KR20220094309A - Pharmaceutical composition for preventing or treating infectious symptoms by pathogenic and apathogenic bacteria comprising Oenothera biennis extract as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising an Oenothera biennis extract as an active ingredient for preventing or treating infectious diseases caused by pathogenic and apathogenic bacteria. Specifically, it was confirmed that the Oenothera biennis extract exhibits an antibacterial activity that inhibits the proliferation of pathogenic bacteria such as Escherichia coli O157:H7 and pyogenic bacteria (Staphylococcus aureus), and apathogenic bacteria such as Escherichia coli DH5α and acne causing bacteria (Propionibacterium acnes), and also exhibits the ability to inhibit the growth of the bacteria and kill the same in a concentration-dependent manner. Accordingly, the Oenothera biennis extract can be effectively used as an active ingredient of a composition for inhibiting the growth of pathogenic and apathogenic bacteria, an antibacterial composition, and a composition for preventing, alleviating, or treating infectious diseases caused by the pathogenic and the apathogenic bacteria.

Description

A pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, comprising an evening primrose extract as an active ingredient.

The present invention relates to a pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria comprising an evening primrose ( Oenothera biennis ) extract as an active ingredient, and more particularly, an evening primrose extract having the ability to inhibit and kill pathogenic and non-pathogenic bacteria It relates to a pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, including as an active ingredient.

Pathogenic microorganisms cause several infectious diseases in humans and livestock. As an example, Gram-positive bacteria, Staphylococcus aureus , are one of the causative bacteria that invade the human body and cause skin infections such as abscesses or wound infections, osteoarthritis, bacteremia, pneumonia, toxin shock syndrome, and food poisoning. Aeruginosa is a causative agent that causes infectious diseases in animals, and in particular, Escherichia coli causes acute mastitis in cattle with Escherichia coli. Escherichia coli belongs to Gram-negative bacteria, and is generally present in the intestinal tract of humans or livestock, but is also detected in the urinary tract or respiratory tract in animals. E. coli O157:H7 ( E coli 0157:H7 ), a representative pathogenic E. coli, is Enterohemorrhagic Escherichia coli and causes food poisoning, hemolytic uremic syndrome, hemolytic colitis, and renal dysfunction in humans. do. In addition, it has a very low ID ₅₀ , so the infection rate is very high.

In addition, the causative agent of acne (Propionibacterium acnes) causes non-pathogenic bacteria or inflammatory skin diseases. For example, acne is a chronic inflammatory disease of the hair follicles and sebaceous glands that mainly occurs during puberty, and is a disease characterized by the formation of comedones, papules and nodules. Although the cause is not certain, it has been found that the most important factors are androgens that stimulate the sebaceous glands and acne-causing bacteria that break down sebum and produce free fatty acids while breeding in the fur glands. , When the follicle wall cells become keratinized and the follicle follicles are blocked, sebum cannot be discharged and stagnate, the bacteria causing acne breed here.

On the other hand, in general, direct or indirect damage caused by pathogenic microorganisms causes many economic, environmental and medical problems. In particular, the loss due to spoilage during the food distribution process in the food industry, the harmfulness and environmental pollution to the human body due to the use of excessive chemical pesticides on crops in agriculture, and the emergence of antibiotic-resistant strains due to the misuse of antibiotics have already occurred in society. It is becoming a chronic problem in general. In this reality, natural physiologically active substances isolated from plants and the like exhibit antibiotic activity in a wide range of pathogenic bacteria and fungi, but development as a new alternative material is urgent in that they can be used environmentally friendly.

Evening primrose is a dicotyledonous plant, a biennial plant of the genus oleracea, native to South America, and distributed as a naturalized plant in North America, Korea, Japan, and China. The medicinal evening primroses used include Oenothera odorata , Oenothera biennis , and Oenothera brevipes , which have different efficacy and medicinal properties.

In oriental medicine, the root of the evening primrose is used as a medicinal herb called moongyeoncho (月見草), and when the fever is high due to a cold, put it in water and take it as a decoction.

