety of food stores and cause Aspergillus nidyu Lance (Aspergillus that the corruption of the stored fodder nidulans ), the causative agent of dermatitis, Cladosporium gossypiicola), Penny rooms that corruption is a major cause of food cold storage Leeum rokwe FORT Tea (Penicillium roqueforti) and athlete's foot and yeast, including Candida albicans that causes various skin diseases (Candida albicans ) have been shown to have antifungal activity against.

In addition, the present invention relates to an antifungal composition comprising a compound represented by the following formula (1) as an active ingredient. The antifungal composition may have antifungal activity as well as antibacterial activity, specifically, antimicrobial activity against pathogenic microorganisms.

[Formula 1]

The antifungal composition may include 0.001 to 99.99% by weight, preferably 0.1 to 99% by weight of the compound represented by Formula 1, based on the total weight of the composition, and is effective according to the method of use and purpose of use of the antifungal composition. The content of the component can be properly adjusted.

The composition is a fungus, specifically Aspergillus sp., Cladosporium sp., Penicillium sp. And Candida sp. At least one fungus selected from the group consisting of: preferably Aspergillus flavus ), Aspergillus pumigatus fumigatus ), Aspergillus petraki petrakii), Aspergillus okra three mouse (Aspergillus ochraceus), Aspergillus nidyu lance (Aspergillus nidulans ), Cladosporium gossypiicola , Penicillium loqueforti roqueforti ) and Candida albicans may have antifungal activity against one or more fungi selected from the crowd.

Aspergillus plabus is a fungus found in almost all the world's soils. It contaminates cereals, including peanuts and corn, has strong toxicity, and is called aflatoxin, which acts as a carcinogen. Produces toxic substances. Thus, a substance having antifungal activity against the Aspergillus plabus can prevent contamination of cereals by the fungi and can treat and prevent diseases caused by the fungi.

Aspergillus pumigatus is a fungus found in soil, water, or various organic substances, and grows in lungs in which tissues are destroyed due to pulmonary tuberculosis, bronchiectasis, lung abscess, and the like. Aspergillosis induces fungal pneumonia. It can also be a fungal cause of storage grain decay. Accordingly, the substance having antifungal activity against Aspergillus fumigatus can prevent contamination of the stored grains by the fungus, and can treat and prevent diseases such as fungal pneumonia caused by the fungus.

Aspergillus petraki is a fungus capable of producing a toxic substance called okratoxin A, which causes decay or decay of organic matter. Thus, a substance having antifungal activity against Aspergillus petraki can treat and prevent diseases caused by the fungi and the okratoxin A produced by the fungi.

Aspergillus okrasus is a contaminant contaminating stored foods such as stored dried foods, dried fish, dried beans, beef jerky, dried fruits or nuts such as peanuts, hazelnuts, walnuts, kidney toxicity, immunosuppression, carcinogenesis, etc. It is a fungus that can produce ochratoxin, which has been shown to be a cause. Thus, a substance having antifungal activity against Aspergillus okraceus can prevent contamination of the stored food, in particular dry food, by the fungus, and can be induced by the okratoxin and the disease caused by the fungus. Can be treated and prevented.

Aspergillus nidurans are contaminants that contaminate stored cereals and animal feed, and are fungi that can produce mycotoxin. Thus, a substance having antifungal activity against Aspergillus nidurans can prevent contamination of the stored foods, in particular animal feed, by the fungi, and can be induced by the fungal poisons including allergic diseases, and The disease caused by the fungus can be treated and prevented.

The cladosporium gopicola has been reported as a causative agent of skin diseases including allergic, fungal infections, pneumonia and dermatitis. Therefore, the substance having antifungal activity against the Cladosporium gopicola can treat and prevent allergies, pneumonia, and skin diseases caused by the fungi.

The penicillium roquefort is reported to contaminate cold stored foods stored at low temperatures. Therefore, the substance having antifungal activity against the penicillium roquefort can prevent the contamination of the stored foods by the fungus, especially cold storage is required or mainly stored at low temperatures.

Candida albicans is a fungus that causes candidiasis or monolithosis, and refers to an incomplete fungus belonging to the genus Candida. When the oral mucosa is contaminated by the Candida albicans, oral candidiasis called thrush develops. In addition, when the candida albicans is inflamed by breeding in the vagina or vulva, candida vaginitis with the highest incidence of vaginitis occurs. Candida albicans is also a causative agent of superficial fungal disease or athlete's foot, which is a disease caused by infection of the fungus with the stratum corneum or nail. Accordingly, the substance having antifungal activity against Candida albicans can treat and prevent skin diseases caused by the fungus, specifically, superficial fungi, athlete's foot, oral candidiasis, and candida vaginitis.

In addition, the composition may further have antimicrobial activity against pathogenic microorganisms. More specifically, the composition is a bacterium, preferably Gram-positive bacteria and / or Gram-negative bacteria, more preferably Bacillus sp., Streptococcus sp., Staphylococcus sp., Enterococcus sp., Listeria sp. And Escherichia coli ) one or more bacteria selected from the group consisting of, more preferably Bacillus subtilis ( Bacillus subtilis ), Streptococcus mutans ), Staphylococcus aureus), Enterococcus faecalis (Enterococcus faecalis), L. monocytogenes to Ness (Listeria monocytogenes) and Escherichia coli (E. coli) as may have antibacterial activity against one or more bacteria selected from the consisting of a crowd.

In addition, the Streptococcus mutans as a causative agent of caries can be used for the purpose of treatment and prevention of caries, periodontitis and gingivitis, etc., the staphylococcus aureus is a causative agent of skin infection and pneumonia, As a result of pneumonia, pneumonia, empyema, respiratory infections, sepsis, bacteremia, infective endocarditis, enteritis, intestinal infections, intractable pressure sores or wound infections, substances having antibacterial activity against the bacterium are refractory pressure sores and impetigo It can be used for the treatment and prevention of skin diseases such as cellulitis, sepsis, pneumonia, etc. The Listeria monocytogenes and E. coli are representative food poisoning and food rot causing bacteria, and substances having antimicrobial activity against the bacteria are Can be used for the treatment and prevention purposes, to prevent the corruption of food or to preserve food To be used in the form of additives and the like.

