Antifungal composition comprising senna tora sprout extract

Abstract

The present invention relates to an antifungal composition comprising a Senna tora sprout extract or a fraction thereof as an active ingredient, wherein the Senna tora sprout extract or a fraction thereof or a compound isolated therefrom exhibits a photoreaction under light conditions of a wavelength of 400-528 nm. The antifungal composition of the present invention has a synergistic effect on antifungal activity upon irradiation with light having a wavelength of 400-528 nm.

Description

Antifungal composition comprising cassia seed sprout extract

[0001] The present invention relates to an antifungal composition comprising an extract of Cassia tora sprouts or a fraction thereof as an active ingredient.

[0002]

[0003] Trichophyton causes dermatophytosis in humans and animals and can invade any part of the body from head to toe. Fungal skin diseases are classified into deep dermatophytosis and superficial dermatophytosis, and tinea is one of the superficial dermatophytosis, and the fungus that parasitizes the outermost layer of the skin is called trychophyton. Depending on the area of ​​skin invasion, if it occurs on the hands and feet, it is called tinea manus tinea pedis, if it occurs on the face, it is called tinea faciale, if it occurs on the head, it is called tinea capitis, if it occurs in the groin, it is called tinea crusis, if it occurs on the fingernails and toenails, it is called tinea ungium or onychomycosis, and if it occurs in any other area, it is called tinea corporis. The location of occurrence and characteristics of the lesions of tinea versicolor are determined by the type of dermatophyte that is transmitted.

[0004] The most common dermatophyte infection is tinea pedis (athlete's foot), which occurs between the toes, particularly between the fourth and fifth toes or between the third and fourth. The skin between the toes typically becomes irritated, swollen, whitish, cracked, and scaly. Excessive sweating can lead to unpleasant foot odor and sometimes itching. Meanwhile, intensely itchy blisters may form on the soles or edges of the feet. In some cases, the entire soles of the feet may thicken and flake off in a fine, powdery layer without itching.

[0005] Not only humans, but also plants are infected with various fungi. Aspergillus flavus, which produces aflatoxin, a carcinogen in various grains, Rhizoctonia solani, which causes sheath blight, Colletotrichum gloeosporioides, which causes anthracnose in persimmons and peppers, and Botrytis cinerea, which causes gray mold disease in asparagus, are all examples of fungi that expose plants to disease.

[0006]

[0007] In the present invention, an attempt was made to develop a composition having antifungal activity against fungi that cause various diseases not only in humans but also in plants.

[0008]

[0009] The present inventors have made great efforts to find a natural product that suppresses the activity of fungi that infect humans and plants, while having few side effects and is harmless to the human body. As a result, they have confirmed that when an extract of Cassia tora sprouts or a fraction thereof or a compound isolated therefrom is irradiated with LED light, the antifungal activity against the fungus that causes ringworm and the antifungal activity against plant-infecting fungi are enhanced / amplified, thereby completing the present invention.

[0010]

[0011] In order to achieve the above-mentioned purpose, the object of the present invention is to provide an antifungal composition comprising a cassia seed sprout extract or a fraction thereof as an active ingredient.

[0012] Another object of the present invention is to provide a method for preventing or treating tinea, comprising the steps of applying the antifungal composition to the skin or nails of an individual; and the steps of irradiating the skin or nails of an individual with light having a wavelength of 400 nm to 528 nm.

[0013] Another object of the present invention is to provide a method for isolating a compound having antifungal activity.

[0014] Another object of the present invention is to provide an antifungal method comprising a step of administering, applying or treating an extract of Cassia tora sprouts or a fraction thereof or an antifungal composition comprising the same to a subject or fungus.

[0015] Another object of the present invention is to provide a use of a cassia seed sprout extract or a fraction thereof for the manufacture of an antifungal or anti-ringworm medicine or food.

[0016]

[0017] The present invention relates to an antifungal composition comprising a Cassia tora sprout extract or a fraction thereof as an active ingredient, wherein the Cassia tora sprout extract or a fraction thereof or a compound isolated therefrom exhibits a photoreaction under light conditions having a wavelength of 400 to 528 nm.

[0018] The cassia seed sprout extract of the present invention or a fraction thereof or a compound isolated therefrom exhibits a synergistic effect of antifungal activity when irradiated with light having a wavelength of 400 nm to 528 nm.

[0019]

[0020] Figure 1 shows HPLC-DAD (High-performance liquid chromatography with diode-array detection) of the dichloromethane fraction (MC fr.) of Cassia tora sprouts.

[0021]

[0022] The following describes this specification in more detail.

[0023] Each description and embodiment disclosed in this invention can be applied to other descriptions and embodiments thereof. That is, all combinations of the various elements disclosed in this invention fall within the scope of this invention. Furthermore, the scope of this invention is not limited by the specific descriptions described below.

[0024] Expressions such as “comprising” as used herein should be understood as open-ended terms implying the possibility of including other embodiments, unless specifically stated otherwise in the phrase or sentence in which the expression is included.

[0025] The terms or words used in the description and claims of the present invention should not be interpreted as limited to their usual or dictionary meanings, but should be interpreted as meanings and concepts that conform to the technical idea of ​​the present invention, based on the principle that the inventor can appropriately define the concept of the term to explain his or her own invention in the best way.

[0026]

[0027] As one embodiment to achieve the above purpose, the present invention provides an antifungal composition comprising a cassia seed sprout extract or a fraction thereof as an active ingredient.

[0028] The antifungal composition of the present invention comprises cassia seed sprouts.

[0029] In the present invention, the term "cassia tora sprout" refers to a stage from the soaking stage of cassia tora seeds to the germination stage, the stage in which cotyledons grow, and the stage before true leaves emerge. Specifically, the cassia tora sprout refers to a seed before germination after soaking in water, a germinated seed, and a germinated cotyledon. That is, it refers to a state in which cassia tora seeds are soaked in water, cassia tora seeds sprouted after soaking, and young sprouts grown by sowing cassia tora seeds after soaking, and before true leaves emerge.

[0030] The above cassia seedlings are artificially cultivated in indoor farm facilities such as plant factories, and can be easily cultivated indoors. They can be cultivated and used in a short period of time, such as 1 to 10 days, without separate nutrient supply, so they have the advantage of great industrial usability.

[0031] The above "cassia seed" is the seed of cassia tea and is called cassia seed. The seed is columnar, 3-6 mm long and 2-3.5 mm in diameter, with hairs, and has light yellow-brown vertical lines and stripes on both sides.

[0032] The above-mentioned cassia seed sprouts include soaked cassia seeds, germinated cassia seeds, and sprouts from which cotyledons have grown. The cassia seed sprouts from which cotyledons have grown may include whole plants, above-ground parts, or underground parts. The above-mentioned whole plants include roots, stems, and leaves. The above-mentioned above-ground parts include stems and leaves, and the above-mentioned underground parts refer to roots.

[0033] In the present invention, the term "extract" includes an extract obtained by extraction treatment, a diluted or concentrated extract, a dried product obtained by drying the extract, a controlled or purified product of the extract, or a mixture thereof, the extract itself, and all formulations that can be formed using the extract.

