KR20190135448A - A composition having anti-bacterial activity comprising Selaginella tamariscina extracts, fractions thereof or compounds isolated therefrom as an active ingredient - Google Patents

Abstract

The present invention relates to an antioxidant or antibacterial cosmetic composition, a food composition, and a pharmaceutical composition, wherein the antioxidant or antibacterial cosmetic composition comprises a Selaginella tamariscina extract, a fraction thereof, or a compound isolated therefrom, as an active ingredient. The antioxidant or antibacterial cosmetic composition comprising a Selaginella tamariscina extract, a fraction thereof, or a compound isolated therefrom, as an active ingredient has an excellent concentration-dependent DPPH free radical scavenging activity, that is, antioxidant effect, and has an excellent inhibitory activity on the growth of bacteria causing food poisoning, dermatitis, pneumonia, cellulitis, etc., and thus can be beneficially used in cosmetic products, food, drugs, etc. for preventing, alleviating, or treating a disease caused by oxidation and bacterial infections.

Description

A composition having anti-bacterial activity comprising Selaginella tamariscina extracts, fractions particular or compounds isolated therefrom as an active ingredient

The present invention relates to an antioxidant or antimicrobial cosmetic composition, food composition and pharmaceutical composition comprising Selaginella tamariscina extract, fractions thereof or compounds isolated therefrom as an active ingredient.

Oxygen is essential for various metabolic reactions to maintain life, and is also necessary for the detoxification of poisonous substances in the human body, but oxygen is not only beneficial to the human body, so various pollutants such as enzymes, reducing metabolism, chemicals, photochemical reactions, and physical chemicals When converted to highly reactive free radicals such as superoxide radicals, hydroxyl radicals, hydrogen peroxide and singlet oxygen by environmental factors, Bilaterally causing fatal oxygen toxicity in living organisms ( Korea J. Biotechnol. Bioeng. , 2001, 16 (6), 592-602). Synthetic antioxidants such as butylated hydroxy toluene (BHT) and butylated hydroxy anisole (BHA), which have been used widely to remove these active oxygen, have been widely used until now because of their excellent antioxidant effects and economic efficiency. This raises the issue of limiting the amount of food or use allowed. In addition, natural antioxidants, such as tocopherol and ascorbic acid, have high safety, but alone have a low chain arrest ability and high cost. For this reason, recent studies have been actively conducted to separate and use antioxidants that are safer, more economical, and more effective from natural products. In particular, plant-derived secondary metabolites can inhibit the production of free radicals and active oxygen Much research has been done on this because it prevents oxidation by removing it ( Korean Journal of Plant Res. , 2011, 24 (1), 30-39; J. Korean Soc. Food Sci. Nutr. , 2010, 39 (9) ), 1249-1256).

On the other hand, food, cosmetics, textiles and pharmaceutical products are necessary to maintain the quality for a long time, preservatives to prevent decay by microorganisms is essential, especially in the case of cosmetic products with a long shelf life and a lot of microbial nutrients in the nature of the product more preservatives Is required. As recently raised in the Oxy Case, there are always risks compared to the economics of synthetic raw materials, and the problem that arises when risks are realized is fatal to assess profits based on simple arithmetic statistics.

Parabens preservatives most commonly used in cosmetics, foods and medicines as existing preservatives have been proposed to cause skin allergies and resistant bacteria, and researches on natural preservatives with excellent safety and economical efficiency to solve these problems. Is continuing. Natural preservatives (preservatives) are additives used for long-term preservation and shelf life in foods and cosmetics, and are bio-derived materials used to prevent decay by microorganisms. Despite the strict limitations, concerns about various side effects, such as chronic and acute toxicity, mutations and carcinogens, are constantly being raised.

Most of the currently reported natural antimicrobial substances are not commercialized due to color odor, deterioration in stability, narrow antibacterial spectrum and formulation problems, such as hinokitiol which is a protein extract, magnolol which is a magnolia extract, and grapefruit. Only a few, such as seed extract DF-100, have been commercialized. In the case of DF-100, which is the most advanced in product development, the antimicrobial activity is known to be due to benzethonium chloride, which is an organic acid and synthetic preservative contained in DF-100, and other natural antibacterial agents have low economical efficiency. In addition, there is a problem such as limited antimicrobial spectrum or limited use range due to physical properties, which is an urgent need for natural antimicrobial agents that can be more developed and commercialized (Korea Patent No. 10-1495272).

Under these circumstances, the present inventors have made efforts to find natural products with excellent antioxidant or antimicrobial activity without side effects on the human body, and have been prescribed for various bleeding symptoms such as hemorrhage, stool bleeding, and uterine bleeding, and to relieve pain caused by dysmenorrhea or bruises The saturated butanol fraction obtained from the methanol extract of Selaginella tamariscina known to be used has excellent DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging effect, dermatitis, The present invention was completed by confirming that the effect of inhibiting microbial growth associated with disease induction such as food poisoning and pneumonia is excellent.

Korean Patent Registration No. 10-1495272

Korea J. Biotechnol. Bioeng., 2001, 16 (6), 592-602 Korean Journal of Plant Res., 2011, 24 (1), 30-39 J. Korean Soc. Food Sci. Nutr., 2010, 39 (9), 1249-1256

An object of the present invention is to provide an antioxidant or antimicrobial composition containing the saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient.

Another object of the present invention is to provide a cosmetic composition comprising the composition for antioxidant or antibacterial.

Another object of the present invention is to provide a food composition comprising the composition for antioxidant or antibacterial.

Another object of the present invention is to provide a pharmaceutical composition comprising the composition for antioxidant or antibacterial.

In order to achieve the above object, the present invention is an antioxidant or antimicrobial composition containing a saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient, a cosmetic composition, a food composition and a pharmaceutical composition comprising the antioxidant or antimicrobial composition To provide a composition.

Hereinafter, the present invention will be described in detail.

In one embodiment, the present invention provides an antioxidant or antimicrobial composition containing a saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient.

The term " Selaginella tamariscina " of the present invention is a fern that grows on mountain-grown rocks and is sometimes called Kwonbaek. A medicinal plant belonging to the Selaginellaceae family, with a unique appearance resembling the hand of the Buddha. The scientific name is Selaginella tamariscina (P.Beauv.) Spring.

Specifically, the leaves are dense in four rows, ovate, 1.5 ~ 2 mm long, and end of a thread-like protrusion, and there is a cup holder at the most geography. Spores are egg-shaped triangles, with fine and thin spines, with spores large and small. Spore sac (胞子 囊 穗) hangs at the end of twigs one by one, square, 5-15mm long, 2mm in diameter. The stem reaches 20cm in height, the branch is divided into planes, the surface is dark green, and the back side is white green. Branches spread in all directions when wet, and curled into balls when dry, and spread again when there is moisture. Roots grow with many dull bodies and roots tangled together, with branches spreading out from the ends formed like stems. Root-derived roots are hard and short and have many beard roots.