While the compound of Formula 1 shows excellent antifungal activity, it is a component extracted from the culture supernatant of Lactobacillus platarum AF1, which is a lactic acid bacterium extracted from kimchi, and has an advantage of being safe for human body.

The antifungal composition or composition having antifungal activity and antimicrobial activity of the present invention can be directly applied to animals including humans to exhibit antimicrobial activity, and can be used in feed for feed animals to lower mortality and increase productivity of breeding animals. It can be used for the purpose of preventing secondary infection in humans that consume livestock animals. The animal is a biological group corresponding to plants, and mainly eats organic matter as nutrients, and means that digestion, excretion, and respiratory organs are differentiated, and may be preferably a mammal or a bird.

The antifungal composition or the composition having the antifungal activity and the antimicrobial activity may further include sugars, proteins, lipids, vitamins and minerals in addition to the compound represented by Chemical Formula 1.

The sugar may be appropriately selected according to the purpose and use thereof, and may be, for example, honey, dextrin, sucrose, palatinose, glucose, fructose, syrup, sugar alcohol, sorbitol, xylitol, maltitol, It is not particularly limited thereto. The protein may be appropriately selected depending on the purpose and use thereof. For example, animal-derived enzymes such as milk derived proteins such as casein and whey protein, soy protein, trypsin of these proteins, and pepsin. And hydrolysates by neutrases and alkalases, but are not particularly limited thereto. The lipid may be appropriately selected according to the purpose and use thereof. For example, the monohydric saturated fatty acid, sunflower oil containing polyunsaturated fatty acid, rapeseed oil, olive oil, safflower oil, corn oil Oil, soybean oil, palm oil (palm oil), palm oil and other plant-derived fats, such as middle-chain fatty acids, EPA, DHA, soybean-derived phospholipids, milk-derived phospholipids, but is not particularly limited thereto. The vitamins are not particularly limited, and the minerals may be appropriately selected depending on the purpose and use thereof. For example, potassium phosphate, potassium carbonate, potassium chloride, sodium chloride, calcium lactate, calcium gluconate, calcium pantothenate, and casein calcium , Magnesium chloride, ferrous sulfate, sodium hydrogen carbonate, but is not particularly limited thereto.

For any of the above saccharides, proteins, lipids, vitamins and minerals, as long as the antimicrobial properties of the composition are maintained, not limited to the above embodiments and may include other ingredients, each content is not limited as long as the antimicrobial properties are maintained Preferably, it may be 0.1 to 15% by weight, preferably 0.5 to 10% by weight relative to the total composition.

The antifungal composition or the composition having antifungal activity and antimicrobial activity may be used for various purposes and uses requiring antifungal activity or antifungal activity and antimicrobial activity, and specifically, prevention of diseases caused by infection of pathogenic microorganisms, in particular pathogenic fungi. Or an active ingredient of a cosmetic composition including a therapeutic composition or a cleaning composition for the purpose of alleviating a disease caused by an infection of the pathogenic fungus, a food preservative, a cosmetic preservative, a feed preservative or a medicine for preventing corruption by the pathogenic microorganism. The use of a preservative including a preservative, the use of additives for animal feed for preventing the decay of the feed by the pathogenic microorganisms, or may be used as an active ingredient of pesticides, including agricultural fungicides.

In addition, the present invention provides a composition for treating or preventing fungal diseases comprising, for example, a compound represented by Formula 1, the antifungal composition, or a composition having the antifungal activity and antibacterial activity as an active ingredient.

The fungal disease means a disease caused by infection of pathogenic fungi. The pathogenic fungi may be at least one selected from the group consisting of Aspergillus fungi, Cladosporium fungi, Penicillium fungi and Candida fungi, for example Candida albicans.

The fungal disease is a disease caused by an infection of the fungus, which is caused by a fungal poison such as Mycotoxin, specifically, aflatoxin or okratoxin, secreted by the fungus itself or fungi. It may be a disease, for example, at least one selected from the group consisting of fungal dermatitis, athlete's foot and fungal pneumonia.

The composition for the treatment or prevention of fungal diseases of the present invention is a carrier or excipient pharmacologically usable according to the formulation and method of use in addition to the compound represented by the formula (1), the antifungal composition or a composition having the antifungal activity and antibacterial activity. In addition, the composition of the present invention may be administered alone or in combination with other pharmaceutically active ingredients for enhancing efficacy as an antifungal activity or as a therapeutic agent for diseases caused by infection of fungi. In this case, the content of the active ingredient in the composition for treating or treating the fungal disease may be 0.001 to 99% by weight. If the content of the active ingredient in the composition is less than 0.001% by weight may require a large amount of administration for effective efficacy, if the amount exceeds 99% by weight may be constant compared to the amount may be uneconomical. In addition, the composition for the treatment or prevention of fungal diseases according to the method of use and purpose of use can appropriately adjust the content of the active ingredient.

The composition for treating or preventing fungal diseases of the present invention may be administered orally or parenterally, and its formulation may vary depending on the method of use, and is not limited thereto.

Examples of the formulation of the composition for the treatment or prevention of fungal diseases include PLASTERS, GRANULES, LOTIONS, POWDERS, Syrups, LIQUIDS AND SOLUTIONS, Aerosols AEROSOLS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS, SUSPENSIONS, INFUSIONS, TABLETS, INJECTIONS, CAPSULES, AND PILLES PILLS). Further compositions for the treatment or prevention of fungal diseases can be prepared using methods known in the art or as disclosed in Remington's Pharmaceutical Science (Recent Edition, Mack Publishing Company, Easton PA). And preferably formulated.