[0034] In the present invention, drying of the extract can be performed using known methods, as long as the useful components of the collected plant are not destroyed. For example, drying can be performed naturally in a shaded area. Furthermore, the crushing or pulverizing can be performed to a degree that allows sufficient extraction of the plant's useful components during the subsequent extraction process. The drying and crushing or pulverizing processes can be performed in reverse order or repeated as needed.

[0035] In the extraction of the present invention, the extraction method is not particularly limited, and extraction can be performed according to a method commonly used in the relevant technical field. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, reflux extraction, and cold immersion extraction, and these may be performed alone or in combination of two or more methods.

[0036] In the extraction of the present invention, the extraction method is not particularly limited, and extraction can be performed according to a method commonly used in the relevant technical field.

[0037] In addition, in the present invention, the extract can be obtained by extracting the cassia tora sprouts with water, a C1 to C4 alcohol, or a mixed solvent thereof. In addition, the alcohol can be ethanol or methanol, and the ethanol can be 30 to 100% ethanol, and the methanol can be 30 to 100% methanol. Specifically, the extract of the cassia tora sprouts can be an ethanol extract of the cassia tora sprouts.

[0038] The term "fraction" used in the present invention means a result obtained by performing fractionation to separate a specific component or a specific group of components from a mixture containing various components.

[0039] The above fraction may be a fraction obtained by applying a method selected from the group consisting of solvent fractionation, ultrafiltration fractionation, chromatography fractionation, and combinations thereof.

[0040] In the present invention, the fraction can be interpreted as a fraction containing aurantio-optusin, emodin, or chrysophanol, obtained by applying the cassia seed sprout extract or its fractions to various fractionation methods. The fraction of the extract can be a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, a chromatographic fractionation method performed by performing various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity), etc. In particular, the solvent used in the solvent fractionation method is not particularly limited thereto, but a polar solvent or a non-polar solvent can be used, and preferably a non-polar solvent can be used.

[0041] The fractionation method for obtaining the solvent fraction in the present invention is not particularly limited and may be performed according to methods commonly used in the art. A non-limiting example of the fractionation method includes a method of treating a cassia seedling extract with a predetermined solvent to obtain a fraction from the extract.

[0042] In the present invention, the type of solvent used to obtain the solvent fraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvent include water, alcohols having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate, or mixed solvents thereof.

[0043] The solvent fraction can be prepared by fractionating the Cassia tora sprout extract with water, an alcohol having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate, or a mixed solvent thereof. Specifically, the solvent fraction can be a fraction of the Cassia tora sprout ethanol extract with hexane, dichloromethane, ethyl acetate, butanol, water, or a mixed solvent thereof.

[0044] In one embodiment of the present invention, Cassia tora sprouts were extracted with ethanol at 1 to 100 times (w / v) to obtain an ethanol extract of Cassia tora sprouts, which was sequentially fractionated in the order of n-hexane (Hex), dichloromethane (Dichloromethane), ethyl acetate (EtOAc), n-butanol (BuOH), and water (H2O), thereby producing a total of five solvent fraction layers: normal hexane, dichloromethane, ethyl acetate, normal butanol, and a residual water layer.

[0045] In the present invention, a compound can be separated from the fraction by combining methods such as solvent fractionation, ultrafiltration fractionation, and chromatography fractionation, and the compound separated from the fraction can be one or more compounds selected from the group consisting of aurantio-optusin, emodin, and chrysophanol.

[0046] In the present specification, the cassia seed sprout extract or fraction thereof exhibits photoreactivity under light conditions having a wavelength of 400 nm to 528 nm.

[0047] In the present invention, the term "antifungal composition exhibiting photoreactivity" means an antifungal composition that is used together with a medical device capable of irradiating light under specific conditions using an LED / OLED light source or the like, and whose antifungal activity increases in response to light under specific conditions.

[0048] The antifungal composition of the present invention comprises the Cassia tora sprout extract or a fraction thereof, and the Cassia tora sprout extract or a fraction thereof exhibits photoreactivity and maximum activity under light conditions having a wavelength of 400 nm to 528 nm. Specifically, when the Cassia tora sprout extract or a fraction thereof is irradiated with light having a wavelength of 400 nm to 528 nm together with the Cassia tora sprout extract or a fraction thereof, the Cassia tora sprout extract or a fraction thereof exhibits photoreactivity, while the antifungal activity is enhanced. Since the Cassia tora sprout extract or a fraction thereof exhibits photoreactivity when irradiated with light having a wavelength of 400 nm to 528 nm, when treated together with light having a wavelength of 400 nm to 528 nm, the antifungal activity against Trichophyton (Athlete's foot) or plant disease-causing fungi is enhanced compared to when the Cassia tora sprout extract or a fraction thereof is treated alone. Specifically, the 455±2 nm LED light used in the embodiment of the present invention has a lambda (λ) peak wavelength of 455±2 nm and a wide wavelength range of 400 nm to 528 nm when measured with a spectrophotometer.

[0049] The aurantio-optusin, emodin, and chrysophanol compounds separated from the above fractions also exhibit photoreactivity under light conditions with a wavelength of 400 nm to 528 nm, and therefore, when the compounds are irradiated together with light with a wavelength of 400 nm to 528 nm, the antifungal activity is enhanced compared to when the compounds are treated alone.

[0050] The luminous intensity of the above light is 10 to 2,000 μmol m -2 s -1, and light with a wavelength of 400 nm to 528 nm can be irradiated together with the Cassia tora sprout extract from a distance of 1 to 30 cm. In addition, the light can be irradiated more than once for 5 to 20 minutes, specifically, 1 to 6 times a day for 5 to 20 minutes, and irradiation is possible for 3 days to 6 months. When the light irradiation time is 10 minutes or more, specifically 10 to 20 minutes, rather than less than 10 minutes, the Cassia tora sprout extract or its fraction exhibits maximum activity against light, thereby increasing antifungal activity.

[0051] The term "irradiation" in the present invention means shining light on a surface, that is, exposing the skin or fingernails to which the cassia seed sprout extract or a fraction thereof has been applied, to light from an artificial light source.

[0052] Additionally, in the present invention, the term “active ingredient” means an ingredient that exhibits the desired activity alone or can exhibit the activity together with a carrier that is inactive in itself.

[0053] The term "antifungal" in the present invention may refer to bacteria, specifically fungi, that cause diseases in humans or plants. More specifically, it refers to inhibiting or killing the growth of trichophyton or plant disease-causing bacteria.

[0054] Specifically, it may have antifungal activity against trichophyton, eflatoxin-producing bacteria or sheath blight bacteria. The trichophyton may be selected from the group consisting of Epidermophyton floccosum, Trichophyton interdigitale, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton verrucosum and Trichophyton violaceum.

[0055] Additionally, the eflatoxin-producing bacteria may be Aspergillus flavus, and the leaf sheath blight bacteria may be Rhizoctonia solani.

[0056] The above antifungal composition is used together with a skin care device that suppresses or improves the activity of tinea versicolor (tinea fungi) by irradiating the skin or fingernails or toenails with light under specific conditions using an LED light source or the like, and refers to an antifungal composition whose antifungal activity increases in response to light under specific conditions.