Accordingly, the present inventors have made efforts to develop a natural material having no side effects, high safety, and excellent antibacterial efficacy against pathogenic bacteria. Staphylococcus aureus ), a non-pathogenic bacterium Escherichia coli DH5α ( Escherichia coli DH5α ), and a bacterial growth curve of Propionibacterium acnes ) Paper disc diffusion assay, microplate analysis, and colony suppression comparison It was confirmed through analysis (CFU assay). Accordingly, the present invention was completed by revealing that the evening primrose extract can be very usefully used for the prevention or treatment of diseases caused by the proliferation of pathogenic and non-pathogenic bacteria and can be used as a functional material.

Republic of Korea Patent Publication No. 10-2014-0022571 Republic of Korea Patent Publication No. 10-2017-0072052 Republic of Korea Patent Publication No. 10-2010-0102850

It is an object of the present invention to provide a composition for preventing, improving or treating infections caused by pathogenic and non-pathogenic bacteria, comprising an evening primrose ( Oenothera biennis ) extract having the ability to inhibit proliferation and kill pathogenic and non-pathogenic bacteria as an active ingredient.

Another object of the present invention is to provide a composition for inhibiting the growth of pathogenic and non-pathogenic bacteria, comprising an evening primrose ( Oenothera biennis ) extract as an active ingredient.

Another object of the present invention is to provide an antibacterial composition for pathogenic and non-pathogenic bacteria, comprising an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, comprising an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides a food composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides an external preparation for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides a cosmetic composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides a composition for inhibiting the growth of pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides an antibacterial composition for pathogenic and non-pathogenic bacteria containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

Evening primrose ( Oenothera biennis ) extract according to the present invention contains pathogenic bacteria Escherichia coli O157:H7 and Staphylococcus aureus , non-pathogenic bacteria Escherichia coli DH5α ( Escherichia coli DH5α ) and acne causative bacteria ( Propionibacterium acnes ) Because it exhibits an antibacterial action to inhibit proliferation, and inhibits and kills the growth of the pathogenic and non-pathogenic bacteria in a concentration-dependent manner, the evening primrose extract is a composition for inhibiting the growth of pathogenic and non-pathogenic bacteria, an antibacterial composition, and a pathogenic and non-pathogenic It can be usefully used as an active ingredient in a composition for preventing, improving or treating infections caused by bacteria.

1 shows the pathogenic bacteria E. coli O157:H7 ( Escherichia coli O157:H7 ) and the non-pathogenic bacteria Escherichia coli DH5α ( Escherichia coli DH5α ) and the antibacterial ability against Propionibacterium acnes were observed through paper disc diffusion analysis.
2a to 2c are E. coli O157:H7 (FIG. 2A), E. coli DH5α (FIG. 2B) and pyogenic bacteria (FIG. 2c) by concentration of the evening primrose ( Oenothera biennis ) extract (EVP-EtOH) prepared in an embodiment of the present invention. It showed the effect of inhibiting the proliferation of growth.
3 shows the degree of inhibition of bacterial growth activity against E. coli O157:H7, suppurative bacteria, E. coli DH5α and acne-causing bacteria of the evening primrose ( Oenothera biennis ) extract (EVP) prepared in an embodiment of the present invention. analyzed and presented.
4A to 4D are E. coli O157:H7 (FIG. 4A), pyogenic bacteria (FIG. 4B), E. coli DH5α (FIG. 4c) and acne of the evening primrose ( Oenothera biennis ) extract (EVP-EtOH) prepared in an embodiment of the present invention. It is shown by observing the bacterial community formation for the causative bacteria (FIG. 4d).

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

In the present invention, the term "prevention" refers to inhibiting the occurrence of a disease or disease in an individual who has never been diagnosed as possessing a disease or disease, but is prone to such disease or disease. As used herein, the term “treatment” refers to (i) inhibiting a disease or development of a disease; (ii) alleviation of the disease or condition; and (iii) disease or disease elimination.