Excipients such as pharmacologically acceptable carriers or diluents, preservatives, stabilizers, wetting agents, emulsifiers, solubilizers, sweeteners, colorants, osmotic pressure regulators, antioxidants, etc. may use conventional materials according to the formulation, and when formulated, fillers , Extenders, wetting agents, disintegrating agents or surfactants may be used. Representative diluents or excipients include water, dextrin, calcium carbonate, lactose, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyloleate, liquid paratin and saline.

Specifically, in the case of oral administration depending on the formulation, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. which are commonly used may be used. Suspensions, emulsions, lyophilized preparations, suppositories, and the like can be used. The non-aqueous solvent or suspending agent may be, for example, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like, and suppositories may include, for example, Utopsol, macrogol, tween 61 , Cacao butter, laurin butter or glycerogelatin and the like.

In addition, the composition for the treatment or prevention of fungal diseases of the present invention may be used in admixture with various carriers acceptable as medicaments, such as physiological saline or organic solvents, and to increase the stability or absorption of glucose, sucrose or dex It can be used with carbohydrates such as tran, ascorbic acid or antioxidants such as glutathione, chelating agents, small molecule proteins or other stabilizers.

Subjects of the composition for the treatment or prevention of fungal diseases of the present invention may be animals, including echinoderm, crustaceans, molluscs, fish, amphibians, reptiles, birds and mammals, preferably birds and / or mammals, more Preferably, it may be a livestock capable of breeding humans, pigs, cattle, goats, and the like and / or poultry such as pheasants and chickens.

The dosage or amount of the composition for the prophylaxis or treatment of the composition for the treatment or prophylaxis of fungal diseases of the present invention may be determined by weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease. It is desirable to determine and take into account, for example, the effective daily dosage is typically 10 to 5000 mg / kg or 100 to 3000 mg / kg for adults (70 kg) and can be administered 1 to several times a day, preferably Preferably 1-3 times daily.

Since the dosage may vary depending on various conditions, it will be apparent to those skilled in the art that the dosage may be added or subtracted, and thus the dosage does not limit the scope of the present invention in any aspect. The frequency of administration can be administered once a day or divided into several times within the desired range, the administration period is not particularly limited.

The composition for the treatment or prophylaxis of fungal diseases of the present invention not only provides an excellent antifungal effect and an antimicrobial effect of preventing or treating the infection, but also is isolated from a substance that generates GRAS microorganisms, lactic acid bacteria, so the safety is positive, You can take it.

In addition, the present invention is an example for the prevention of at least one selected from the group consisting of fungal dermatitis and athlete's foot comprising a compound represented by the formula (1), the antifungal composition or a composition having the antifungal activity and antibacterial activity as an active ingredient Or it provides a cleaning composition for improvement. The detergent composition may be applied to the skin or oral cavity or to the nail or toenail.

In addition, the present invention prevents or improves at least one selected from the group consisting of fungal dermatitis and athlete's foot, including the compound represented by Formula 1, the antifungal composition or a composition having the antifungal activity and antibacterial activity as an active ingredient. It provides a cosmetic composition for. The cosmetic composition may be applied to the skin, nails or toenails.

The cleaning composition comprising the compound represented by the formula (1), the antifungal composition or the composition having the antifungal activity and the antimicrobial activity as an active ingredient is a material commonly used in the preparation of the active ingredient, for example, a surfactant, tax information It may further include moisturizers such as premature agents, pH adjusters, natural moisturizing ingredients and natural vegetable oils, antioxidants, metal ion sequestrants, preservatives, solvents, flavoring agents and pigments. In addition, the antifungal effect and antimicrobial effect of the present invention can be maintained, and may further include additional antifungal substances or antimicrobial substances within a range that does not irritate the human body.

The detergent composition is a composition used to remove oily and aqueous pollution or wastes attached to the skin surface with moderate foaming power in cold water and hot water. Recently, moisture is evaporated while supplying moisture to the skin. Additional functions such as imparting a moisturizing effect, increasing skin flexibility, or increasing sebum removal ability may be further provided.

In general, the infection of the skin is caused by the penetration of bacteria into the sebum in the pores, or caused by allergic factors, when the cleaning composition using the cleaning composition of the present invention by obtaining the antifungal effect and antibacterial effect according to the active ingredient skin The condition can be improved. In addition, the role of removing the waste or sebum in the cleaning composition is mainly a surfactant, and the prevention of infection by fungi and bacteria is carried out by the active ingredient, since the conventional fungicides or antimicrobial materials are mainly chemical synthetic materials For example, when used on skin sensitive to chemical synthetic materials or used excessively over a certain level, the skin may be damaged by contact with the skin, and there is a problem in that it cannot effectively prevent infection by fungi.

In the cleaning composition of the present invention, the compound represented by Chemical Formula 1, which prevents infection by fungi and bacteria, is excellent in antifungal effect and is recognized as safe as it is isolated from a substance that generates GRAS microorganism, lactic acid bacteria. It has the advantage of stimulation.

The surfactant may be any one selected from the group consisting of anionic surfactants, nonionic surfactants, cationic surfactants and amphoteric surfactants.

The anionic surfactant is, for example, ammonium lauryl sulfosuccinate, ammonium lauryl sulfate, sodium cocoyl isionate, sodium lauryl isceionate, sodium lauryl sulfate, triethanolamine lauryl sulfate or sodium lauryl ether sulfate And the like. The nonionic surfactant may be, for example, polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyoxyethylene, polysorbate 80, hardened castor oil derivative, fatty acid diethanolamide, glyceryl stearate, or the like. The cationic surfactant may be, for example, a tertiary aliphatic amine salt, alkyltrimethylammonium chloride or dialkaldimethylammonium chloride. The amphoteric surfactant may be, for example, betaine, amidebetaine type, sulfobetaine type, or steartrimonium chloride.

For example, the pH adjusting agent may be organic acid, such as citric acid, succinic acid, tartaric acid, lactic acid, malic acid, fumaric acid, maleic acid, glycolic acid, hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, or the like, or a sodium salt, potassium salt, or ammonium salt thereof. Triethanolamine, sodium chloride, sodium hydroxide or potassium hydroxide, and the like.