[0057] The light may be light derived from an LED or an OLED. In the present invention, the light may utilize various types of light-emitting means such as a resistive bulb, a discharge lamp, a light-emitting diode (LED), an organic light-emitting diode (OLED), and specifically, the light may be light derived from a light-emitting diode (LED) or an organic light-emitting diode (OLED).

[0058] The above antifungal composition can be applied (used) as one or more selected from the group consisting of a pharmaceutical composition, a food composition, a food additive composition, a quasi-drug composition, a cosmetic composition, and a skin external application composition.

[0059] Specifically, the antifungal composition may be applied (used) as one or more selected from the group consisting of a pharmaceutical composition for treating or preventing tinea, a food composition for improving or preventing tinea, a food additive composition for improving or preventing tinea, an over-the-counter drug composition for improving or preventing tinea, a cosmetic composition for improving or preventing tinea, and a skin external application composition for treating or preventing tinea.

[0060] The above antifungal composition can be applied as a pharmaceutical composition for the treatment or prevention of tinea. The pharmaceutical composition of the present invention can be administered orally or parenterally during clinical administration, but is typically administered topically directly to the skin in the form of an external preparation.

[0061] In addition, when formulating the pharmaceutical composition of the present invention, it can be formulated using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants that are commonly used.

[0062] Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing the active ingredient of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, and gelatin. In addition to simple excipients, lubricants may also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, may be included. Preparations for parenteral administration include topical preparations such as creams, lotions, ointments (semi-solid external preparations), microemulsions, gels, pastes, and transdermal preparations (TTS).

[0063] The pharmaceutical composition of the present invention is typically administered at a daily dose of 0.001 to 150 mg / kg body weight, and may be administered once or in several divided doses. However, the actual dosage of the pharmaceutical composition of the present invention is determined in light of various related factors such as the route of administration, the patient's age, sex, body weight, and the patient's severity of condition, and therefore the above dosage should not be construed as limiting the scope of the present invention in any way.

[0064] The above antifungal composition can be applied as a food composition for improving or preventing tinea. The food composition of the present invention can include forms such as pills, powders, granules, infusions, tablets, capsules, or liquids, and there is no particular limitation on the types of foods to which the cassia seed sprout extract of the present invention or its fractions can be added. Examples of foods to which the above substance can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

[0065] The above food composition may contain other ingredients in addition to the cassia seed sprout extract, its fractions, or compounds isolated therefrom, and the types thereof are not particularly limited. For example, as with conventional foods, it may contain various herbal extracts, food science-acceptable food additives, or natural carbohydrates as additional ingredients, but is not limited thereto.

[0066] In the present invention, the term "food supplement additive" means a component that can be added to food as an auxiliary, and can be appropriately selected and used by those skilled in the art as added in the production of food of each formulation. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., but the types of food supplement additives of the present invention are not limited by the above examples.

[0067] Examples of the above natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. In addition to the above, natural flavoring agents (thaumatin, etc.), stevia extracts (rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used as flavoring agents.

[0068] The food composition of the present invention may include a health functional food. The term "health functional food" as used herein refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients having functionality useful to the human body. Here, functionality means obtaining a beneficial effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological functions. The health functional food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, raw materials and ingredients commonly added in the art can be added. In addition, unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it uses food as a raw material, and can be highly portable.

[0069] The amount of active ingredients mixed can be appropriately determined depending on the intended use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food, the active ingredient of the present invention can be added in an amount of 0.01 to 50 wt%, preferably 0.1 to 10 wt%, of the raw material composition, but is not limited thereto. However, in the case of long-term intake for the purpose of health and hygiene or health regulation, the amount may be used below the above range.

[0070] The above antifungal composition can be used as a food additive composition for improving or preventing tinea versicolor. When used as a food additive, the composition can be added as is or combined with other foods or food ingredients, and can be used appropriately according to conventional methods.

[0071] The amount of active ingredients mixed can be appropriately determined depending on the intended use. Generally, the Cassia tora sprout extract of the present invention, or a fraction thereof, or a compound isolated therefrom can be included in an amount of from about 0.01 wt % to about 99.99 wt % of the total weight, preferably from about 0.1 wt % to about 99 wt % of the weight of the composition.

[0072] The antifungal composition of the present invention can be applied as a cosmetic composition for improving or preventing tinea, and when applied as an antifungal cosmetic composition, in addition to the effective ingredient, cassia seed sprout extract or a fraction thereof or a compound isolated therefrom, it can include a compound or natural extract known to have an antifungal effect so as to increase or enhance the antifungal effect.

[0073] Meanwhile, in the antifungal cosmetic composition of the present invention, the effective ingredient may be included in any amount (effective amount) depending on the intended use, formulation, mixing purpose, etc., as long as it can exhibit antifungal activity. Typically, the effective amount may be included within the range of 0.001 wt % to 99.99 wt % based on the total weight of the composition. Here, the "effective amount" refers to the amount of the effective ingredient capable of inducing an antifungal effect. This effective amount can be experimentally determined within the normal ability of a person skilled in the art.

[0074] The antifungal cosmetic composition of the present invention can be manufactured in various forms. For example, the cosmetic composition of the present invention can be manufactured in any formulation commonly manufactured in the art, and can be formulated as, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray, but is not limited thereto. In addition, specifically, it may have a formulation selected from the group consisting of skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, soap, shampoo, rinse, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, milk, lipstick, makeup base, foundation, pressed powder, and loose powder, but is not limited thereto.

[0075] The antifungal cosmetic composition of the present invention may include, in addition to its active ingredient, a carrier acceptable for use in cosmetic formulations. Herein, the term "acceptable carrier for use in cosmetic formulations" refers to a compound or composition already known and used, or a compound or composition to be developed in the future, that can be included in cosmetic formulations and that does not exhibit toxicity, instability, or irritation beyond what the human body can tolerate when in contact with the skin. The carrier may be included in the antifungal cosmetic composition of the present invention in an amount of from about 0.01 wt % to about 99.99 wt % of the total weight of the composition, preferably from about 0.1 wt % to about 99 wt % of the composition's weight. However, since the above ratio varies depending on the formulation of the cosmetic of the present invention, its specific application site (face or hands), its preferred application amount, etc., the above ratio should not be construed as limiting the scope of the present invention in any way.

[0076] Meanwhile, examples of the carrier include alcohol, oil, surfactant, fatty acid, silicone oil, humectant, moisturizer, viscosity modifier, emulsion, stabilizer, UV blocker, colorant, fragrance, etc. Compounds / compositions that can be used as the carrier, such as alcohol, oil, surfactant, fatty acid, silicone oil, humectant, moisturizer, viscosity modifier, emulsion, stabilizer, UV blocker, colorant, fragrance, etc., are already known in the art, so a person skilled in the art can select and use an appropriate corresponding substance / composition.

[0077] The above antifungal composition can be applied as an over-the-counter drug composition for improving or preventing tinea. When the composition of the present invention is used as an over-the-counter drug additive, the Cassia tora sprout extract of the present invention, its fractions, or compounds isolated therefrom can be added as is or used in combination with other over-the-counter drugs or over-the-counter drug ingredients, and can be used appropriately according to conventional methods. The amount of active ingredient mixed can be appropriately determined depending on the intended use.

[0078] The term "quasi-drug" used in the present invention refers to products that are used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals, and have a milder effect than pharmaceutical products. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products other than those used for pharmaceutical purposes, and include products used for treating or preventing diseases of humans or animals, products that have a mild effect on the human body or do not directly affect it.