It is almost evident in Korea, distributed in Japan, Taiwan, China, Manchuria, etc. It is an evergreen perennial plant that grows mainly on dry and cool rocks or cliff surfaces. It has been used to treat bloody stools, hematuria, and anal hernias in oriental medicine. As the blood sugar drop is known, interest in the efficacy of kwonbaek is increasing.

The saturated butanol fraction of the methanol extract of Selaginella tamariscina is preferably prepared by a manufacturing method comprising the following steps:

1) extracting by adding methanol to Selaginella tamariscina ;

2) filtering the extract of step 1);

3) concentrating the filtered extract of step 2) under reduced pressure and drying to prepare a Buddha extract methanol; And

4) preparing the butcherson fraction by further extracting the butcherson methanol extract of step 3) with an organic solvent.

In the method, Selaginella tamariscina of step 1) can be used without limitation, such as grown or commercially available. In addition, in step 1), but not limited to this, the Buddha hand may be to extract all, including the leaves, branches or roots.

In the above method, as the extraction method using methanol of step 1), it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but is not limited thereto. The methanol is preferably extracted by adding 1 to 10 times the amount of the washed, dried and powdered Buddha. The extraction temperature is preferably 20 ° C to 100 ° C, more preferably 20 ° C to 60 ° C, and most preferably 50 ° C, but is not limited thereto. In addition, the extraction time is preferably 4 to 24 hours, more preferably 5 to 12 hours, most preferably 6 hours, but is not limited thereto. In addition, the number of extraction is preferably 1 to 5 times, more preferably repeated extraction 2 to 4 times, most preferably 3 times, but is not limited thereto.

In the above method, the decompression concentration in step 3) preferably uses a vacuum decompression concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

In the method, the organic solvent of step 4) is n-hexane preferably in the (n -Hexane), methylene chloride (methylene chloride), ethyl acetate (ethyl acetate), or saturated n- butanol (water saturated n-butanol) However, this is not limitative. The fraction was then suspended in water bucheoson methanol extract of n-hexane (n -Hexane), methylene chloride (methylene chloride), ethyl acetate (ethyl acetate), saturated n- butanol (water saturated n-butanol) and water ( It is preferable that the normal-hexane fraction, methylene chloride fraction, ethyl acetate fraction, water saturated n-butanol fraction or water fraction obtained by systematic fractionation with H ₂ O), and water fraction, More preferably, it is a saturated n-butanol fraction, but is not limited thereto. The fraction may be obtained by repeating the fractionation process from 1 to 5 times, preferably 3 times, from the Busonson methanol extract, and preferably concentrated under reduced pressure after the fraction, but is not limited thereto.

In addition, the saturated butanol fraction of the methanol extract of Selaginella tamariscina is preferably prepared by a manufacturing method including the following steps:

1) extracting by adding methanol to Selaginella tamariscina ;

2) filtering the extract of step 1);

3) concentrating the filtered extract of step 2) under reduced pressure and drying to prepare a Buddha extract methanol; And

4) step of extracting the butcherson methanol extract of step 3) with additional saturated butanol to prepare the butcherson saturated butanol fraction.

In the method, Selaginella tamariscina of step 1) can be used without limitation, such as grown or commercially available. In addition, in step 1), but not limited to this, the Buddha hand may be to extract all, including the leaves, branches or roots.

In the above method, as the extraction method using methanol of step 1), it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but is not limited thereto. The methanol is preferably extracted by adding 1 to 10 times the amount of the washed, dried and powdered Buddha. The extraction temperature is preferably 20 ° C to 100 ° C, more preferably 20 ° C to 60 ° C, and most preferably 50 ° C, but is not limited thereto. In addition, the extraction time is preferably 4 to 24 hours, more preferably 5 to 12 hours, most preferably 6 hours, but is not limited thereto. In addition, the number of extraction is preferably 1 to 5 times, more preferably repeated extraction 2 to 4 times, most preferably 3 times, but is not limited thereto.

In the above method, the decompression concentration in step 3) preferably uses a vacuum decompression concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

In the above method, the butadiene-saturated butanol fraction of step 4) is preferably fractionated by adding the same amount of saturated saturated n-butanol to the butyric methanol extract. The fraction may be obtained by repeating the fractionation process from 1 to 5 times, preferably 3 times, from the Busonson methanol extract, and preferably concentrated under reduced pressure after the fraction, but is not limited thereto.

In addition, although not limited thereto, the saturated n-butanol is mixed with butanol and ultrapure water in the same amount, violently shaken and left to completely separate the layers, and then discards the lower water layer and recovers the upper layer. Be prepared.

On the other hand, but is not limited to the saturated butanol fractions α-linolenic acid (α-linolenic acid), 3-deoxy-d-mannoic acid (3-Deoxy-d-mannoic acid), guaiacol (guaiacol) And butanediol compounds.

In addition, the compound is not limited thereto, and the compound has 5 to 10 parts by weight of α-linolenic acid and 3-dioxy-di-mannoic acid based on 100 parts by weight of the saturated butanol fraction. 3 to 6 parts by weight of deoxy-d-mannoic acid, 5 to 10 parts by weight of guaiacol, and 0.1 to 3 parts by weight of butanediol.

The term "antioxidant" of the present invention refers to the action of inhibiting oxidation, the human body is a balance between the antioxidant (prooxidant) and antioxidant (antioxidant), but due to a number of factors this balance state is imbalanced When inclined towards oxidative stress, oxidative stress is induced, leading to potential cell damage and pathological disease. Reactive oxygen species (ROS), which is a direct cause of this oxidative stress, are unstable and highly reactive, easily react with various biological substances, attack polymers in the body, causing irreversible damage to cells and tissues, or mutations, It results in cytotoxicity and carcinogenesis. Reactive nitrogen species (RNS), such as NO, HNO ₂ and ONOO-, are produced in large amounts due to the immune response of macrophage neutrophils and other immune cells during inflammatory reactions. Such active oxygen oxidizes and destroys cells in the body, thereby exposing them to various diseases. Therefore, the inclusion of the Busonson methanol extract of the present invention, in particular the saturated butanol fraction of the Busonson methanol extract, such as cosmetics, food or pharmaceuticals to achieve an antioxidant effect, it can contribute to anti-aging and health promotion.

In a specific embodiment of the present invention, in order to confirm the antioxidant effect of the saturated butanol fraction of the Buddha extract methanol, DPPH free radical scavenging activity measurement and metal ion catalyst oxidation inhibition test, as a result of the saturated butanol fraction of Buddha extract methanol It was confirmed that the concentration-dependent scavenging effect and protein protection effect of DPPH free radicals, that is, exhibiting strong antioxidant effect (FIG. 5 and FIG. 6). Therefore, the antioxidant effect of the present invention may be achieved by free radical scavenging activity.