The moisturizing agent is a conventional oil used for moisturizing, for example, vegetable oil or liquid paraffin, ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, sorbitol, Collagen and the like. The moisturizing agent may be contained in an amount of 0.1 to 10 parts by weight based on the total weight of the composition, but is not limited thereto.

The preservative may be, for example, benzoic acid, paraoxybenzoic acid ester, a mixture of methylchloroisothiazolinone, phenoxyethanol, DMDM hydantoin, methyl paraben, ethyl paraben, or propyl paraben.

The solvent may be, for example, purified water, ethanol, Tween 20, cyclomethicone, mineral oil, or dimethicone, and the anti-corrosive agent and the pigment may be selected by a person having ordinary skill in the art to which the present invention belongs. The components used in the composition can be used.

The detergent composition may also be formulated as a liquid, solid soap or powder.

The cosmetic composition may be for application to the skin, nails and / or toenails.

The cosmetic composition may be prepared in the form of a cream, lotion, gel, pack, liquid, spray, water dispersion, powder or oil. The composition of each formulation may contain a variety of bases and additives necessary for the formulation of the formulation, and the type and amount of these components can be easily selected by those skilled in the art within the range that can maintain the effect of the active ingredient. have.

When the formulation of the cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silita, talc or zinc oxide, etc. are further included. can do.

When the formulation of the cosmetic composition is a powder or a spray, it may further include lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder, in particular when the formulation is a spray, additionally chlorofluoride Propellants such as rockcarbon, propane, butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or emulsion, it may include a solvent, a solubilizer or an emulsifier, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan and the like.

When the formulation of the cosmetic composition is a suspension, a liquid diluent such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like may further be included.

In addition, the present invention provides a composition for animal feed comprising the compound represented by the formula (1), the antifungal composition or a composition having the antifungal activity and antibacterial activity as an active ingredient.

The animal feed composition may further include grain, vegetable protein feed, animal protein feed, sugar or dairy product in the active ingredient. The grains may be specifically milled or crushed wheat, oats, barley, corn and rice, and the vegetable protein feed may be specifically based on rapeseed, soybean and sunflower, the animal protein feed specifically May be blood meal, meat meal, bone meal and fish meal, and the sugar or dairy product may specifically be a dry ingredient consisting of various powdered milk and whey powder.

The animal feed composition may further be used with ingredients such as nutritional supplements, digestion and absorption enhancers, growth promoters or disease prevention agents.

The active ingredient included in the composition for animal feed of the present invention may vary depending on the purpose and conditions of use of the feed. For example, the active ingredient may be included in an amount of 0.001 g to 100 g based on 1 kg of the final produced feed.

In addition, the feed composition may be made of a viscous granulated or granular material depending on the degree of grinding of the components, the composition may be supplied in a mesh or formed into a desired separated shape for further processing and packaging. It may be subjected to a pelletizing, expanding or extruding process for storage, and preferably excess water may be dried off for ease of storage.

The present invention also provides a cleaning agent comprising the compound represented by the formula (1), the antifungal composition or a composition having the antifungal activity and antibacterial activity as an active ingredient. The cleaner may be a detergent for clothing, a dish cleaner, a food cleaner, or a household appliance cleaner, but is not limited thereto. Compound represented by the formula (1) is not only excellent in the antifungal effect and antibacterial effect, but also safety is recognized as isolated from the material that produces the GRAS microorganism lactic acid bacteria, washing or cleaning various household products, including kitchenware or food washing It can be used for the purpose of imparting antifungal and antibacterial properties.

The cleaning agent may further include an appropriately selected additive according to the purpose of use and use thereof, and may be mixed with other active ingredients such as a commonly used cleaning agent, and additives such as colorants, surfactants, and preservatives. The detergent may be prepared by powdering, granulating, tableting or liquefying according to the purpose and use thereof, and a conventional method may be used for packaging for commercialization.

In addition, the amount of the active ingredient used in the cleaning agent may be suitably used according to the purpose of use, application form, application site, target object, and the like, and may include, for example, 0.001 to 99 parts by weight based on the total composition of the antimicrobial composition. The content is not limited thereto.

In addition, the present invention provides a preservative including, as an active ingredient, a compound represented by Formula 1, the antifungal composition, or a composition having the antifungal activity and the antimicrobial activity. The preservative may be used for the purpose of preserving food, stored food including grains, cosmetics and / or pharmaceuticals. The compound represented by Formula 1 is excellent in antifungal effect and antibacterial effect, and safety is recognized as being isolated from the material which produces GRAS microorganism lactic acid bacteria, there is an advantage that can be added to various preservation materials.

The preservative may be applied as food additives, cosmetic additives or pharmaceutical additives.

The preservative may be used in a range in which the active ingredient does not change the effect intended by the preservative substance such as food, cosmetics or medicines and does not affect the quality of the preservative substance. It may be applied in an amount of 0.001 to 10 parts by weight as a standard, but is not limited thereto.

The preservative may further include an antioxidant such as amygalate, tocopherol, butylhydroquinone, etc., in addition to the active ingredient, depending on the purpose of use of the preservative, and the additionally included components include the purpose of the active ingredient, that is, antifungal and antibacterial properties. It may be included in an amount within a range that does not impair the purpose.

For example, when the preservative is used in food, it is also called a food preservative, and in the case of the food preservative, in addition to the active ingredient, a preservative or a fungicide that suppresses the growth of mold may be used together. . As the preservative, for example, sorbic acid, dihydroacetic acid, etc. may be used, and as the preservative, parahydroxybenzoic acid esters, propionate, and the like may be used.

In addition, the preservative may be used in food storage processes such as cereals. For example, since a certain amount of grain is sprayed to prevent corruption and contamination of the grain by fungi, food shortage problems recognized as urgent problems around the world and problems associated with food transportation to food shortage areas It is expected to be solved. When applied for the above purpose, the present invention can be used as an antiseptic or preservative containing the active ingredient.