[0079] The pharmaceutical composition of the present invention is not limited thereto, but may preferably be a disinfectant, shower foam, wet tissue, detergent soap, hand wash, ointment, or patch.

[0080] The above antifungal composition can be applied as an external composition for treating or preventing tinea.

[0081] The term "external preparation" of the present invention refers to a preparation provided for external use, and includes external powders, external tablets, external solutions, ointments, ointments, suppositories, etc., and the skin external preparation of the present invention includes, without limitation, preparations that act specifically on the skin externally.

[0082] The topical skin preparation according to the present invention may be a parenteral administration preparation formulated in a solid, semi-solid, or liquid form by adding commercially available inorganic or organic carriers, excipients, and diluents. The preparation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions, and emulsions, but is not limited thereto.

[0083] Carriers, excipients and diluents that may be included in the above external preparations include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0084] In the external skin composition according to each formulation, a person skilled in the art can appropriately select and mix other ingredients other than the cassia seed sprout extract of the present invention or a fraction thereof or a compound isolated therefrom, according to the formulation or purpose of use of other external skin compositions, and in this case, a synergistic effect may occur when applied simultaneously with other raw materials.

[0085] The skin external application composition of the present invention may contain 0.0001 to 30% (w / v) of the effective ingredient relative to the total weight of the composition.

[0086]

[0087] In another aspect, the present invention provides a method for preventing or treating tinea, comprising the steps of applying the antifungal composition to the skin or nails of a subject; and irradiating the skin or nails of the subject with light having a wavelength of 400 nm to 528 nm.

[0088] In the present invention, the antifungal composition, prevention and treatment of tinea are as described above.

[0089] As used herein, the term "application" refers to contacting the antifungal composition with the skin or nails of an individual using any suitable method, thereby causing the components of the composition to act on Trichophyton to exert a bactericidal effect. The dosage of the antifungal composition may be appropriately determined depending on the intended use.

[0090] The subject to which the antifungal composition of the present invention is applied includes, without limitation, mammals including humans, but examples thereof may include cows, pigs, horses, rabbits, rats, and humans.

[0091] In addition, the amount of the antifungal composition of the present invention can be appropriately adjusted according to individual differences such as age, degree of lesion, etc., and formulation, and can be used for one week to several months by applying an appropriate amount to the skin or fingernails once or several times a day.

[0092] After applying the above antifungal composition, light having a wavelength of 400 nm to 528 nm can be irradiated to the skin or fingernails to which the Cassia tora sprout extract or a fraction thereof or a compound isolated therefrom has been applied, from a distance of 1 to 30 cm. In addition, the light can be irradiated at least once for 5 to 20 minutes, and specifically, the light can be irradiated at least once a day for 5 to 20 minutes, and specifically, the light can be irradiated 1 to 6 times a day for 5 to 20 minutes, and the light can be irradiated for 3 days to 6 months.

[0093]

[0094] In another aspect, the present invention provides a method for isolating a compound having antifungal activity, comprising: a first step of preparing a Cassia tora sprout extract; a second step of fractionating the Cassia tora sprout extract with water, an alcohol having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate or a mixed solvent thereof to prepare a fraction of the Cassia tora sprout extract; and a third step of isolating at least one compound selected from the group consisting of aurantio-optucin, emodin and chrysophanol from the fraction of the Cassia tora sprout extract.

[0095] The method for isolating a compound having antifungal activity of the present invention comprises the first step of preparing a Cassia tora sprout extract. As described above, the extract can be prepared by extracting Cassia tora sprouts with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

[0096] The method for isolating a compound having antifungal activity of the present invention includes two steps of preparing a fraction of the Cassia tora sprout extract by fractionating the Cassia tora sprout extract with water, an alcohol having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate, or a mixed solvent thereof.

[0097] As described above, the Cassia tora sprout extract prepared in step 1 can be fractionated with water, an alcohol having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate, or a mixed solvent thereof to prepare a solvent fraction of the Cassia tora sprout extract. Specifically, a dichloromethane fraction of the Cassia tora sprout ethanol extract can be prepared.

[0098] The method for isolating a compound having antifungal activity of the present invention includes a third step of isolating at least one compound selected from the group consisting of aurantio-optucin, emodin, and chrysophanol from the solvent fraction of the cassia seedling extract prepared in the second step.

[0099] As described above, the aurantio-optusin, emodin and chrysophanol are contained in the Cassia tora sprout extract or its solvent fraction and can therefore be separated from these extracts or fractions.

[0100] As described above, the solvent fraction of the cassia seed sprout extract prepared in the above 2 steps can be separated into aurantio-optucin, emodin, and chrysophanol by performing chromatographic fractionation (HPLC or / and MPLC).

[0101] The compound having antifungal activity of the present invention can be separated by applying a method selected from the group consisting of solvent fractionation, ultrafiltration fractionation, chromatography fractionation, and combinations thereof, and the fraction can be interpreted as a fraction containing aurantio-optusin, emodin, or chrysophanol obtained by applying a combination of various fractionation methods to the cassia seedling extract or fractions thereof.

[0102]

[0103] In another aspect, the present invention provides an antifungal method comprising a step of administering, applying or treating an object or fungus with an extract of Cassia tora sprouts or a fraction thereof or an antifungal composition comprising the same.

[0104]

[0105] In another aspect, the present invention provides the use of a cassia seed sprout extract or a fraction thereof for the manufacture of an antifungal or anti-ringworm medicine or food.

[0106] In the present invention, the terms “cassia seed sprout extract”, “fraction”, “antifungal composition”, “ringworm”, “antifungal”, “application”, and “entity” are as described above.

[0107] The term "administration" in the present invention means introducing a predetermined substance into an individual by an appropriate method, and the route of administration of the Cassia tora sprout extract or a fraction thereof or an antifungal composition comprising the same may be administered via any common route as long as it can reach the target tissue. Examples of such administration include, but are not limited to, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration.

[0108] The term "treatment" in the present invention can be appropriately applied according to conventional methods well known to those skilled in the art in various fields requiring antifungal agents. The treatment amount of the Cassia tora sprout extract or its fractions or an antifungal composition containing the same can be appropriately determined depending on the intended use.

[0109]

[0110] Matters mentioned in the antifungal composition of the present invention or the formulation, treatment method, antifungal method and separation method including these compositions are equally applicable unless they are contradictory to each other.

[0111]

[0112] Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings so that those skilled in the art can easily implement the present invention. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0113]

[0114] [Preparing experimental materials]

[0115] 1. Preparation of plant materials

[0116] Cassia tora sprouts and seeds grown in an indoor farm were used. The cassia tora sprouts include seeds that have been soaked in water before germination, sprouted seeds, and sprouted cotyledons, and are in a state before true leaves emerge.

[0117]

[0118] 2. LED light irradiation

[0119] The LED untreated group was not treated with LED light, and the LED treated group was treated with LED light with a lambda (λ) peak wavelength of 455±2 nm for 15 minutes per day (15 minutes / day) for 2 days.

[0120]

[0121] 3. Strains for antifungal activity testing

[0122] The strains in Table 1 were used as strains for antifungal activity testing.