The term "antibacterial" of the present invention means the action of inhibiting or controlling the growth and proliferation of microorganisms such as bacteria or fungi, the microorganism of the present invention is not limited thereto, Staphylococcus aureus ( Staphylococcus aureus ), Star may be a pillow nose kusu epidermidis (Staphylococcus epidermidis), or Haemophilus influenzae (Haemophilus influenzae). The skin flora, Staphylococcus aureus , is generally present on the skin and nasal surfaces of healthy people or livestock, and is known as a bacterium that produces food poisoning by producing heat-resistant exotoxins. Staphylococcus epidermidis ( Staphylococcus epidermidis ) is a secondary skin infection after acne bacteria infection, which is generally non-pathogenic, sepsis, urinary tract infections, and endocarditis. Haemophilus influenzae, the causative agent of influenza , is known to cause bacterial meningitis or acute epiglottitis.

On the other hand, various types of skin diseases are mainly caused by dermatophytes, such as acne bacteria, dandruff bacteria. The onset of acne has been reported as a major factor causing the inflammatory response of acne bacteria, dandruff bacteria ideally proliferate in the skin such as the back, nape, causing seborrheic dermatitis. The cause of atopic dermatitis is not known yet, but it is reported as staphylococci as a causative agent of onset. The use of preservatives and antimicrobial agents is essential to reduce the occurrence of skin diseases caused by these dermatophytes and other bacteria and to protect the skin, but the synthetic materials used in the past may cause allergies to the skin, so they are relatively harmless to humans. It is important to do.

Accordingly, the inclusion of the Busonson methanol extract of the present invention, in particular the saturated butanol fraction of the methanol extract in cosmetics, food or pharmaceuticals, including microorganisms that induce dermatitis, including food poisoning and pneumonia By achieving the effect of inhibiting microbial growth, it can contribute to the prevention and improvement of diseases such as food poisoning, pneumonia, including skin diseases.

In a specific embodiment of the present invention, in order to confirm the antimicrobial effect of the saturated butanol fraction of the Buddha extract methanol extract, dermatitis, food poisoning, pneumonia by using a minimum inhibitory concentration assay ( Staphylococcus aureus associated with the induction of pneumonia, Staphylococcus epidermidis (Staphylococcus epidermidis) and Haemophilus influenzae (Haemophilus influenzae) check result of measuring the antimicrobial activity for the three strains in total, which bucheoson saturated butanol fraction the growth inhibitory activity is superior with respect to the three strains 10 and 11. Therefore, the antimicrobial effect of the present invention may be achieved by growth inhibitory activity against food poisoning bacteria and influenza bacteria, including skin microorganisms.

As another aspect, the present invention provides a cosmetic composition comprising an antioxidant or antimicrobial composition containing a saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient.

The saturated butanol fraction of the butanone methanol extract is as described above, and the saturated butanol fraction of the butanone methanol extract of the present invention can be added to the cosmetic composition for the purpose of antioxidant or antibacterial since it has an antioxidant effect and an antimicrobial effect. have.

The cosmetic composition prevents or improves any one skin disease selected from the group including acne, atopy, athlete's foot, psoriasis, eczema and dermatitis, and has the effect of preventing or improving skin aging, wrinkles or loss of elasticity.

In a specific embodiment of the present invention, the saturated butanol fraction of Buddha extract methanol has excellent antioxidant effect and Staphylococcus aureus , Staphylococcus epidermidis (Staphylococcus epidermidis) and Haemophilus influenzae (Haemophilus influenzae) was confirmed to show the antimicrobial effect of suppressing the growth of the strains (5, 6, 10 and 11).

The present invention provides an antimicrobial effect of the saturated butanol fraction of Busonson methanol extract with excellent growth inhibitory activity against food poisoning bacteria and influenza bacteria, including skin microorganisms. While antibiotics such as tetracycline have limitations in use, such as the side effects and the emergence of resistant bacteria, the saturated butanol fraction of the Buddha extract methanol according to the present invention is safe and has no side effects, and thus is suitable as a cosmetic composition.

The cosmetic composition of the present invention can be prepared in the form of general emulsion formulations and solubilized formulations. The emulsified formulations include nutrient cosmetics, creams, essences, etc., and the solubilized formulations include soft cosmetics. Suitable formulations include, but are not limited to, solutions, gels, solid or pasty anhydrous products, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionics (liposomes) obtained by dispersing an oil phase in an aqueous phase, for example In the form of a vesicle dispersant, may be in the form of a cream, skin, lotion, powder, ointment, spray or cone stick. It may also be in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

The cosmetic composition additionally contains fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, blowing agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metals Commonly used adjuvants such as ion blockers, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetic compositions It may contain.

As another aspect, the present invention provides a food composition comprising an antioxidant or antimicrobial composition containing a saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient.

The saturated butanol fraction of the butanone methanol extract is as described above, and when the composition of the present invention is used as a food additive, the saturated butanol fraction of the butanone methanol extract may be added as it is or used together with other food or food ingredients. It can be used suitably according to a conventional method. The mixed amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment), and may further include food acceptable food additives. Since the composition of the present invention is an extract derived from natural products and fractions thereof as an active ingredient, there is no problem in terms of stability because there is no problem in terms of stability.

The food composition of the present invention may include all foods having a conventional meaning, and may be mixed with terms known in the art such as functional foods and health functional foods.

The term "functional food" of the present invention means a food prepared and processed using raw materials or ingredients having a useful function to the human body according to the Act No. 6775 Act on health functional food, "functional" refers to the structure of the human body And ingestion for the purpose of obtaining nutrients for function or for obtaining useful effects in health uses such as physiological actions.

In addition, the term "health functional food" of the present invention refers to a food prepared and processed by a method of extracting, concentrating, refining, and mixing a specific ingredient as a raw material or contained in a food ingredient for the purpose of health supplement, By means of the ingredient refers to foods that are designed and processed to fully exert bioregulatory functions on the living body such as biodefense, regulation of biorhythms, prevention and recovery of diseases, and the composition for health foods prevents diseases and prevents diseases. It can perform functions related to recovery.

There is no limitation on the kind of foods in which the compositions of the present invention can be used. In addition, the composition comprising the saturated butanol fraction of the butcherson methanol extract of the present invention as an active ingredient may be prepared by mixing known additives with other suitable auxiliary ingredients that may be contained in food according to the choice of those skilled in the art. Examples of foods that can be added include meat, sausages, breads, chocolates, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products, including ice cream, various soups, beverages, teas, drinks, alcoholic beverages and Vitamin complexes, and the like, and extracts and fractions thereof according to the present invention can be prepared by adding to juice, tea, jelly and juice prepared as a main component.