The compound of the present invention is a fungus that contaminates peanuts, corn and cereals, causes Aspergillus plabus to produce aflatoxin, a potent liver cancer-causing substance, and causes decay of stored grains, and causes fungal pneumonia called aspergillosis. Lactobacillus platarum, which has excellent antifungal activity against phosphorus Aspergillus pumigatus, Candida albicans, a causative agent of various skin diseases, has antimicrobial activity against pathogenic microorganisms, and is isolated from kimchi belonging to the GRAS microorganism lactic acid bacteria. As a component extracted from the culture supernatant of AF1, it is also very safe for the human body, and various applications requiring antifungal activity and / or antimicrobial activity as well as antibiotics are one example, medicaments for the treatment of diseases caused by fungal infection, prevention of the fungal infection and Cosmetics, cleaning compositions, and preservation for improvement First, it can be used for various uses, such as food additives for cosmetic purposes, cosmetic preservatives, medical additives, and the like, and can prevent fungal contamination of cereals, so the industrial effect will be very large.

1 is the result of the first injection of prep-HPLC of the Lactobacillus plantarum AF1 culture of the present invention according to an embodiment of the present invention. Figure 1a is a graph showing the fractions after the first injection of prep-HPLC, Figure 1b is a photograph showing the antifungal activity of the fraction (fraction) obtained through the first cycle after the first injection of prep-HPLC, Figure 1c is a photograph showing the antifungal activity of the fraction (fraction) obtained through the second cycle of the first injection.
Figure 2 is a second injection of the final fraction obtained by the first injection of the Lactobacillus plantarum AF1 culture solution of the present invention according to an embodiment of the present invention. Figure 2a is a graph showing the results after the second injection of prep-HPLC, Figure 2b is a fraction obtained through four cycles after the second injection of prep-HPLC, specifically through the second cycle Antifungal activity of the obtained fraction and the fraction obtained through the fourth cycle.
Figure 3 is a third injection of the final fraction obtained by the second injection of the Lactobacillus plantarum AF1 culture solution of the present invention according to an embodiment of the present invention. Figure 3a is a graph showing the fractions after the third injection of prep-HPLC, Figure 3b is a photograph showing the antifungal activity of the fraction (fraction) obtained through two cycles after the third injection of prep-HPLC.
Figure 4 is a fourth injection of the final fraction obtained by the third injection of the Lactobacillus plantarum AF1 culture solution of the present invention according to an embodiment of the present invention. Figure 4a is a graph showing the result of fractionation after the fourth injection of prep-HPLC, Figure 4b is a photograph showing the antifungal activity of the fraction (fraction) obtained through four cycles after the fourth injection of prep-HPLC.
Figure 5 is a photograph showing the results of the GC-MS analysis of the fraction finally confirmed as a fraction having antifungal activity through prep-HPLC according to an embodiment of the present invention.
Figure 6 is a photograph confirming the antifungal activity of the material of the two peaks subjected to GC-MS analysis according to an embodiment of the present invention, the left side is delta-dodecalactone (delta-dodecalactone), the right side is 9-octadi Ceramide (9-octadecenamide).
Figure 7 is a photograph showing the mass spectra (mass-spectrum) results of the substance and standard identified as a delta-dodecaractone separated from the supernatant of the Lactobacillus plantarum AF1 culture of the present invention according to an embodiment of the present invention As shown in FIG. 7A, the mass spectrum of the peak material of T 18.419 is measured by GC-MS analysis, and FIG. 7B is a mass spectrum result of a commercial standard of delta-dodecaractone.
Figure 8 is a photograph of the test results confirming the antifungal activity of the delta-dode caractone according to an embodiment of the present invention, each value means the amount of delta-dodecaractone used. 8A is Aspergillus It is a photograph showing the antifungal activity against fumigatus , Figure 8b is a photograph showing the antifungal activity against Candida albicans .

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

Example  One; Cultivation of Strains

To separate substances having antifungal activity and antibacterial activity in accordance with the present invention, Lactobacillus Planta room AF1 was cultured (Lb. Plantarum AF1), the culture of the Lactobacillus Planta room AF1 the (deMan Rogosa Sharpe) MSR broth (Difco Laboratories, USA) was carried out at 30 ℃ for 24 hours, and stored at -80 ℃ in MRS added 25% (v / v) glycerol.

Antifungal to confirm the antifungal activity of the isolated material was cultured under the following conditions.

First, Aspergillus fumigatus ATCC 96918, Aspergillus , purchased from the American Type Culture Collection (ATCC) flavus , ATCC 22546, Penicillium roqueforti ATCC 10110 and Aspergillus in the Lab petrakii PF-1, Aspergillus ochraceus PF-2, Aspergillus nidulans PF-3 and Cladosporium gossypiicola KF-2 was incubated for 3 days under conditions of 25 ° C. to 30 ° C. using malt-extracted whole medium (MEA, Difco Laboratories, USA) or PDA medium (potato dextrose agar, Difco Laboratories, USA). Stored at.

The fungi, specifically spores of the fungus, were recovered in order to carry out experiments to confirm antifungal activity. Recovery of the spores of the mold was carried out in the following manner. First, the fungi were incubated at 25 ° C. to 30 ° C. for 7 days until spores were produced in MEA medium or PDA medium. The resulting spores were extracted from the slope medium by shaking with sterile water containing Tween 80 (Tween 80, Polyoxyethyelene Sorbitan Monooleate) at 0.05% (v / v). The extracted spores were stored at −80 ° C. using a glycerol / water (20:80, v / v) solution (Hassan and Bullerman (2008)). After measuring the fungal spore density using a hemocytometer (manufactured by Marienfeld, Germany), the spores were stored at a concentration of 10 ⁷ spores per ml.

Candida albicans ATCC 24433 used sabouraud dextrose (SD) liquid medium (Difco Laboratories, USA) and Candida albicans ATCC 11006 and Candida albicans ATCC 18804 were incubated for 2 days at 25 ° C. to 37 ° C. using YM (Yeast mold) liquid medium (Difco Laboratories, USA).

In addition, the indicator strain for confirming the antimicrobial activity of the material isolated from the culture of the Lactobacillus plantarum AF1 was cultured under the following conditions.