[0123] No. Genus Name Species Name Symptoms Athlete's foot fungus 1 Epidermophyton floccosum Toenail fungus 2 Trichophyton interdigitale Athlete's foot 3 Trichophyton mentagrophytes Athlete's foot 4 Trichophyton rubrum Athlete's foot 5 Trichophyton verrucosum Athlete's foot 6 Trichophyton violaceum Athlete's foot Human disease-causing agent 1 Aspergillus fumigatus Aspergillosis 2 Candida albicans Candida vaginitis Plant disease-causing agent 1 Aspergillus flavus Plant disease-causing agent 2 Botrytis cinerea Gray mold 3 Cladosporium cladosporioides Flower mold 4 Rhizoctonia solani Leaf sheath blight fungus

[0124]

[0125] Experimental Example 1. Preparation of ethanol extracts from cassia seeds and cassia sprouts

[0126] Cassia tora sprouts include seeds soaked in water before germination, sprouted seeds, and cotyledons, and cassia tora sprouts before true leaves emerge were used. 10 times the weight of the dried crushed cassia tora sprouts and seeds was added with 100% ethanol (EtOH), and cold-extracted for 24 hours at 23-25℃ and 50-60% humidity. After filtration, the resulting mixture was concentrated under reduced pressure using a rotary evaporator, freeze-dried, and used in the experiment.

[0127]

[0128] Experimental Example 2. Preparation of solvent-specific fractions of ethanol extracts of Cassia tora seeds and Cassia tora sprouts

[0129] In Experimental Example 1, the freeze-dried product of the ethanol extracts of Cassia tora seeds and Cassia tora sprouts was dissolved or suspended by adding DW equivalent to 10 times the weight ratio. Afterwards, the mixture was placed in a separatory funnel and sequentially fractionated in the order of increasing non-polarity according to the order of polarity to obtain a total of five solvent fractions in the order of n-hexane, dichloromethane (MC), ethyl acetate (EtOAc), n-butanol (BuOH), and water (water, H2O).

[0130]

[0131] Experimental Example 3. Evaluation of antifungal activity of ethanol extracts of Cassia tora seeds and Cassia tora sprouts (MIC test)

[0132] Trichophyton rubrum was cultured on PDA (Potato dextrose Agar) medium (30℃, 1 week) to prepare fungi causing athlete's foot. The fungi cultured on the PDA plate in advance were cut into pieces with the agar and placed in SD (Sabouraud Dextrose) (liquid medium, conical tube), vortexed, and allowed to settle for 5 minutes. The supernatant (including spores) was diluted by concentration to prepare strain dilutions. In addition, the ethanol extracts of Cassia tora sprouts and Cassia tora seeds prepared in Experimental Example 1 were also diluted by concentration using SD medium (1% DMSO) and used.

[0133] In each well of a 96-well plate, 50 μL of the prepared strain dilution and 50 μL of the cassia tora sprout ethanol extract dilution or cassia tora seed ethanol extract dilution were mixed, the plate was covered, and the plate was incubated at 30°C for 1 week. The test concentration in the well where the growth of the fungus was inhibited was determined as the minimum inhibitory concentration (MIC).

[0134] Table 2 shows the results of confirming the minimum inhibitory concentration (MIC) against Trichophyton rubrum strains 4 days after treatment with ethanol extracts of Cassia tora sprouts and ethanol extracts of Cassia tora seed. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0135] As a result, the MIC value of the ethanol extract of Cassia tora sprouts was 500 ppm, which was one-fourth that of the ethanol extract of Cassia tora seed (2,000 ppm). Therefore, it can be seen that Cassia tora sprouts have antifungal activity four times better than Cassia tora seed.

[0136] In addition, the MIC value of the ethanol extract of Cassia tora seedlings decreased from 500 ppm to 62.5 ppm when treated with LED, confirming that the antifungal effect against Trichophyton rubrum strains was amplified 8-fold when treated with LED. On the other hand, the ethanol extract of Cassia tora seed did not show any amplification of the antifungal effect against Trichophyton rubrum strains according to LED treatment.

[0137] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueT.rubrumcrude extractSprout125-400050031.2-100062.5Seed2000-*MIC value: 4 days, more than 90% inhibition observed with the naked eye, *-: No inhibition observed

[0138]

[0139] Experimental Example 4. Evaluation of antifungal activity of solvent-specific fractions of ethanol extracts of Cassia tora seeds and Cassia tora sprouts against athlete's foot fungus (MIC test)

[0140] Fungi causing athlete's foot, such as E. floccosum, T. interdigitale, T. mentagrophytes, T. rubrum, T. verrucosum, or T. violaceum, were cultured (30°C, 1 week) on PDA (Potato dextrose Agar) medium to prepare the fungi. Diluted strains for E. floccosum, T. interdigitale, T. mentagrophytes, T. rubrum, T. verrucosum, or T. violaceum were prepared using the same method as Experimental Example 3.

[0141] The solvent fractions of the ethanol extract of Cassia tora sprouts and the ethanol extract of Cassia tora seed prepared in Experimental Example 2 were diluted to concentrations using SD medium (1% DMSO) and used as n-hexane, dichloromethane (MC), ethyl acetate (EtOAc), n-butanol (BuOH), and water (H2O) fractions.

[0142] In each well of a 96-well plate, 50 μL of the prepared strain dilution and 50 μL of the dilution of the fraction of the ethanol extract of Cassia tora sprouts or the dilution of the fraction of the ethanol extract of Cassia tora seed were mixed, and the plate was covered and incubated at 30°C for 1 week. The test concentration in the well where the growth of the athlete's foot fungus was inhibited was determined as the minimum inhibitory concentration (MIC).

[0143]

[0144] 4-1. Antifungal activity against E. floccosum

[0145] Table 3 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against E. floccosum strains after 5 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0146] As a result, it was confirmed that the hexane fraction (Hexane fr.) and dichloromethane fraction (MC fr.) of the ethanol extract of Cassia tora sprouts had superior antifungal effects against E. floccosum strains compared to the fractions of the ethanol extract of Cassia tora seed.

[0147] In addition, except for the water fraction of the ethanol extract of Cassia tora sprouts, the antifungal effect of the remaining solvent fractions against E. floccosum strains was amplified compared to before LED treatment, and it was confirmed that the antifungal effect of the solvent fractions of the ethanol extract of Cassia tora sprouts against E. floccosum strains was superior to that of the solvent fractions of the ethanol extract of Cassia tora seeds even when treated with LED.

[0148] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueE.floccosumHexane fr.sprout62.5-2000200015.6-50062.5Seed-125MC fr.sprout3.9-12562.51.9-62.515.6Seed12531.2EtOAc fr.sprout3.9-1251253.9-12562.5Seed125125BuOH fr.sprout156.2-5000500078.1-250062.5Seed5000125H20 fr.sprout156.2-5000-156.2-5000-Seed--*MIC value: 5 days, visual observation Inhibition of 90% or more, *-: No inhibition effect observed

[0149]

[0150] 4-2. Antifungal activity against T. interdigitale

[0151] Table 4 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. interdigitale strain after 6 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0152] As a result, it was confirmed that the hexane fraction (Hexane fr.) and dichloromethane fraction (MC fr.) of the ethanol extract of Cassia tora seed had a superior antifungal effect against the T. interdigitale strain compared to the fractions of the ethanol extract of Cassia tora seed. In addition, when treated with LED, the antifungal effect of all solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. interdigitale strain was amplified compared to before LED treatment, and it was confirmed that the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. interdigitale strain was superior to the solvent-specific fractions of the ethanol extract of Cassia tora seed even when treated with LED.