The present invention provides a pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

Evening primrose ( Oenothera biennis ) extract, which is the active ingredient of the present invention, is preferably prepared by a method comprising the following steps, but is not limited thereto:

1) extracting by adding an extraction solvent to evening primrose ( Oenothera biennis ); and

2) filtering the extract of step 1).

In the manufacturing method of the present invention, the evening primrose in step 1) may be cultivated or commercially available without limitation. In addition, for the evening primrose extract, it is preferable to use one or more parts selected from the group consisting of flowers, leaves, branches, fruits, roots, stems and seeds of evening primrose, and it is more preferable to use the stem of evening primrose, but is not limited thereto. .

In the preparation method of the present invention, it is preferable to use water, alcohol, or a mixture thereof as the extraction solvent in step 1). As the alcohol, it is preferable to use a lower alcohol of C ₁ to C ₅ , and as the lower alcohol, methanol, ethanol, butanol, and the like may be used. In addition, organic solvents such as ethyl acetate, acetone, ether, chloroform, benzene, hexane, and dichloromethane may be used. More specifically, it is preferable to use methanol or ethanol, and it is most preferable to use ethanol. As the extraction method, it is preferable to use ultrasonic extraction, shaking extraction, Soxhelt extraction, or reflux extraction, but is not limited thereto. It is preferable to wash the extraction solvent and add 1 to 15 times the amount of well-dried evening primrose for extraction, and more preferably add 2 to 10 times to extract. The extraction temperature is preferably 20 to 110 ℃, but is not limited thereto. In addition, the extraction time is preferably 10 minutes to 72 hours, more preferably 20 minutes to 48 hours, but is not limited thereto. In addition, the number of extractions is preferably 1 to 5 times, but is not limited thereto.

In the manufacturing method of the present invention, after step 2), the following steps may be additionally included:

3) concentrating the filtrate of step 2) under reduced pressure; and

4) drying the concentrate of step 3).

In the manufacturing method of the present invention, the vacuum concentration in step 3) is preferably performed using a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, drying under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying is preferable, but is not limited thereto.

In the present invention, the evening primrose extract may be included in a concentration of 10 μg/ml to 100 mg/ml, specifically 50 μg/ml to 100 mg/ml, and more specifically 100 μg/ml to 100 mg/ml It may be included at a concentration of 10 mg/ml, but is not limited thereto.

In the present invention, the pathogenic bacteria are preferably Escherichia coli O157:H7 ( Escherichia coli O157:H7 ) or Staphylococcus aureus .

In the present invention, the non-pathogenic bacteria are preferably Escherichia coli DH5α ( Escherichia coli DH5α ) or Propionibacterium acnes .

In the present invention, the infection caused by pathogenic and non-pathogenic bacteria is E. coli O157:H7 ( Escherichia coli O157:H7 ) or Staphylococcus aureus as pathogenic bacteria, Escherichia coli DH5α ( Escherichia coli DH5α ) or Propionibacterium acnes as non-pathogenic bacteria ), such as conjunctivitis, abscess, dermatitis, acne, enteritis, vaginitis, hemolytic uremic syndrome, hemolytic colitis, osteoarthritis, peritonitis, pneumonia, osteomyelitis, cellulitis, meningitis, nephritis, carditis, gastritis, esophagitis, duodenitis, Colitis, bacteremia, pneumonia, toxin shock syndrome, food poisoning, high fever, sepsis, acute breast cancer, edema disease, mastitis, enterobacteriaceae, kidney dysfunction, constipation, inflammatory bowel disease, indigestion, gas reflux, abdominal distension, gas, failure to recover It may be nausea, pain, diarrhea, irritable bowel syndrome, or a feeling of residual stool, but is not limited thereto.

In a specific embodiment of the present invention, the present inventors prepared an evening primrose ( Oenothera biennis ) extract, and the evening primrose extract is a pathogenic bacterium Escherichia coli O157:H7 ) and Staphylococcus aureus , a non-pathogenic bacterium Escherichia coli The concentration-dependent inhibition of bacterial growth in the bacterial growth curves of DH5α ( Escherichia coli DH5α ) and acne causative bacteria ( Paper disc diffusion assay), microplate analysis, and comparative analysis of colony suppression (CFU assay) were performed. confirmed through.