In addition, foods that can be applied to the present invention include, for example, special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen noodles, noodles, etc.), health supplements , Seasoned foods (e.g. soy sauce, miso, red pepper paste, mixed soy sauce), sauces, sweets (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.) ), Beverages (e.g. fruits, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (e.g. ramen soup, etc.).

When the nutraceutical composition of the present invention is used in the form of a beverage, it may contain various sweeteners, flavoring agents or natural carbohydrates, etc. as additional ingredients, as in general beverages. In addition to the above, the nutraceutical composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin , Alcohols, carbonating agents used in carbonated drinks, and the like. Others may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks.

As another aspect, the present invention provides a pharmaceutical composition comprising an antioxidant or antimicrobial composition containing a saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient.

The saturated butanol fraction of the butanone methanol extract is as described above, and the saturated butanol fraction of the butanone methanol extract of the present invention has an antioxidant effect of eliminating free radicals (active oxygen), dermatitis, and food poisoning ( It has antimicrobial effect on microorganisms related to food poisoning and pneumonia, so it can be used as pharmaceutical composition to prevent and improve diseases caused by free radicals and microbial infectious diseases that can be caused by non-pathogenic or pathogenic microorganisms. And pharmaceuticals for treatment.

The antioxidant is as described above, and thus may be included in the pharmaceutical composition for the purpose of antioxidant, and may have a prophylactic or therapeutic effect of a disease caused by free radicals. Diseases caused by the active oxygen are not limited thereto, but degenerative neurological diseases including atherosclerosis, Lou Gehrig's disease, Parkinson's disease, Alzheimer's disease, Amyotrophic sclerosis and Huntington's disease, myocardial infarction, angina pectoris, coronary artery disease, ischemic heart Cardiovascular diseases including diseases, ischemic brain diseases including stroke, digestive system diseases including diabetes, gastritis and gastric cancer, cancer, leukemia, aging, rheumatoid arthritis, hepatitis, atopic dermatitis, and the like. Preferably may be aging caused by free radicals.

In addition, the antimicrobial is as described above, and thus may be included in the pharmaceutical composition for the purpose of antimicrobial, Staphylococcus aureus ( Staphylococcus aureus ), Staphylococcus epidermidis (Staphylococcus epidermidis) and Haemophilus influenzae (Haemophilus influenzae) and can have the same dermatitis (dermatitis), food poisoning prevention or treatment of microbial infectious diseases effects associated with (food poisoning), pneumonitis (pneumonia) caused .

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" in the present invention means to exhibit properties that are not toxic to cells or humans exposed to the composition. The carrier can be used without limitation so long as it is known in the art such as buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, bases, excipients, lubricants.

In addition, the pharmaceutical compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and the like, oral formulations, suppositories, and sterile injectable solutions, respectively, according to a conventional method. have. Furthermore, it may be used in the form of an external preparation for skin in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel or gel. Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate (calcium) in the saturated butanol fraction of the methanol extract. carbonate, sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid preparations for oral use may include various excipients, such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to water and liquid paraffin, which are commonly used to include suspensions, solutions, emulsions, and syrups. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

On the other hand, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "administration" of the present invention means introducing a predetermined substance into an individual in an appropriate manner and the route of administration of the composition can be administered via any general route as long as it can reach the target tissue. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, nasal administration, pulmonary administration, rectal administration, but is not limited thereto.

The term "individual" refers to all animals, including humans, mice, mice, livestock, and the like. Preferably, it may be a mammal including a human.

The term “pharmaceutically effective amount” means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to the medical treatment and not causing side effects, wherein the effective dose level is the sex, age and weight of the patient. Well-known factors such as health status, type of disease, severity, drug activity, drug sensitivity, method of administration, time of administration, route of administration, and rate of release, duration of treatment, combination or drug used, and other medical fields. It can be easily determined by those skilled in the art according to known factors. Dosing may include administering the recommended dose once daily, or in divided doses.

Since the saturated butanol fraction of the methanol extract of Busonson of the present invention is a natural medicinal plant as a raw material, even when used as a cosmetic composition, a food composition, or a pharmaceutical composition, the side effects may be less than that of a general synthetic compound, and thus may be included safely. .

Antioxidant or antimicrobial composition comprising Selaginella tamariscina extract of the present invention, a fraction thereof or a compound isolated therefrom as an active ingredient has an excellent activity of eliminating DPPH free radicals in a concentration-dependent manner, that is, an antioxidant effect. Has excellent growth inhibitory activity against causative organisms such as food poisoning, dermatitis, pneumonia, cellulitis, and the like. It can be usefully used in cosmetics, food and medicine for prevention, improvement or treatment.

1 is a schematic diagram illustrating a process of preparing Selaginella tamariscina methanol extract (STME) and a solvent fraction thereof according to the sequential solvent fractions.
2 is a diagram showing the results of gas chromatograph-mass spectrometer (GC / MS) analysis of the Selaginella tamariscina methanol extract (STME) and the solvent fraction thereof of the present invention, obtained according to a sequential solvent fraction.
FIG. 3 is a schematic view illustrating a process for preparing Selaginella tamariscina methanol extract (STME) and a water saturated n-butanol fraction thereof of the present invention.
Figure 4 is a diagram showing the results of gas chromatograph-mass spectrometer (GC / MS) analysis of Busonson saturated butanol fraction of the present invention.
FIG. 5 is a graph showing the concentration-dependent DPPH free radical scavenging effect of the Busonson methanol extract (STME) of the present invention and its saturated butanol fraction. Here, Asc is ascorbic acid used as a positive control group, Veh is a DMSO (dimethyl sulfoxide) treatment group which is a solvent used when dissolving the butanol saturated butanol fraction.
6 is an electrophoretic photograph showing the metal ion catalyst oxidation inhibition (BSA degradation) effect, that is, protein protection effect, according to the concentration-specific treatment of the butcherson methanol extract and the butcherin butane-saturated butanol fraction of the present invention. Here, Asc is ascorbic acid used as a positive control group, Veh is a DMSO (dimethyl sulfoxide) treatment group which is a solvent used when dissolving the butanol saturated butanol fraction.
Figure 7 is a graph showing the results of cell activity inhibition experiments according to the concentration of butcherson methanol extract and butcherson butanol saturated butanol fraction of the present invention. Here, Ctrl (control) is an untreated group, Veh is a DMSO (dimethyl sulfoxide) treated group.
Figure 8 is a graph showing the results of the cytotoxicity test (LDH assay) for the mouse splenocytes (splenocyte) according to the treatment of the concentrations of the butcherson methanol extract and Butcherson saturated butanol fraction of the present invention. Here, hydrogen peroxide (H ₂ O ₂ ) was used as a negative control group to induce strong cell inhibition, Ctrl (control) is a non-treated group, Veh is DMSO (dimethyl sulfoxide) treated group.
Figure 9 is a schematic diagram showing the acute toxicity class test flow according to oral administration of the butcherson methanol extract of the present invention.
Figure 10 is Staphylococcus aureus ( Staphylococcus aureus ) by the treatment according to the concentration of Busonson saturated butanol fraction of the present invention, Staphylococcus epidermidis (Staphylococcus epidermidis) and the a graph measured by the Haemophilus influenzae (Haemophilus influenzae) minimum inhibitory concentration for the measurement of antibacterial activity (Minimum inhibitory concentration assay).
11 is a Staphylococcus aureus by treatment of the concentration of Busonson saturated butanol fraction of the present invention by the method of measuring the minimum inhibitory concentration, Star is a photograph showing the antimicrobial activity of the pillow nose kusu epidermidis (Staphylococcus epidermidis) and Haemophilus influenzae (Haemophilus influenzae).