Specifically, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 25922, Escherichia coli O157: H7 ATCC 43895, Micrococcus luteus ATCC 13513, Streptococcus mutans ATCC 25175, Pseudomonas aeruginosa ATCC 27853, Salmonella enterica serovar Typhi ATCC 19430 used LB (1% bacto-tryptone, 0.5% yeast extract, 1% NaCl) liquid medium and Listeria monocytogenes ATCC 19113 was used for brain heart infusion (BHI) liquid medium (Difco Laboratories, USA) and Staphylococcus aureus subsp. aureus ATCC 29213, and Enterococcus faecalis ATCC 29212, using a TS (tryptic soy) broth (Difco Laboratories, USA), and cultured overnight at a temperature of 37 ℃.

Example  2; Confirmation of antifungal activity

Antifungal activity was measured by the agar well diffusion method (Magnusson and Schnur (2001)) and the paper disc assay (Yang and Chang (2008)).

To measure the antifungal activity, plates were prepared by adding spores of A. fumigatus ATCC 96918 to 20 ml of MEA solid medium (1.5% agar, w / v) at a concentration of 10 ⁶ spores per 20 ml.

Specifically, in the case of the agar diffusion method, after forming 7.5 mm diameter holes in the prepared plate, 100 μl of samples were administered to each hole and left at room temperature for 1 hour before dispersing the administered samples into the medium. After the culturing step and the pre-culturing step was carried out by going through the culturing step of incubating at 30 ℃ for 48 hours. In the case of the paper disc method, 8 mm diameter paper discs (manufactured by Advantec Co., Japan) were placed on the prepared plate, 40 μl of sample was dropped on each paper disc, and then each plate was placed for 48 hours. Culture at 30 ° C., and measuring the formation of clear zones or inhibition zones (inhibition zones) was used.

Example  3; Isolation of substances with antifungal activity

Example  3-1. Preparation of Culture Supernatant and Analysis of Organic Acid

Lactobacillus plantarum AF1 of Example 1 was incubated for 24 hours at 30 ℃ using MRS liquid medium. After the incubation, the culture solution was centrifuged at 9500 × g for 15 minutes, and sterilized by 0.45 μm membrane filter (Advantec MFS, Inc. Japan) to prepare a culture supernatant.

Analysis of the organic acid of the prepared culture supernatant was performed using a Dionex-500 ion chromatography system (Dionex-500 ion chromatography system, Dionex, USA) equipped with an electro conductivity detector. Ion chromatography was performed using a Dionex IonPac ICE-AS6 column (9 x 253 mm) and at a flow rate of 1.0 ml / min at room temperature. Seven organic acids were detected by chemical suppressed conductivity using Anion-ICE micromembrane suppressor (Dionex, USA).

As a result of ion chromatography analysis, the highest amount of lactic acid is 21.21 mg / ml, oxalic acid is 1.48 mg / ml, citric acid is 0.7 mg / ml, and maleic acid is maleic. 0.483 mg / ml and acetic acid 0.379 mg / ml were detected. Tartaric acid was detected at 0.056 mg / ml and malic acid at 0.041 mg / ml.

Example  3-2. Determination of antifungal activity of culture supernatant

Antifungal activity was measured by performing a paper disk method on the organic acid detected from the culture supernatant.

Specifically, the paper disk method is 1.5% (w / v) B. aga (Bacto-agar, Netherlands) sensitive strain (sensitive mold) A. fumigatus ATCC 96918 or A. flavus , ATCC 22546 10 ⁶ spores per This was done using a plate prepared by addition at a concentration of 20 ml. The samples used for measuring the antifungal activity were prepared by concentrating the culture supernatant of the Lactobacillus plantarum AF1 and the organic acids detected from the culture supernatant 5 times, and the pH of each sample was adjusted to pH 3.9.

The antifungal activity results of the sample confirmed by the paper disc method are summarized in Table 1 below.

number Antifungal activity Sample name Clear zone size One O AF1 culture broth 15.74 ± 0.03 mm 2 O Oxalic acid (7.4 mg / ml) Less than 9.30 mm 3 X Maleic acid (2.415 mg / ml) - 4 O Acetic acid (1.895 mg / ml) Less than 9.30 mm 5 X Tartaric acid (0.28 mg / ml) - 6 O Lactic acid (106.05 mg / ml) 12.82 ± 0.02 mm 7 X Citric acid (3.5 mg / ml) - 8 X Malic acid (0.205 mg / ml) - 9 O Mixed solution of the organic acid component of the second to eighth 13.65 ± 0.04 mm

As shown in Table 1, the organic acid isolated from the culture supernatant of the Lactobacillus plantarum AF1 showed no antifungal activity (maleic acid, tartaric acid, citric acid and malic acid), or weak antifungal activity (oxalic acid and Acetic acid) and the concentrate of the culture supernatant of the Lactobacillus plantarum AF1 were found to have remarkably excellent antifungal activity. From the above results, antifungal activity of the culture supernatant of the Lactobacillus plantarum AF1 was expected to be due to a substance other than the organic acid contained in the culture supernatant.

Example  3-3. Of antifungal active substances Tablets1  - SPE ( Solid phase extraction ) refine

In order to purify antifungal substances from the culture supernatant of the Lactobacillus plantarum AF1, solid phase extraction (SPE) was first performed.

More specifically, the 24-hour culture supernatant of the Lactobacillus plantarum AF1 was isolated by SPE column (Isolute, C ₁₈ EC, 10 g; International Sorbent Technology Ltd., Hengoed, UK). The column was activated with 200 ml of methanol and stabilized with 200 ml of 10 mM sodium acetate, pH 4.0.

2.5 L of sample was passed per SPE column and the material adsorbed to the column was eluted with 30 ml of 95% aqueous acetonitrile. The eluted sample was concentrated using a vacuum centrifuge (Speed Vac, VS-802, Vison, Korea). Subsequent separation was carried out using prep-HPLC.