[0153] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueT.interdigitaleHexane fr.sprout62.5-200020007.8-25062.5Seed-250MC fr.sprout15.6-50062.50.9-31.2515.6Seed500-EtOAc fr.sprout7.8-250-0.4-15.62515.6Seed--BuOH fr.sprout156.2-5000-39.0-12501250Seed--H2O fr.sprout156.2-5000-156.2-50002500Seed--*MIC value: 6 days, more than 90% inhibition upon visual observation, *-: No inhibitory effect observed

[0154]

[0155] 4-3. Antifungal activity against T. mentagrophytes

[0156] Table 5 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. mentagrophytes strain after 5 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0157] As a result, it was confirmed that the hexane fraction (Hexane fr.), dichloromethane fraction (MC fr.), ethyl acetate fraction (EtOAc fr.), and butanol fraction (BuOH fr.) of the ethanol extract of Cassia tora sprouts had a superior antifungal effect against T. mentagrophytes strains compared to the fractions of the ethanol extract of Cassia tora seed.

[0158] In addition, except for the water fraction of the ethanol extract of Cassia tora seed, the antifungal effect of the remaining solvent fractions on T. mentagrophytes strains was amplified compared to before LED treatment, and it was confirmed that the antifungal effect of the solvent fractions of the ethanol extract of Cassia tora seed on T. mentagrophytes strains was superior to that of the solvent fractions of the ethanol extract of Cassia tora seed even when treated with LED.

[0159] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueT. mentagrophytesHexane fr.sprout125-4000100031.2-100062.5Seed--MC fr.sprout15.6-5001250.9-31.257.8Seed500-EtOAc fr.sprout15.6-50062.50.9-31.2515.6Seed250-BuOH fr.sprout312.5-100001000039.0-125062.5Seed--H2O fr.sprout312.5-10000-31.2-10000-Seed--*MIC value: 5 days, more than 90% upon visual observation Inhibition, *-: No inhibitory effect observed

[0160]

[0161] 4-4. Antifungal activity against T. rubrum

[0162] Table 6 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. rubrum strain after 5 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0163] As a result, it was confirmed that the hexane fraction (Hexane fr.), dichloromethane fraction (MC fr.), and ethyl acetate fraction (EtOAc fr.) of the ethanol extract of Cassia tora sprouts had a superior antifungal effect against T. rubrum strains compared to the fractions of the ethanol extract of Cassia tora seed.

[0164] In addition, the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts on T. rubrum strains was amplified compared to before LED treatment, and it was confirmed that the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts on T. rubrum strains was superior to that of the solvent-specific fractions of the ethanol extract of Cassia tora seeds even when treated with LED.

[0165] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueT.rubrumHexane fr.sprout62.5-2000200031.2-1000250Seed-1000MC fr.sprout78.1-250062.53.9-12515.625Seed250125EtOAc fr.sprout3.9-1251253.9-12515.625Seed--BuOH fr.sprout156.2-500078.1-25001250Seed--H2O fr.sprout156.2-5000156.2-50002500Seed--*MIC value: 5 days, more than 90% upon visual observation Inhibition, *-: No inhibitory effect observed

[0166]

[0167] 4-5. Antifungal activity against T. verrucosum

[0168] Table 7 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. verrucosum strain after 4 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0169] As a result, it was confirmed that the hexane fraction (Hexane fr.), dichloromethane fraction (MC fr.), and ethyl acetate fraction (EtOAc fr.) of the ethanol extract of Cassia tora sprouts had superior antifungal effects against the T. verrucosum strain compared to the fractions of the ethanol extract of Cassia tora seed.

[0170] In addition, the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts on T. verrucosum strains was amplified compared to before LED treatment, and it was confirmed that the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts on T. verrucosum strains was superior to that of the solvent-specific fractions of the ethanol extract of Cassia tora seeds even when treated with LED.

[0171] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueT. verrucosumHexane fr.sprout31.2-1000100031.2-100062.5Seed--MC fr.sprout7.8-25062.57.8-25015.6Seed25062.5EtOAc fr.sprout7.8-25062.57.8-25015.6Seed25062.5BuOH fr.sprout156.2-500-78.1-2500312.5Seed--H20 fr.sprout156.2-500-156.2-5001250Seed--*MIC value: 4th day, more than 90% inhibition upon visual observation, *-: No inhibitory effect observed

[0172]

[0173] 4-6. Antifungal activity against T.violaceum

[0174] Table 8 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the T. violaceum strain after 8 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0175] As a result, it was confirmed that the hexane fraction (Hexane fr.), dichloromethane fraction (MC fr.), ethyl acetate fraction (EtOAc fr.), and butanol fraction (BuOH fr.) of the ethanol extract of Cassia tora sprouts had a superior antifungal effect against the T. violaceum strain compared to the fractions of the ethanol extract of Cassia tora seed.

[0176] In addition, the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts on T. violaceum strains was amplified compared to before LED treatment, and it was confirmed that the antifungal effect of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts on T. violaceum strains was superior to that of the solvent-specific fractions of the ethanol extract of Cassia tora seeds even when treated with LED.

[0177] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueT.violaceumHexane fr.sprout125-4000100031.2-100031.2Seed-250MC fr.sprout15.6-50031.20.9-31.253.9Seed25015.6EtOAc fr.sprout15.6-5001250.9-31.257.8Seed25031.2BuOH fr.sprout31.2-10000250039.0-125031.2Seed10000-H20 fr.sprout31.2-10000-31.2-1000031.2Seed-1250*MIC value: 8 Primary, more than 90% inhibition upon visual observation, *-: No inhibition effect observed

[0178]

[0179] Experimental Example 5. Evaluation of antifungal activity of solvent-specific fractions of ethanol extracts of Cassia tora seeds and Cassia tora sprouts against human disease-causing bacteria (MIC test)

[0180] A. fumigatus or C. albicans was cultured (30°C, 1 week) on PDA (Potato dextrose Agar) medium to prepare the strain dilution solution for A. fumigatus or C. albicans using the same method as Experimental Example 3.

[0181] The solvent fractions of the Cassia tora sprout ethanol extract and Cassia tora seed ethanol extract prepared in Experimental Example 2, n-hexane, dichloromethane (MC), ethyl acetate (EtOAc), n-butanol (BuOH), and water (H2O) fractions were diluted to different concentrations using SD medium (1% DMSO) and used.

[0182] In each well of a 96-well plate, 50 μL of the prepared strain dilution and 50 μL of the dilution of the fraction of the ethanol extract of Cassia tora sprouts or the dilution of the fraction of the ethanol extract of Cassia tora seed were mixed, and the plate was covered and incubated at 30°C for 1 week. The test concentration in the well where the growth of the athlete's foot fungus was inhibited was determined as the minimum inhibitory concentration (MIC).