Therefore, the present inventors confirmed that the evening primrose extract according to the present invention exhibits an antibacterial action for inhibiting the growth of the pathogenic and non-pathogenic bacteria, and inhibits and kills the growth of the pathogenic and non-pathogenic bacteria in a concentration-dependent manner. Evening primrose extract can be usefully used as an active ingredient in a pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions.

The pharmaceutical composition according to the present invention can be administered orally or parenterally, and when administered parenterally, it is preferable to select an injection method for external application or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection. Preferably, it is not limited thereto.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions according to conventional methods, respectively. can Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the mixed herbal medicine, for example, starch, calcium carbonate, sucrose ) or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, for example, wetting agents, sweeteners, fragrances, preservatives, etc. may be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used. In addition, other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed.

The preferred dosage of the pharmaceutical composition according to the present invention varies depending on the absorption of the active ingredient in the body, the age, sex, and the degree of obesity of the patient, but may be appropriately selected by those skilled in the art. However, for desirable effects, in the case of oral administration, the composition of the present invention per 1 kg of body weight per day is generally administered to adults at 0.0001 to 100 mg/kg, preferably 0.001 to 100 mg/kg. have. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.

In addition, the present invention provides a health functional food composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides a food composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In the present invention, the evening primrose extract preferably contains 0.0001 to 100% by weight, but is not limited thereto.

In addition, the extraction method and extraction target of the evening primrose extract, pathogenic and non-pathogenic bacteria and diseases caused by them are the same as those described in the pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, containing the evening primrose extract as an active ingredient. , The specific description will refer to the above, and below, only the specific composition of the health functional food composition or food composition will be described.

On the other hand, the present inventors found that the evening primrose extract according to the present invention exhibits an antibacterial action that inhibits the growth of pathogenic bacteria E. coli O157:H7 and aeruginosa, non-pathogenic E. coli DH5α and acne causative bacteria, and concentration-dependently inhibits the growth of the bacteria And since it was confirmed that the killing ability, the evening primrose extract can be usefully used as an active ingredient in a health functional food composition or food composition for preventing or improving infection caused by the pathogenic and non-pathogenic bacteria.

The health functional food composition of the present invention refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, 'functionality' refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body. The health functional food composition of the present invention can be prepared by a method commonly used in the conventional technical field, and during the production, it can be prepared by adding raw materials and components commonly added in the conventional technical field.

In addition, the dosage form of the health functional food composition may be prepared without limitation as long as it is a dosage form recognized as a health functional food. The food composition of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage that there are no side effects that may occur during long-term administration of the drug using food as a raw material, and excellent portability to enhance the effect It can be taken as a supplement for For example, the health functional food composition may be food in the form of tablets, capsules, pills, or liquids, and the food composition includes various drinks, meat, sausages, breads, candies, snacks, noodles, ice cream, dairy products, and soups. , ion beverages, beverages, gum, tea, and vitamin complexes may be selected.

When the composition of the present invention is used as a health functional food composition or food composition, the extract of the present invention may be added as it is or used with other foods or food ingredients, and may be appropriately used according to a conventional method. In addition to the active ingredient, the composition may include a food additive that is pharmaceutically acceptable, and the mixing amount of the active ingredient may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).

The food supplement additive of the present invention refers to a component that can be supplementally added to food, and is added to prepare a health functional food composition or food composition of each formulation, and those skilled in the art can appropriately select and use it.

Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention. In addition, the extract of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

In addition, the present invention provides an external preparation for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides a cosmetic composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In the present invention, the extraction method and extraction target of the evening primrose extract, pathogenic and non-pathogenic bacteria and diseases caused by them are described in the pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, containing the evening primrose extract as an active ingredient, and Since it is the same, the specific description will refer to the above, and below, only the specific composition of the external preparation or cosmetic composition will be described.