Hereinafter, the configuration and effects of the present invention through the embodiments will be described in more detail. These examples are only for illustrating the present invention, but the scope of the present invention is not limited by these examples.

Example 1 Preparation of Butcherson Methanol Extract and Solvent Fraction thereof

In order to secure various extraction components, 100 g of washed, dried and powdered Selaginella tamariscina was immersed in 1 L of 50% methanol and extracted three times for 6 hours to obtain a Buddha extract methanol. The methanol extract was about 10.34 It was confirmed in% yield. The obtained methanol extract was filtered and evaporated under reduced pressure to remove methanol, and the methanol fraction was evaporated to remove the methanol extract (STME; Selaginella tamariscina metanol extract). Specifically, the sequential solvent fraction is fraction extracted by adding n-hexane to n-hexane methanol extract, and methylene chloride is added to the water layer (aqueous residue) obtained in the process of obtaining the Bu-son hexane fraction. Extracted. Fraction extraction was carried out by adding ethyl acetate to the water layer obtained in the process of obtaining the methylene chloride fraction, and water-saturated n-butanol was added to the water layer obtained in the process of obtaining the ethyl acetate fraction. In addition, the water layer obtained in the process of obtaining the Busonson n-butanol fraction was also obtained (Fig. 1).

Example 2: Analysis of Active Components of Butcherson Methanol Extract and Solvent Fraction thereof

Butcherson methanol extract obtained in Example 1 and each solvent fraction layer (hexane fraction layer, methylene chloride fraction layer, ethyl acetate fraction layer, water-saturated-n-butanol (water) Gas chromatograph-mass spectrometer (GC / MS) analysis was performed to analyze the active components of the saturated n-butanol) fraction layer and the remaining water residues).

GC / MS analysis was performed using GC / MS (Agilent Technologies 5975C) equipped with a CTC CombiPAL autosampler system (Palo Alto, Calif., USA). At this time, the column used for sample analysis was an HP-5 column (HP-5 column; Agilent Technologies, 250 μm × 0.25 μm × 30 m), and the analysis conditions were held at 50 ° C. for 5 minutes and then 10 ° C. per minute. The reaction was carried out by increasing the temperature and holding at 310 ° C. for 5 minutes. All samples were prepared by GC / MS analysis with methanol extract in a 1: 1 mixture of methanol and water, hexane fraction in ethanol, methylene chloride fraction, ethyl acetate fraction and saturated butanol fraction in methanol. After dissolving at 10 mg / ㎖ using 5 μl by split injection (splitless injection).

GC / MS analysis showed that the methanol extract (STME; Selaginella tamariscina metanol extract) and its fraction (hexane fraction), methylene chloride (MC) fraction, ethyl acetate fraction and saturated-n- It was confirmed that oleic acid (9-Octadecenoic acid) and butanediol (BDO) were present in butanol (water saturated n-butanol fraction), and contained in a high content in each fraction (Fig. 2).

On the other hand, active ingredients such as oleic acid (9-Octadecenoic acid) and butanediol (BDO) contained in the butcherson methanol extract and fractions thereof are known as compounds having antimicrobial activity.

Therefore, through the above results, butsonson methanol extract and its solvent fraction (hexane fraction, hexane fraction, methylene chloride (MC) fraction, ethyl acetate fraction and ethyl saturation (n-butanol) water saturated n-butanol ) Fraction can be used as a material having an antimicrobial activity.

Example 3 Preparation of Saturated Butanol Fraction of Butcherson Methanol Extract and Establishment of Extraction Process

On the basis of the results of Example 1 and Example 2 to secure the yield by the conditions of the extraction process, it was intended to establish the extraction standard for the Buddha extract methanol extract of the mother extract of the fraction. Specifically, 100 g of Selaginella tamariscina was repeated three times for 6 hours at room temperature, 35 ° C., and 50 ° C., respectively, and at 30%, 50%, and 70% methanol concentration. Extraction was confirmed an optimal yield extraction method.

As a result, as shown in Figure 3, the solvent is 70% methanol, the extraction temperature is selected to 50 ℃, to obtain the saturated butanol solvent fraction directly from the Buddha methanol extract through the establishment of extraction standardization of the parent extract of the Buddha extract methanol An extraction process can be established. Thus, by applying the standard extraction method of 70% methanol, extraction temperature is 50 ℃ to obtain a methanol extract in 12.9% yield.

Example 4 Identification and Content Analysis of Main Components of Busonson Saturated Butanol Fractions

Gas chromatograph-mass spectrometer (Agilent Technologies 5975C) analysis was performed to analyze the major components and contents contained in the Busonson saturated butanol fraction obtained through the extraction process finally established in Example 3. Was performed.

Specifically, 100 g of Selaginella tamariscina was immersed in 1 L of 70% methanol (methanol) at 50 ° C. under extraction temperature, and extracted three times for 6 hours to obtain the Buddha extract methanol. After methanol extraction, a Butcherson saturated butanol fraction was obtained therefrom, and the yield thereof was 27.4%, which was about 3.5% of 100 g of Butcherson powder. GC / MS analysis was performed using the obtained Busonson saturated butanol fraction.

GC / MS analysis revealed that the α-linolenic acid (syn. 9,12,15-Octadecatrienoic acid), 3-dioxy-di-mannoic acid (3-Deoxy-d- mannoic acid), butanediol and guiacol (guaiacol) was found to contain (Table 1 and Figure 4). Α-linolenic acid (syn. 9,12,15-Octadecatrienoic acid), 3-deoxy-d-mannoic acid contained in the Busonson saturated butanol fraction Active ingredients such as butanediol and guiacol are known as compounds having antimicrobial and / or anti-inflammatory efficacy.

Therefore, through the results, α-linolenic acid (α-linolenic acid; syn. 9,12, 15-Octadecatrienoic acid), 3-deoxy-d-mannoic acid, butanediol ) And guaiacol (guaiacol) was found that the Busonson saturated butanol fraction can be used as a material having antimicrobial activity and anti-inflammatory effect.