Example  3-4. Of antifungal active substances Tablets2  - HPLC  refine

The sample pretreated with SPE in Example 3-3 was UV detector (3702; Japan Analytical Industry Co., Ltd., Japan) and refractive index detector (RI-7; Japan Analytical Industry Co., Ltd., Japan) JAIGEL-GS (GFC) column (GS310; 13 μm, 20 × 500 mm, Japan Analytical Industry Co., Ltd, Japan) with an equipped recycling pre-HPLC system (LC9104; Japan Analytical Industry Co., Ltd., Japan) Purification was carried out using.

The antifungal activities of the samples were measured by the method of Example 2 using the agar diffusion method while monitoring using a UV detector (210 nm). The parts with antifungal activity were separated again using recycling prep-HPLC, and the process was repeated until the sample showing antifungal activity showed one pure peak.

More specifically, in order to isolate a substance having antifungal activity, a first stage HPLC and a second stage HPLC were performed using a JAIGEL-GS (GFC) column. As a result of performing the HPLC, the part having antifungal activity was purified again by performing the third step HPLC and the fourth step HPLC, and the JAIGEL-W (GPC) column ( W253 + W252; 20 x 500 mm, Japan Analytical Industry Co., Ltd., Japan). As an elution solvent, 80% (v / v) of water-soluble acetonitrile was used at a rate of 5 mL / min, and monitoring was performed at 210 nm using a UV detector.

The results of performing the first injection of prep-HPLC using the recycling prep-HPLC system are shown in FIG. 1.

As shown in FIG. 1, the first five kinds of fractions were obtained in the first injection of prep-HPLC, and fraction 2 (fraction ②) of the five kinds of fractions obtained in the first cycle of the first infusion was obtained. It was found to have the best antifungal activity (FIG. 1B). Four fractions were obtained by adding one cycle to fraction 2 having the best antifungal activity (fractions ②-1 to ②-4), and the second fraction of the fractions (②-2) It was confirmed that the most antifungal activity was excellent (Fig. 1c).

In the first injection, only the second fraction (②-2 fraction), which was found to have the best antifungal activity, was collected and a second injection was performed. A total of four cycles were performed at the second injection and the results are shown in FIG. 2.

As shown in FIG. 2, the antifungal activity of the fraction obtained in the second cycle (II-1 fraction) and the fraction obtained in the fourth cycle (II-2 fraction and II-3 fraction) through the second injection. It was confirmed that the antifungal activity was measured, the antifungal activity was confirmed in the II-1 fraction and II-2 fraction (Fig. 2b).

A third injection was performed by collecting the fractions (II-1 fraction and II-2 fraction) identified as having antifungal activity in the second infusion, and the results are shown in FIG. 3.

As shown in FIG. 3, a total of two cycles were performed (FIG. 3A), and a second fraction (III-2 fraction) obtained in the first cycle among the four fractions obtained as a result of the above performance had antifungal activity. Confirmed.

The fourth injection was performed by separating only the fraction (III-2) identified as having antifungal activity through the third injection, and the results are shown in FIG. 4.

As shown in FIG. 4, a total of four cycles were performed (FIG. 4A), and among the four fractions (IV-1 to IV-4 fractions) obtained as a result, the fraction obtained through the third cycle (IV). Fraction of -3) was found to have the highest antifungal activity.

The compound having antifungal activity was analyzed from the highest antifungal activity fraction (sample of IV-3) obtained through the fourth injection.

Example  3-4. Structural Analysis of Antifungal Active Substances

The process of analyzing the structure of the compound having antifungal activity from the fraction of the sample IV-3 obtained in Example 3-3 was performed by GC-MS analysis.

More specifically, GC-MS analysis was performed on a GC-MSD (HP 5890 gas chromatograph-5972 mass selective detector, Hewlett) equipped with a DB-5 ms column (30 m length × 0.25 mm id × 0.25 μm film thickness, J & W Scientific, USA). -Packard, USA). As a carrier gas, helium was used at a rate of 0.8 ml / min. The oven temperature was maintained at 100 ° C. for 1 minute and the temperature was raised to 250 ° C. at a rate of 5 ° C./min. The injector was kept at 200 ° C. and the detector temperature was kept at 250 ° C. The mass detector was operated in electron impact mode with an ionization energy of 70 eV. The results of conducting GC-MS analysis to identify the antifungal substance are shown in FIG. 5.

As shown in FIG. 5, two main peaks were detected and the peak of RT 18.419 was analyzed by delta-dodecalactone, and the peak of RT 31.459 was 9-octadecenamide. Was analyzed.

The antifungal activity of the two substances whose structure was finally confirmed was measured by the method of Example 2, and the results are shown in FIG. 6.

The disk on the left side of FIG. 6 includes delta-dodecaractone, and the disk on the right side contains 9-octadecenamid. As shown in FIG. 6, delta-dodecaractone showed strong antifungal activity while 9-octadecenamid was confirmed to be a substance without antifungal activity.

Thus, the identified antifungal substance was found to be delta-dodecaractone.

Example  3-5. Confirmation of the structure of the antifungal active substance

In order to confirm the structure of the material searched at the peak of RT 18.419 identified as delta-dodecaractone in Example 3-4, the material searched at the peak of RT 18.419 and commercially available delta-dodecaractone were used as standards. Its mass-spectrum was compared and the results are shown in FIG. 7. 7A is a mass spectral result of the material retrieved at the peak of RT 18.419, and FIG. 7B is a mass spectral result of the commercial delta-dodecaractone.

As shown in FIG. 7, the peak of RT 18.419 and the commercially available delta-dodecalactone exhibit the same mass-spectrum so that the antifungal substance identified in the present invention finally has a delta. It was found to be delta-dodecalactone.

Example  4 deltas Dodekaractone  Antifungal Spectrum and Antimicrobial Spectrum Identification

4-1. delta- Dodekaractone  Identification of antifungal spectra

In order to confirm the antifungal spectrum of the delta-dodecalactone (MIC), a minimal inhibition concentration (MIC) was measured by spot-on-the lawn assay for the fungi of Example 1.