On the other hand, the present inventors found that the evening primrose extract according to the present invention exhibits an antibacterial action that inhibits the growth of pathogenic bacteria E. coli O157:H7 and aeruginosa, non-pathogenic E. coli DH5α and acne causative bacteria, and concentration-dependently inhibits the growth of the bacteria and killing ability, the evening primrose extract can be usefully used as an active ingredient in an external preparation or cosmetic composition for preventing or improving infections caused by the pathogenic and non-pathogenic bacteria.

Components included in the external application or cosmetic composition of the present invention may include components commonly used in external preparations or cosmetic compositions in addition to the active ingredient, for example, antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances. phosphorus adjuvant, and a carrier.

In order to formulate the active ingredient of the present invention, it can be easily formulated if carried out according to a conventional method, and surfactants, excipients, colorants, spices, preservatives, stabilizers, buffers, suspending agents, and other compatibilizing adjuvants can be appropriately used.

The composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, It may be formulated as oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream, spray, and the like, but is not limited thereto. More specifically, it may be prepared in the form of softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol fatty ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, and microcrystals Sung cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

In addition, the present invention provides a composition for inhibiting the growth of pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In addition, the present invention provides an antibacterial composition for pathogenic and non-pathogenic bacteria containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.

In the present invention, the composition can be widely used as an antibacterial application in various industrial fields for blocking bacterial infection. For example, the industrial field, a pharmaceutical composition for the prevention or treatment of bacterial diseases; health functional food for the prevention or improvement of bacterial diseases; livestock feed additives for the prevention of bacterial diseases; industrial disinfectants to prevent bacterial infection; Industrial preservatives that can secure a preservation period by blocking contamination of objects from bacterial infection; Antibacterial household items such as office supplies, kitchenware, and toiletries; and antibacterial food or pharmaceutical additives, but is not limited thereto.

In the present invention, the extraction method and extraction target of the evening primrose extract and the pathogenic and non-pathogenic bacteria and diseases caused by them are described in the pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, containing the evening primrose extract as an active ingredient, and Since they are the same, the detailed description uses the above content. In addition, specific descriptions of some of the uses will refer to the above content, and only the specific configuration of the uses not disclosed above will be described below.

On the other hand, the present inventors found that the evening primrose extract according to the present invention exhibits an antibacterial action that inhibits the growth of pathogenic bacteria E. coli O157:H7 and aeruginosa, non-pathogenic E. coli DH5α and acne causative bacteria, and concentration-dependently inhibits the growth of the bacteria and killing ability, the evening primrose extract can be usefully used as an active ingredient in the composition for inhibiting the growth of pathogenic and non-pathogenic bacteria or antibacterial composition.

When the active ingredient of the present invention is used as a livestock feed additive, it may be applied to livestock feed in an appropriate amount depending on the type of livestock to be applied, bacterial infection concerns, and environmental factors, and preferably in an amount of 0.1 to 10% by weight in the feed. may be added.

The livestock may include, but is not limited to, cattle, pigs, chickens, goats, ducks, sheep, turkeys, ostriches, dogs, and cats.

When the active ingredient of the present invention is used as an industrial disinfectant, it is for preventing bacterial infection of a subject, for example, manufacturing, medical industry, livestock industry, and the like. The amount may be different depending on the purpose of use, but for example, 100 parts by weight of the active ingredient powder of the present invention may be mixed with 100 to 100,000 parts by weight of distilled water.

When the active ingredient of the present invention is used as an industrial preservative, the preservative includes various industrial instruments, devices, equipment, etc., and the amount used may be different depending on the purpose of use, for example, 100 parts by weight of the active ingredient powder of the present invention in distilled water It can be used by mixing with 100 to 100,000 parts by weight.

When the active ingredient of the present invention is used for antibacterial household products, the antibacterial household products may be household products such as kitchen cleaning, bathroom cleaning, and washing, but is not limited thereto. In addition, although the active ingredient of the present invention may be different depending on the purpose of use, it may preferably be included in an amount of 0.1 to 20% by weight of the antibacterial household products.

When the active ingredient of the present invention is used as an antibacterial food or drug additive, the active ingredient of the present invention may be different depending on the purpose of use, but may preferably be included in an amount of 0.1 to 10% by weight of the antimicrobial food or drug.