No. RT
(retention time, min) Area% Library ID Qual.
(qualitative) One 31.71 11.19 2,4,6-Cycloheptatrien-1-one, 3,5-bis-trimethylsilyl- 43 2 24.26 7.78 9,12,15-Octadecatrienoic acid, methyl ester, (Z, Z, Z)- 99 3 13.07 7.37 Guaiacol 93 4 20.06 4.93 3-Deoxy-d-mannoic acid 43 5 23.46 0.37 1,2-Butanediol, 1- (2-furyl) -2,3-dimethyl- 22

Example 5: Antioxidative Effect of Busonson Methanol Extract and its Saturated Butanol Fraction

Antioxidant effect of Busonson methanol extract and its saturated butanol fraction were analyzed by DPPH radical scavenging efficacy test and metal ion catalyst oxidation inhibition test.

Example 5-1 Analysis of Antioxidant Activity of Buddha Extract Methanol Extract and its Saturated Butanol Fraction by DPPH Radical Scavenging Assay

In order to confirm the direct radical scavenging ability of Selaginella tamariscina metanol extract (STME) and its saturated butanol fraction (BuOH fraction of STME), DDPH radical scavenging efficacy test was performed. DPPH (1,1-Diphenyl-2-picrylhydrazyl) is a chemically stabilized water-soluble free radical, a purple compound that exhibits characteristic light absorption at 517 nm, and is very stable and gives off electrons when it encounters an antioxidant Radical (DPPH) disappears and the color changes from purple color to yellow color of the first solution.

Specifically, the Busonson methanol extract and its saturated butanol fractions were serially dilution from 100 μg to 1.56 μg in two-fold ratios to react with DPPH, and the absorbance at 517 nm was measured to compare the radical scavenging ability. At this time, ascorbic acid (50 μM) having an excellent antioxidant effect was used as a positive control group.

As a result of DPPH radical scavenging efficacy test, 12.5 ㎍ for Busonson methanol extract Significant radical scavenging ability was observed at the above test concentrations, and 6.25 ㎍ in case of Busonson saturated butanol fraction Concentration-dependent significant radical scavenging activity was confirmed at the above test concentrations. Specifically, 50 μg As a result of reacting the Busonson saturated butanol fraction with DPPH at the above test concentration, it was confirmed that it showed a remarkably excellent antioxidant effect (FIG. 5).

Therefore, through the above results, it was found that the antioxidant activity of the butcherson methanol butanol butanol butanol fractions separated therefrom.

Example 5-2 Analysis of Antioxidant Activity of Buddha Extract Methanol Extract and Its Saturated Butanol Fractions by Metal Ion Catalyst Protection Test

Metal ion catalyst oxidation inhibition test is a method to determine the effect of antioxidants that protect proteins or enzymes from damage caused by reactive oxygen species (ROS).

Specifically, BSA (bovine serum albumin, Sigma-Aldrich, MO, USA) was used as a target protein for verifying the antioxidant effect, and Cu ²⁺ (100 μM) and H ₂ O ₂ were used as the primary reactions. (2.5 mM) was added to generate hydroxyl radicals and then diluted with BSA and each concentration (6.25 μg, 12.5 μg, 25 μg, 50 μg, 100 μg). After mixing the fractions a second reaction was carried out. After each reaction, the experimental group was subjected to electrophoresis on 10% SDS polyacrylamide gel to inhibit the degradation of BSA protein degradation by the Buddha-methanol extract or Buddha-son saturated butanol fraction. Confirmed. At this time, ascorbic acid (50 μM) was used as a positive control group.

As a result of the metal ion catalyst oxidation inhibition test, both the Methanol extract and the Buson dehydrated butanol fractions showed a significant protein degradation protection effect even when the BSA was mixed at the lowest test concentration of 6.25 ㎍. In addition, ascorbic acid used as a positive control group was found to have an equivalent effect or more. As a result, it was confirmed that the Buddha methanol fraction and Buddha son saturated butanol fraction had a strong antioxidant effect (FIG. 6).

Thus, the results showed that the antioxidant activity of the Buddha methanol extract and Buddha son saturated butanol fraction was excellent, and that the Buddha methanol extract and Buddha son saturated butanol fraction were effective in protecting the protein from radical attack. there was.

Example 6: Toxicity Evaluation of Butcherson Methanol Extract and Its Saturated Butanol Fractions

In case of extracts derived from natural products, it is often difficult to develop the material due to strong efficacy and accompanying toxicity. Accordingly, the cell survival rate (CCK) and cytotoxicity (LDH) was confirmed to evaluate the effect of the Buddha extract methanol extract and its saturated butanol fraction on the cells. In addition, animal experiments confirmed the acute toxicity of oral administration of Buddha extract methanol extract.

Example 6-1 Cell Viability Assay by Treatment of Busonson Methanol Extract and its Saturated Butanol Fraction

In the case of extracts derived from natural products, it is often difficult to develop materials due to strong efficacy and accompanying toxicity. Therefore, cell activity tests were performed on the Buddha extract methanol extract and Buddha extract saturated butanol fraction. At this time, the cells were used as a macrophage cell line Raw 264.7 (mouse macrophage cell line RAW 264.7). The Raw 264.7 cells were adjusted to 37 ° C., 5% CO ₂ using DMEM Complete Medium containing 10% FBS (fetal bovine serum) (Dulbecco's Modified Eagle's Medium) complete medium; Hyclone, Logan, UT, USA) Incubated in a CO ₂ incubator.

Cells produce the necessary energy through the process of electron transport in the mitochondria. In this process, succinate dehydrogenase (SDH) in the electron transport system decomposes the tetrazolium salt to form insoluble formazan (formazan). Will be generated. According to this principle, cell viability was measured by measuring the amount of formazan produced using a cell counting kit-8 (CCK-8; Dojindo, Kumamoto, Japan).

Specifically, raw 264.7 cells, which are mouse macrophage cell lines, by concentration (1.56 μg / ml, 3.12 μg / ml, 6.25 μg / ml, 12.5 μg / ml, 25 μg / ml, 50 μg / ml, 100 μg / ml) After treating the methanol extract (STME) and the butanol saturated butanol fractions isolated therefrom, the mitochondrial activity of the cells was analyzed by adding CCK solution at 1:10 within 24 hours.

As a result, as compared to the hydrogen peroxide (H ₂ O ₂ ) treatment group which induced strong cell inhibition as a negative control group, even when the Busonson methanol extract and Busonson saturated butanol fraction were treated at the highest treatment concentration of 100 ㎍ / ml Since no change was observed, it was confirmed that there was no effect on the cells even at high concentrations showing strong antioxidant efficacy (FIG. 7).