[0183]

[0184] 5-1. Antifungal activity against A. fumigatus

[0185] Table 9 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against A. fumigatus strains after 5 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0186] As a result, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against A. fumigatus strains. In addition, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against A. fumigatus strains even when treated with LED.

[0187] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueA. fumigatusHexane fr.sprout62.5-2000-15.6-500-seed--MC fr.sprout15.6-500-3.9-125-seed--EtOAc fr.sprout15.6-500-3.9-125-seed--BuOH fr.sprout78.1-2500-78.1-2500-seed--H2O fr.sprout78.1-2500-78.1-2500-seed--*MIC value: 4 days, more than 90% inhibition observed with the naked eye, *-: No inhibition observed

[0188]

[0189] 5-2. Antifungal activity against C. albicans

[0190] Table 10 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the C. albicans strain after 3 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0191] As a result, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against C. albicans strains. In addition, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against C. albicans strains even when treated with LED.

[0192] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueC. albicansHexane fr.sprout62.5-2000-15.6-500-seed--MC fr.sprout15.6-500-3.9-125-seed--EtOAc fr.sprout15.6-500-3.9-125-seed--BuOH fr.sprout78.1-2500-78.1-2500-seed--H2O fr.sprout78.1-2500-78.1-2500-seed--*MIC value: 3 days, more than 90% inhibition observed with the naked eye, *-: No inhibition observed

[0193]

[0194] Experimental Example 6. Evaluation of antifungal activity of solvent-specific fractions of ethanol extracts of Cassia tora seeds and Cassia tora sprouts against plant disease-causing fungi (MIC test)

[0195] A. flavus, B. cinerea, C. cladosporiodes, or R. solani were cultured (30°C, 1 week) on PDA (Potato dextrose Agar) medium to prepare the strain dilution solution for A. flavus, B. cinerea, C. cladosporiodes, or R. solani using the same method as Experimental Example 3.

[0196] The solvent fractions of the Cassia tora sprout ethanol extract and Cassia tora seed ethanol extract prepared in Experimental Example 2, n-hexane, dichloromethane (MC), ethyl acetate (EtOAc), n-butanol (BuOH), and water (H2O) fractions were diluted to different concentrations using SD medium (1% DMSO) and used.

[0197] In each well of a 96-well plate, 50 μL of the prepared strain dilution and 50 μL of the dilution of the fraction of the ethanol extract of Cassia tora sprouts or the dilution of the fraction of the ethanol extract of Cassia tora seed were mixed, and the plate was covered and incubated at 30°C for 1 week. The test concentration in the well where the growth of the athlete's foot fungus was inhibited was determined as the minimum inhibitory concentration (MIC).

[0198]

[0199] 6-1. Antifungal activity against A. flavus

[0200] Table 11 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against A. flavus strains after 3 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0201] As a result, it was confirmed that the solvent-specific fractions of the ethanol extract of Cassia tora and the solvent-specific fractions of the ethanol extract of Cassia tora had no antifungal effect against A. flavus strains. However, the dichloromethane fraction (MC fr.) of the ethanol extract of Cassia tora under LED treatment had an antifungal effect against A. flavus strains.

[0202] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueA.flavusHexane fr.sprout62.5-2000-15.6-500-seed--MC fr.sprout15.6-500-3.9-125125-seed--EtOAc fr.sprout15.6-500-3.9-125-seed--BuOH fr.sprout78.1-2500-78.1-2500-seed--H2O fr.sprout78.1-2500-78.1-2500-seed--*MIC value: 3 days, more than 90% inhibition observed with the naked eye, *-: No inhibition observed

[0203]

[0204] 6-2. Antifungal activity against B. cinerea

[0205] Table 12 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against B. cinerea strains after 3 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0206] As a result, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against B. cinerea strains. In addition, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against B. cinerea strains even when treated with LED.

[0207] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueB.cinereaHexane fr.sprout62.5-2000-15.6-500-seed--MC fr.sprout15.6-500-3.9-125-seed--EtOAc fr.sprout15.6-500-3.9-125-seed--BuOH fr.sprout78.1-2500-78.1-2500-seed--H20 fr.sprout78.1-2500-78.1-2500-seed--*MIC value: 3 days, more than 90% inhibition observed with the naked eye, *-: No inhibition observed

[0208]

[0209] 6-3. Antifungal activity against C. cladosporiodes

[0210] Table 13 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the C. cladosporiodes strain after 2 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0211] As a result, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against C. cladosporiodes strains. In addition, it was confirmed that the solvent fractions of the ethanol extract of Cassia tora sprouts and the solvent fractions of the ethanol extract of Cassia tora seed had no antifungal effect against B. cinerea strains even when treated with LED.

[0212] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueC. cladosporiodesHexane fr.sprout62.5-2000-15.6-500-seed--MC fr.sprout15.6-500-3.9-125-seed--EtOAc fr.sprout15.6-500-3.9-125-seed--BuOH fr.sprout78.1-2500-78.1-2500-seed--H20 fr.sprout78.1-2500-78.1-2500-seed--*MIC value: 2 days, more than 90% inhibition observed with the naked eye, *-: No inhibition observed

[0213]

[0214] 6-4. Antifungal activity against R. solani

[0215] Table 14 shows the results of examining the minimum inhibitory concentration (MIC) of the solvent-specific fractions of the ethanol extract of Cassia tora sprouts and the solvent-specific fractions of the ethanol extract of Cassia tora seed against the R. solani strain after 5 days. The LED treatment group was treated with LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for 2 days.

[0216] As a result, it was confirmed that the hexane fraction (Hexane fr.), dichloromethane fraction (MC fr.), and ethyl acetate fraction (EtOAc fr.) of the ethanol extract of Cassia tora sprouts had a superior antifungal effect against the R. solani strain compared to the fractions of the ethanol extract of Cassia tora seed.

[0217] In addition, except for the water fraction, the solvent fractions of the ethanol extract of Cassia tora seed showed an amplified antifungal effect against the R. solani strain compared to before the LED treatment, and it was confirmed that the antifungal effect of the solvent fractions of the ethanol extract of Cassia tora seed against the R. solani strain was superior to that of the solvent fractions of the ethanol extract of Cassia tora seed even after the LED treatment.

[0218] OrganismAntifungalsBroth microdilution (ppm)LED untreated groupLED treated groupRangeMIC valueRangeMIC valueRangeMIC valueR.solaniHexane fr.sprout62.5-2000100062.5-2000125Seed-250MC fr.sprout15.6-50012515.6-50031.2Seed500125EtOAc fr.sprout7.8-2501257.8-25015.6Seed-250BuOH fr.sprout78.1-2500-78.1-2500312.5Seed--H20 fr.sprout78.1-2500-78.1-2500-Seed--*MIC value: 5 days, more than 90% inhibition observed with the naked eye, *-: Inhibitory effect observed No

[0219]

[0220] Experimental Example 7. Chromatogram analysis of fractions from ethanol extracts of Cassia tora sprouts

[0221] As shown in Tables 3 to 8 above, it was confirmed that the dichloromethane fraction (MC fr.) of the ethanol extract of Cassia tora sprouts exhibited excellent antifungal activity at the lowest concentration in the LED-treated group as well as the LED-untreated group against athlete's foot fungi such as E. floccosum, T. interdigitale, T. mentagrophytes, T. rubrum, T. verrucosum, or T. violaceum.