Hereinafter, the present invention will be described in detail by way of Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

<Example 1> Evening primrose ( Oenothera biennis ) preparation of extracts

After adding 150 ml of 99% ethanol to 50 g of evening primrose ( Oenothera biennis ) stems, performing an extraction reaction at 150 rpm for 48 hours, filtration, concentration under reduced pressure and freeze-drying to prepare an evening primrose extract.

<Experimental Example 1> Pathogenic and non-pathogenic bacteria culture

E. coli O157:H7 ( E. coli O157:H7 ) as a pathogenic bacterium and E. coli DH5α ( E. coli DH5α ) as a non-pathogenic bacterium were each purchased from ATCC (American type culture collection), and LB broth (Luria-Bertani broth) was used. Bacteria were activated by diluting 1:100 and culturing. Maintained and cultured under atmospheric conditions at 37°C.

As a pathogenic bacterium, S. aureus was diluted 1:100 in TSB liquid medium (trypticase soy broth) and cultured to activate the bacteria.

As a non-pathogenic bacteria, the acne causative bacteria ( P. Acnes ) is It was diluted 1:100 in RCM liquid medium (Reinforced clostridial medium), and since it is anaerobic, it was cultured in an airtight container to activate the bacteria. Maintained and cultured under atmospheric conditions at 37°C.

<Experimental Example 2> Disc sensitivity evaluation of evening primrose extract

The disc sensitivity evaluation of the evening primrose extract prepared in <Example 1> to the pathogenic and non-pathogenic bacteria of <Experimental Example 1> was evaluated using a paper disc diffusion analysis method.

Specifically, for evaluation of disk sensitivity to E. coli O157:H7 and E. coli DH5α, a 10 cm ² plastic Petri dish was solidified with LB medium containing agar agar (1.5%) and dried. The dried LB medium was treated with 100 μl of the LB medium containing E. coli O157:H7 cultured in <Experimental Example 1>, injected into a Petri dish, and spread widely with a spreader.

In addition, for the evaluation of disk sensitivity to suppurative bacteria, a TSB medium containing agar agar (1.5%) in a 10 cm ² plastic Petri dish was solidified and dried. 100 μl of the TSB medium containing the aeruginosa cultured in <Experimental Example 1> was treated in the dried TSB medium, injected into a Petri dish, and spread widely with a spreader.

In addition, for the evaluation of the disc sensitivity to the acne-causing bacteria, the RCM medium containing agar agar (1.5%) in a 10 cm ² plastic Petri dish was solidified and dried. 100 μl of the RCM medium containing the acne-causing bacteria cultured in <Experimental Example 1> was treated in the dried RCM medium, injected into a Petri dish, and spread widely with a spreader.

After smearing each of the E. coli O157:H7, Escherichia coli DH5α, aeruginosa and acne causative bacteria, 8 mm of a paper disk was placed as a sterilized fixture on the medium on which the bacteria were smeared, and then 100 mg/ After 20 μl treatment in ml concentration, it was cultured for 24 hours in a bacteria-only incubator at 37°C. After incubation, a part (clear zone) where bacteria did not grow including the paper disk was confirmed.

As a result, as shown in FIG. 1 , the clear zone was confirmed in the group with evening primrose extract for all Petri dishes smeared with E. coli O157:H7, E. coli DH5α, aeruginosa and acne causative bacteria, respectively.

From the above results, it can be seen that the evening primrose extract of the present invention has antibacterial activity against pathogenic bacteria E. coli O157:H7 and aeruginosa, non-pathogenic bacteria E. coli DH5α and acne causative bacteria.

<Experimental Example 3> Confirmation of the bacterial growth curve proliferation inhibitory effect of evening primrose extract

<3-1> Confirmation of proliferation inhibitory effect of evening primrose extract in E. coli O157:H7 growth

Evening primrose extract E. coli O157:H7 In order to examine the effect on the growth, the evening primrose extract was treated with E. coli O157:H7 by concentration, and the growth was measured.