Example 6-2: Cytotoxicity Evaluation of Bucherson Methanol Extract and Its Saturated Butanol Fraction (Lactate dehydrogenase (LDH) Release Assay)

In order to determine whether the cell viability of the mouse macrophage cell line, Raw 264.7, is immortalized as well as its direct cytotoxicity against primary cells, spleen cells were extracted after the mouse spleen was extracted. ) Was isolated and subjected to cytotoxicity test. Cytotoxicity test was performed to measure the amount of LDH released from damaged cells using a non-radioactive LDH assay kit, CytoTox 96 ^® (Promega, Madison, WI, USA).

The cytotoxicity test was performed in the same manner as in the cell activity assay of Example 6-1. Butcherson methanol extract and Butcherson saturated butanol fraction at the concentration of ㎖ was measured by treating the splenocytes, respectively.

Specifically, after treatment with Methanol extract and Busonson saturated butanol fractions by concentration to splenocytes extracted from mice, the level of LDH flowed out from dead cells by adding LDH solution 1: 1 within 24 hours. Was analyzed.

As a result, it was found that even when the Buddha methanol extract and Buddha's saturated butanol fraction were treated at the highest concentration of 100 ㎍ / ml, no direct cytotoxicity was induced, and the Buddha's methanol extract and Buddha's saturated butanol fraction were very low in cytotoxicity. It was confirmed (FIG. 8).

Example 6-3: Evaluation of Acute Toxicity According to Oral Administration of Buddha Son Methanol Extract

The single dose toxicity test was conducted according to the Ministry of Food and Drug Safety Guideline. The Buddha extract methanol was administered directly to the stomach by the oral administration method, and fasted for 4 hours before administration. The concentration of Methanol extract of Busonson was administered at the concentration of 300 mg / kg · bw (body weight) for the first time in accordance with the Ministry of Food and Drug Safety Guidelines. After that, it was observed for 7 days at 8 hour intervals. If there were no signs of abnormality, an additional dose of Busonson methanol extract at a concentration of 2000 mg / kg · bw was observed for 7 days, and then analyzed for abnormalities through autopsy (Fig. 9) (Cosmetic toxicity test animal replacement test method guideline). Ⅱ, Korea Food and Drug Administration, 2008; Guideline for Alternatives to Animal Toxicity Testing for Cosmetics Ⅶ, Korea Food and Drug Administration, 2015). Animal testing was performed with the approval of the Gangneung-Wonju National University Animal Care and Use Committee; Approval no .: GWNU-R2016-29.

No specific abnormality was observed for 24 hours immediately after oral administration of the Buddha extract methanol extract, and no abnormality was observed even after periodic observation for 14 days (Table 2). The change in body weight of each subject did not show any difference compared to the control group, and the intake of feed and water did not show a significant difference between the subjects and showed normal activity (Table 3). After 14 days of observation, no abnormal symptoms were observed. After euthanasia, congestion or atrophy of internal organs was confirmed, and no changes or deformations were found in all the subjects. It was confirmed that there was no organ damage because there was no difference. In addition, blood biochemical tests were performed on the Busonson methanol extract test group in order to examine whether the toxicity was not confirmed externally.

Through the results of Example 6, it can be seen that both the Buddha methanol extract and Buddha son saturated butanol fractions do not cause cytotoxicity or cytotoxicity and thus do not exhibit direct cytotoxicity. It was confirmed that there was no significant difference from the control group in the acute toxicity class test. Accordingly, the Buddha-methanol extract and Buddha-son saturated butanol fraction were verified to be a safe material without toxicity through in vitro and in vivo experiments.

Example 7: Antimicrobial Effect of Busonson Saturated Butanol Fractions

Antibacterial agents such as oleic acid (Oleic Acid; 9-Octadecenoic acid), butanediol (BDO), and guiacol as active ingredients in the Busonson methanol extract and Busonson saturated butanol fractions through Examples 2 and 4 above And / or it was confirmed that the compound having an anti-inflammatory effect, and through Example 5 and Example 6, the Buddha methanol extract and Buddha Son saturated butanol fraction has excellent anti-oxidation without toxicity to cells and laboratory animals Based on this, it was judged that it could be developed as an antimicrobial and preservative material, which has emerged as the importance of verification of toxicity. Based on this, star by treatment of the concentration of Buddha-saturated butanol fraction obtained from Buddha extract methanol Staphylococcus aureus , Staphylococcus epi pile confirmed an antimicrobial effect on the display (Staphylococcus epidermidis) and Haemophilus influenzae (Haemophilus influenzae).

Specifically, in order to confirm the direct antimicrobial effect of Budsonson saturated butanol fraction, three strains of known strains known as causative bacteria such as food poisoning, dermatitis, pneumonia, cellulitis, and Staphylococcus OY Staphylococcus aureus , Staphylococcus epi pile was confirmed by using the display (Staphylococcus epidermidis) and Haemophilus influenzae minimum inhibitory concentration assay (Minimum inhibitory concentration assay, MIC assay ) the antimicrobial activity of the (Haemophilus influenzae).

The strains used for the measurement of the minimum inhibitory concentration were prepared in advance to be 10 ⁸ CFU (colony forming unit) / mL according to the Mcfarland standard turbidity (Mcfarland standard), and the maximum concentration was 1,000 ㎍ / mL for the Busonson saturated butanol fraction. It was prepared by serial dilution to 62.5 ㎍ / ㎖ at a 2-fold ratio. The prepared three strains were inoculated at a ratio of 1: 100 as a concentration that satisfies the Mcfarland standard turbidity (Mcfarland standard), and then treated with each concentration of Busonson saturated butanol fraction and cultured to observe the antimicrobial activity.

As a result, three strains of Staphylococcus aureus , Staphylococcus aureus , Staphylococcus epidermidis a significant bacteriostatic effects against (Staphylococcus epidermidis) and Haemophilus influenzae (Haemophilus influenzae) was observed. Specifically, it was confirmed that bacterial growth was significantly inhibited for the three strains in a concentration-dependent manner when the Busonson saturated butanol fraction was treated at a concentration of 250 ~ 500 ㎍ / ㎖ or more, especially at a test concentration of 1,000 ㎍ / ㎖ or more Compared with ampicillin, a positive control group, it showed an equivalent antimicrobial effect (FIGS. 10 and 11).

Therefore, through the series of results, the Busonson saturated butanol fraction extracted directly from Busonson methanol extract without performing sequential solvent fractionation, including strong reactive oxygen species (ROS) scavenging ability and protein protection, Staphylococcus aureus ( Staphylococcus aureus ), Staphylococcus epidermidis (Staphylococcus epidermidis) and Haemophilus influenzae has been verified that the antibacterial effect of the (Haemophilus influenzae), can be seen that useful as a safe natural materials that can replace synthetic preservatives and antimicrobial The there was.