[0222] Therefore, HPLC analysis was performed to isolate compounds exhibiting antifungal activity from the dichloromethane fraction (MC fr.) of the ethanol extract of Cassia tora sprouts. Medium pressure liquid chromatography (MPLC) was performed to trace substances with antifungal activity from the major peaks identified by HPLC (High Performance Liquid Chromatography). Subsequently, HPLC-DAD, LC-MS, and NMR analyses were performed to identify seven compounds, as shown in Table 15 and Figure 1. Figure 1 shows the HPLC-DAD (High-performance liquid chromatography with diode-array detection) of the dichloromethane fraction (MC fr.) of Cassia tora sprouts.

[0223] Tentative identification[LC-DAD]RT(min)Molecularweight(m / z) UV (nm)[M+H]+[MH]-Aurantio-obtusin14.8330331329.1283, 393Macrosporin16.2284-283.1223, 276, 411Chrysoobtusin16.6358359.2357.1282, 359Obtusin17.8344345343283, 316, 385Emodin19.7270-269.1219, 286, 437Rubrofusarin21.3272-303223, 275, 324, 404Chrysophanol23.6254-253223, 255, 286, 427

[0224]

[0225] Experimental Example 8. Analysis of antifungal activity of compounds isolated from fractions of ethanol extract of Cassia tora sprouts.

[0226] The antifungal activity of Aurantio-obtusin, Macrosporin, Chrysoobtusin, Obtusin, Emodin, Rubrofusarin, and Chrysophanol, which were separated by chromatographic fractionation using the dichloromethane fraction (MC fr.) of the ethanol extract of Cassia tora sprouts, was evaluated in the same manner as in Experimental Example 4.

[0227] Table 16 shows the results of examining the minimum inhibitory concentration (MIC) of seven compounds isolated from the dichloromethane fraction (MC fr.) against T. rubrum. The LED treatment group was exposed to LED light (455±2 nm) for 15 minutes per day (15 minutes / day) for two days.

[0228] As a result, it can be seen that Aurantio-obtusin and Obtusin compounds have antifungal activity, but the MIC values ​​are the same for both the LED treatment and untreated groups, indicating that they do not react to LED. However, the Emodin compound has an MIC value of 250 ppm when not treated with LED, but when treated with LED, the minimum inhibitory concentration (MIC) decreases by 16-fold to 15.6 ppm, indicating that it corresponds to a compound whose antifungal activity is enhanced when treated with LED. Chrysophanol has no antifungal inhibition activity when not treated with LED, but shows strong antifungal activity at 7.8 ppm when treated with LED, indicating that it corresponds to a compound that shows antifungal activity by LED treatment.

[0229] MPLC Separation Compound Broth microdilution (ppm) Note (Antifungal activity) Range LED Untreated group MIC value LED Treated group MIC value Aurantio-obtusin 15.6-1000 31.2 31.2 Schedule Macrosporin---chrysoobtusin-250 4-fold or more increase Obtusin 1000 1000 Schedule Emodin 250 15.6 16-fold increase Rubrofusarin---Chrysophanol-7.8 128-fold or more increase

[0230] Next, to verify whether the compounds isolated from Cassia tora sprouts and actual compounds exhibit the same antifungal activity, an antifungal experiment was conducted on commercially available compounds. As a result, the Aurantio-obtusin compound showed no change in antifungal activity between LED treatment and untreated conditions, consistent with the results confirmed in Table 16. The Emodin compound had an MIC value of 250 ppm when untreated with LED, but when treated with LED, the MIC value decreased by 16-fold to 15.6 ppm. In addition, chrysophanol had no antifungal inhibition activity when untreated with LED, but when treated with LED, the MIC value was 7.8 ppm, showing the same antifungal activity.

[0231] Purchased compound (commercially available compound) Broth microdilution (ppm) Note (antifungal activity) Range LED Untreated MIC value LED Treated MIC value Aurantio-obtusin 0.9-500 125 125 Schedule Emodin 250 15.6 16-fold increase Chrysophanol - 7.8 128-fold increase

[0232]

[0233] 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 made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

Claims

1. An antifungal composition comprising an extract of cassia seed sprouts or a fraction thereof as an active ingredient.

2. An antifungal composition, wherein the cassia seed sprout extract or a fraction thereof in paragraph 1 exhibits photosensitivity under light conditions having a wavelength of 400 nm to 528 nm.

3. In paragraph 1, The above cassia seed sprout extract is an antifungal composition obtained by extracting cassia seed sprouts with water, C1 to C4 alcohol, or a mixed solvent thereof.

4. In paragraph 1, An antifungal composition, wherein the above cassia seedlings include seeds before germination, germinated seeds, and germinated cotyledons after being soaked in water.

5. In paragraph 1, The above fraction is an antifungal composition prepared by fractionating the above cassia seed sprout extract with water, an alcohol having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate, butanol or a mixed solvent thereof.

6. In paragraph 1, An antifungal composition, wherein the compound separated from the above fraction is at least one compound selected from the group consisting of aurantio-optusin, emodin, and chrysophanol.

7. In paragraph 2, The luminous intensity of the above light is 10 to 2,000 μmol m -2 s -1 An antifungal composition, which is investigated as.

8. In paragraph 2, An antifungal composition wherein the light is irradiated at a distance of 1 to 30 cm.

9. In paragraph 2, An antifungal composition, wherein the light is irradiated 1 to 6 times a day for 5 to 20 minutes.

10. In paragraph 1, An antifungal composition having antifungal activity against Trichophyton, eflatoxin-producing bacteria or leaf sheath blight bacteria.

11. In paragraph 10, An antifungal composition, wherein the above-mentioned trichophyton is selected from the group consisting of Epidermophyton floccosum, Trichophyton interdigitale, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton verrucosum, and Trichophyton violaceum.

12. In paragraph 10, An antifungal composition, wherein the above eflatoxin-producing bacteria is Aspergillus flavus.

13. In paragraph 10, An antifungal composition, wherein the above leaf sheath blight fungus is Rhizoctonia solani.

14. In any one of paragraphs 1 to 11, The antifungal composition is applied as one or more selected from the group consisting of a pharmaceutical composition for treating or preventing tinea, a food composition for improving or preventing tinea, a food additive composition for improving or preventing tinea, a quasi-drug composition for improving or preventing tinea, a cosmetic composition for improving or preventing tinea, and a skin external application composition for treating or preventing tinea.

15. A step of applying a composition of any one of claims 1 to 11 to the skin or nails of an individual; and A method for preventing or treating tinea, comprising the step of irradiating the skin or fingernails of an individual with light having a wavelength of 400 nm to 528 nm.

16. Step 1 of manufacturing cassia seed sprout extract; Step 2: fractionating the above Cassia tora sprout extract with water, alcohol having 1 to 4 carbon atoms, hexane, dichloromethane, ethyl acetate or a mixed solvent thereof to prepare a fraction of the Cassia tora sprout extract; and A method for isolating a compound having antifungal activity, comprising the step of isolating at least one compound selected from the group consisting of aurantio-optusin, emodin, and chrysophanol from a fraction of the above cassia seedling extract.

Images