Specifically, after diluting E. coli O157:H7 cultured in <Experimental Example 1> in LB medium, the evening primrose extract prepared in <Example 1> was added to 0 μg/ml, 100 μg/ml, 1 mg/ml, It was treated with a concentration of 2 mg/ml and the total volume was adjusted to 20 ml. Then, while culturing for 24 hours at 37°C in dry aerobic conditions, the OD value was measured with a microplate reader at a wavelength of 600 nm every 20 minutes to 1 hour.

As a result, as shown in FIG. 2a, in the group not treated with the evening primrose extract, it was confirmed that bacterial growth progressed through the logarithmic phase and the stationary phase over time. On the other hand, when the evening primrose extract was treated together, it was confirmed that the proliferation of E. coli O157:H7 was reduced. In addition, it was confirmed that the proliferation of E. coli O157:H7 decreased in a concentration-dependent manner as the concentration of the evening primrose extract was increased.

From the above results, it can be seen that the evening primrose extract of the present invention exhibits an inhibitory effect on the growth of E. coli O157:H7 in a concentration-dependent manner.

<3-2> Confirmation of proliferation inhibitory effect of evening primrose extract in the growth of E. coli DH5α

To investigate the effect of the evening primrose extract on the growth of E. coli DH5α, the evening primrose extract was treated with E. coli DH5α by concentration and growth was measured.

Specifically, OD values were measured in the same manner as in Experimental Example <3-1>, except that Escherichia coli DH5α cultured in <Experimental Example 1> was used as the bacterium.

As a result, as shown in FIG. 2b, in the group not treated with the evening primrose extract, it was confirmed that bacterial growth progressed through the logarithmic phase and the stationary phase over time. On the other hand, it was confirmed that the proliferation of E. coli DH5α was decreased when the evening primrose extract was treated together. In addition, it was confirmed that the proliferation of E. coli DH5α decreased in a concentration-dependent manner as the concentration of the evening primrose extract was increased.

From the above results, it can be seen that the evening primrose extract of the present invention exhibits an inhibitory effect on the growth of E. coli DH5α in a concentration-dependent manner.

<3-3> Confirmation of proliferation inhibitory effect of evening primrose extract in the growth of suppurative bacteria

In order to investigate the effect of the evening primrose extract on the growth of S. aeruginosa, the evening primrose extract was treated with S. aeruginosa by concentration and the growth was measured.

Specifically, the OD value was measured in the same manner as in Experimental Example <3-1>, except that the bacteria cultured in <Experimental Example 1> was used and a TSB medium was used instead of the LB medium.

As a result, as shown in FIG. 2c, in the group not treated with the evening primrose extract, it was confirmed that bacterial growth progressed through the logarithmic phase and the stationary phase over time. On the other hand, when the evening primrose extract was treated together, it was confirmed that the proliferation of suppurative bacteria was reduced. In addition, it was confirmed that as the concentration of the evening primrose extract increased, the proliferation of suppurative bacteria decreased in a concentration-dependent manner.

From the above results, it can be seen that the evening primrose extract of the present invention exhibits an inhibitory effect on the growth of suppurative bacteria in a concentration-dependent manner.

<Experimental Example 4> Confirmation of bacterial colonization inhibitory effect of evening primrose extract in growth stagnant state

In order to examine the effect of the evening primrose extract on the colony formation of pathogenic and non-pathogenic bacteria of <Experimental Example 1>, a CFU assay was performed.

Specifically, the evening primrose extract prepared in <Example 1> in the same manner as in the method described in <Experimental Example 3> was added at a concentration of 0 and 2 mg/ml to the pathogenic and non-pathogenic bacteria of <Experimental Example 1>, respectively. After treatment and culturing, the bacteria entering the plateau phase of the growth curve were diluted decimal. Thereafter, treatment and plating in Petri dishes with solidified LB medium, TSB medium or RCM medium, and colony growth was induced through culture for 24 hours. Then, the number of colonies at each dilution was compared to evaluate how inhibited the growth of bacteria was.