Preparation of a sample for performing the spot-on-the lawn assay method after diluting 2 mg of the commercially available delta-dodecaractone having the same structure as the separated delta-dodecaractone in 100 μl of 100% methanol, Dilution was sequentially performed twice in 10 mM phosphate buffer to prepare samples for each concentration.

The plate of indicator strain for performing the spot-on-the lawn assay method was prepared by adding 10 ⁶ cfu / 20 ml (spore / medium) of each fungal spore to MEA medium or PDA medium in case of mold. For yeast, Candida albicans strains were prepared by smearing at a concentration of 10 ⁶ cfu / plate on SDA medium or YMA medium.

A spot-on-the lawn assay was performed to drop 10 μl of the sample for each concentration of delta-dodecaractone on the prepared medium. The delta-dodecaractone treated medium was cultured at 25 or 30 ° C. for 48 hours to 72 hours, and MIC was performed by measuring a minimum concentration showing growth inhibitory activity, and the results are shown in Table 2 and FIG. 8. Shown in 8A is Aspergillus MIC results for fumigatus ATCC 96918, FIG. 8B shows Candida albicans MIC result for ATCC 24433.

Indicative strain MIC (ug) Aspergillus flavus , ATCC 22546 6.25 Aspergillus fumigatus ATCC 96918 6.25 Aspergillus petrakii PF-1 6.25 Aspergillus ochraceus PF-2 6.25 Aspergillus nidulans PF-3 6.25 Cladosporium gossypiicola KF-2 12.50 Penicillium roqueforti ATCC 10110 6.25 Candida albicans ATCC 24433 12.50 Candida albicans ATCC 11006 12.50 Candida albicans ATCC 18804 12.50

As shown in Table 2 and FIG. 8, the delta-dodecaractone was confirmed to exhibit antifungal activity against both the indicator strains (FIG. 8a) and yeast (FIG. 8b), and compared to candida albicans. It was confirmed that the antifungal activity against.

4-2. delta- Dodekaractone  Confirmation of Antimicrobial Spectrum

The antibacterial spectrum of the delta-dodecalactone was measured for the bacteria of Example 1 using the paper disc method of Example 2.

In order to perform the paper disc method, 500 μg of the commercially available delta-dodecaractone having the same structure as the separated delta-dodecaractone was dissolved in 40 μl of 60% methanol to prepare a sample.

Plates of the indicator strain for performing the paper disc method was prepared using the bacteria of Example 1. Specifically, Bacillus among the bacteria subtilis ATCC 6633, Escherichia coli ATCC 25922, Escherichia coli O157: H7 ATCC 43895, Micrococcus luteus ATCC 13513, Streptococcus mutans ATCC 25175, Pseudomonas aeruginosa ATCC 27853, Salmonella enterica Plates for confirming antimicrobial activity against serovar Typhi ATCC 19430 were prepared by smearing at a concentration of 10 ⁶ cfu / plate in LB solid medium, Listeria Plates for confirming antimicrobial activity against monocytogenes ATCC 19113 were prepared by smearing BHI solid medium at a concentration of 10 ⁶ cfu / plate, Staphylococcus aureus subsp . aureus ATCC 29213 and Enterococcus Plates to confirm the antimicrobial activity against faecalis ATCC 29212 was prepared by plating at a concentration of 10 ⁶ cfu / plate in TSA medium.

In the paper disc method, a delta-dodecaractone sample (500 μg / 40 μl 60% methanol) was prepared by placing a 8 mm diameter paper disc on a plate medium coated with the indicator strain and incubated overnight at 37 ° C., followed by each indication. The growth inhibition activity of the strain was measured, and the results are shown in Table 3 below.

Indicative strain Antimicrobial activity Bacillus subtilis ATCC 6633 + Micrococcus luteus ATCC 13513 - Streptococcus mutans ATCC 25175 + Staphylococcus aureus subsp . aureus ATCC 29213 + Enterococcus faecalis ATCC 29212 + Listeria monocytogenes ATCC 19113 + Escherichia coli ATCC 25922 + Escherichia coli O157: H7 ATCC 43895 - Pseudomonas aeruginosa ATCC 27853 - Salmonella enterica serovar Typhi ATCC 19430 -

As shown in Table 3, the delta-dodecaractone was confirmed to have antibacterial activity against various bacteria. Specifically, the delta-dodecaractone has been shown to have antimicrobial activity against Bacillus subtilis, Streptococcus mutans, Staphylococcus aureus, Enterococcus pecalis, Listeria monocytogenes and Escherichia coli, against antimicrobial Gram-positive bacteria The spectrum was found to be wider than the antibacterial spectrum for Gram-negative bacteria.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concepts of the present invention defined in the following claims are also provided. It belongs to the scope of rights.

Claims (8)

Compound having antifungal activity and antibacterial activity represented by the following formula (1).
[Formula 1]

An antifungal composition comprising the compound represented by the following formula (1) as an active ingredient.
[Formula 1]

The method of claim 2, wherein the composition is Aspergillus sp., Cladosporium sp., Penicillium sp. And Candida sp. An antifungal composition having antifungal activity against at least one fungus selected from the group consisting of: The method of claim 2, wherein the composition is Bacillus bacteria (Bacillus sp.), Streptococcus bacteria (Streptococcus sp.), Stearyl Philo Cocos bacteria (Staphylococcus sp.), Enterococcus bacteria (Enterococcus sp.), Listeria monocytogenes (Listeria sp .) And E. coli ( E. coli ) antifungal composition, characterized in that it further has an antimicrobial activity against one or more bacteria selected from the group consisting of A preservative comprising the composition according to claim 2 as an active ingredient. Cosmetic composition comprising the composition according to claim 2 as an active ingredient. A composition for the treatment or prevention of fungal diseases comprising the composition according to claim 2 as an active ingredient. According to claim 7, wherein the fungal disease is fungal dermatitis, athlete's foot and fungal pneumonia composition for the treatment or prevention of fungal diseases of at least one selected from the group consisting of.

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