Preparation Example 1 Preparation of Antioxidant or Antimicrobial Cosmetics Containing Saturated Butanol Fraction of Butcherson Methanol Extract as an Active Ingredient

Preparation Example 1-1 Preparation of Flexible Lotion

A flexible lotion containing the saturated butanol fraction of the butcherson methanol extract as an active ingredient was prepared as shown in Table 5 below.

Raw material Content (parts by weight) Saturated Butanol Fraction of Butcherson Methanol Extract of the Present Invention 10.00 1,3-butylene glycol 1.00 Disodium ID 0.05 Allantoin 0.10 Dipotassium glycylizate 0.05 Citrix Acid 0.01 Sodium citrate 0.02 Glyceres-26 1.00 Arbutin 2.00 Hydrogenated Castor Oil 1.00 ethanol 30.00 Preservative a very small amount coloring agent a very small amount Flavor a very small amount Purified water Remaining amount

Preparation Example 1-2: Preparation of Nutritional Cream

* The flexible nutrition cream containing the saturated butanol fraction of the butcherson methanol extract as an active ingredient was prepared as shown in Table 6.

Raw material Content (parts by weight) Saturated Butanol Fraction of Butcherson Methanol Extract of the Present Invention 10.0 1,3-butylene glycol 7.0 glycerin 1.0 D-panthenol 0.1 Plant extracts 3.2 Magnesium Aluminum Silicate 0.3 PEG-40 Stearate 1.2 Stearic acid 2.0 Polysorbate 60 1.5 Lipophilic glyceryl stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 3.0 Mineral oil 4.0 Squalane 3.8 Carlylic / Capric Triglycerides 2.8 vegetable oil 1.8 Dimethicone 0.4 Dipotassium glycylizate a very small amount Allantoyl a very small amount Sodium hyaluronate a very small amount Tocopheryl Acetate Quantity Triethanolamine Quantity Preservative Quantity Flavor Quantity Purified water Remaining amount

Preparation Example 2 Preparation of Antioxidant or Antimicrobial Foods Containing Saturated Butanol Fraction of Butcherson Methanol Extract as an Active Ingredient

Preparation Example 2-1 Preparation of Wheat Flour Food

0.5 to 5.0 parts by weight of the saturated butanol fraction of the butcherson methanol extract of the present invention was added to the flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture.

Preparation Example 2-2: Preparation of Soup and Gravy

0.1 to 5.0 parts by weight of the saturated butanol fraction of the butcherson methanol extract of the present invention was added to soups and broths to prepare meat products for health promotion, soups and noodles for noodles.

Preparation Example 2-3 Preparation of Ground Beef

10 parts by weight of the saturated butanol fraction of the butcherson methanol extract of the present invention was added to the ground beef to prepare a ground beef for health promotion.

Preparation Example 2-4: Preparation of Dairy Products

5-10 parts by weight of the saturated butanol fraction of the butcherson methanol extract of the present invention was added to milk to prepare various dairy products such as butter and ice cream.

Preparation Example 2-5: Preparation of Wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh.

Black beans, black sesame seeds, and perilla were also steamed and dried in a known manner, and then roasted to prepare a powder having a particle size of 60 mesh.

The saturated butanol fractions of the methanol extract of butcherone of the present invention were concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by spraying and drying with a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The grains, seeds and the saturated butanol fraction of the methanol extract of the present invention prepared above were prepared by combining the following ratios:

Cereals (30 parts by weight brown rice, 15 parts by weight of barley, 20 parts by weight of barley), seed seeds (7 parts by weight perilla, 8 parts by weight of black soybeans, 7 parts by weight of black sesame seeds), saturated butanol fraction of the methanol extract of the present invention 3 parts by weight), ganoderma lucidum (0.5 parts by weight), turmeric (0.5 parts by weight).

Preparation Example 2-6 Preparation of Health Beverage

Homogeneously add 5 g of the saturated butanol fraction of the Buddha extract of the present invention and subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%). After sterilization by mixing, it was prepared by packaging in a small packaging container such as glass bottles, plastic bottles.

Preparation Example 2-7: Preparation of Vegetable Juice

Vegetable juice was prepared by adding 5 g of the saturated butanol fraction of the Busonson methanol extract of the present invention to 1,000 ml of tomato or carrot juice.

Preparation Example 2-8 Preparation of Fruit Juice

Fruit juice was prepared by adding 1 g of the saturated butanol fraction of the Busonson methanol extract of the present invention to 1,000 ml of apple or grape juice.

Preparation Example 3 Preparation of Antioxidant or Antimicrobial Pharmaceutical Composition Containing Saturated Butanol Fraction of Butcherson Methanol Extract as an Active Ingredient

Preparation Example 3-1 Preparation of Powder

1 g of lactose was mixed with 2 g of the saturated butanol fraction of the butanone methanol extract of the present invention and filled into an airtight cloth to prepare a powder.

Preparation Example 3-2: Preparation of Tablet

Tablets were prepared by mixing 100 mg of the saturated butanol fraction of the Busonson methanol extract of the present invention, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate, followed by compression according to a conventional method for preparing tablets.

Preparation Example 3-3 Preparation of Capsule

100 mg of saturated butanol fraction of the Buddha extract methanol of the present invention, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate are mixed and then filled into gelatin capsules according to a conventional method for preparing capsules to prepare capsules. It was.

Preparation Example 3-4: Preparation of Ring

1 g of the saturated butanol fraction of the butcherson methanol extract of the present invention, 1.5 g of lactose, 1 g of glycerin, and 0.5 g of xylitol were mixed and prepared to be 4 g per ring according to a conventional method.

Preparation Example 3-5: Preparation of Granules

After mixing the saturated butanol fraction 150 mg, soybean extract 50 mg, glucose 200 mg and starch 600 mg of the methanol extract of the present invention, dried 100 ° C. with 30 mg of ethanol and dried at 60 ℃ to form granules Filled in the gun.

Claims (3)

A pharmaceutical composition for preventing or treating pneumonia caused by Haemophilus influenzae, which contains a saturated butanol fraction of Selaginella tamariscina methanol extract as an active ingredient.
According to claim 1, wherein the saturated butanol fraction of the Buddha extract methanol is α-linolenic acid (α-linolenic acid), 3-deoxy-d-mannoic acid (3-Deoxy-d-mannoic acid), guaiacol) and butanediol compounds.
The compound according to claim 2, wherein the compound has 5 to 10 parts by weight of α-linolenic acid, 3-dioxy-di-mannonic acid, based on 100 parts by weight of the saturated butanol fraction of the Buddha extract methanol. 3-Deoxy-d-mannoic acid) 3 to 6 parts by weight, guaiacol (guaiacol) 5 to 10 parts by weight, and butanediol (butanediol) is characterized in that it comprises 0.1 to 3 parts by weight Composition.

Images