As a result, as shown in FIGS. 3 and 4A to 4D , it was confirmed that E. coli O157:H7, E. coli DH5α, aeruginosa and acne causative bacteria were all significantly reduced in the number of colonies in the treated group compared to the evening primrose extract untreated group.

Through the above results, it can be seen that the evening primrose extract of the present invention exhibits an effect of significantly reducing the number of colonies by inhibiting the growth of pathogenic bacteria E. coli O157:H7 and aeruginosa, non-pathogenic E. coli DH5α and acne causative bacteria.

Therefore, the evening primrose extract of the present invention exhibits an antibacterial effect through the inhibition of the life cycle of pathogenic bacteria E. coli O157:H7 and suppurative bacteria, non-pathogenic bacteria E. coli DH5α and acne causative bacteria, and pathogenic bacteria such as E. coli O157:H7 and aeruginosa and Escherichia coli It can be usefully used for the prevention or treatment of infections caused by non-pathogenic bacteria such as DH5α and acne-causing bacteria.

Preparation examples for the composition of the present invention are exemplified below. The following preparation examples are provided to help the understanding of the practice of the present invention, but do not mean that the preparation method of the formulation according to the present invention is limited to the following preparation examples.

<Preparation Example 1> The extract of the present invention Preparation of pharmaceutical preparations containing as active ingredients

<1-1> Preparation of powder

10 mg of extract of the present invention

1 g lactose

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

<1-2> Preparation of tablets

0.1 mg of extract of the present invention

100 mg cornstarch

100 mg lactose

2 mg magnesium stearate

After mixing the above ingredients, tablets were prepared by tableting according to a conventional manufacturing method of tablets.

<1-3> Preparation of capsules

0.1 mg of extract of the present invention

100 mg cornstarch

100 mg lactose

2 mg magnesium stearate

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

<1-4> Preparation of the ring

1 mg of extract of the present invention

1.5 g lactose

1 g of glycerin

0.5 g of xylitol

After mixing the above components, it was prepared so as to be 4 g per 1 ring according to a conventional method.

<1-5> Preparation of granules

0.15 mg of extract of the present invention

glucose 200 mg

Starch 600 mg

After mixing the above ingredients, 100 mg of 30% ethanol was added and dried at 60° C. to form granules, and then filled in a bag.

<Preparation Example 2> Preparation of health food containing the extract of the present invention as an active ingredient

<2-1> Manufacture of wheat flour food

0.5 to 5.0 parts by weight of the extract of the present invention was added to wheat flour, and breads, cakes, cookies, crackers and noodles were prepared using this mixture.

<2-2> Preparation of soups and gravies

By adding 0.1 to 5.0 parts by weight of the extract of the present invention to soup and broth, processed meat products for health promotion, soup and broth of noodles were prepared.

<2-3> Preparation of ground beef

Ground beef for health promotion was prepared by adding 10 parts by weight of the extract of the present invention to ground beef.

<2-4> Production of dairy products

5 to 10 parts by weight of the extract of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-5> Manufacture of wire type

Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder.

Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared into powder having a particle size of 60 mesh with a grinder.

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

The grains, seeds and extracts of the present invention prepared above were prepared by blending in the following ratios.

Grains (30 parts by weight of brown rice, 15 parts by weight of barley, 20 parts by weight of barley),

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame),

The extract of the present invention (3 parts by weight),

Reishi (0.5 parts by weight),

Rehmannia (0.5 parts by weight)

<Preparation Example 3> Preparation of a health drink containing the extract of the present invention as an active ingredient

<3-1> Manufacturing of health drinks

After instant sterilization by homogeneously mixing 100 mL of the extract of the present invention with auxiliary materials such as high fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), and water (75%) This was prepared by packaging in small packaging containers such as glass bottles and plastic bottles.

<3-2> Preparation of vegetable juice

Vegetable juice was prepared by adding 100 mL of the extract of the present invention to 1,000 mL of tomato or carrot juice.

<3-3> Preparation of fruit juice

Fruit juice was prepared by adding 100 mL of the extract of the present invention to 1,000 mL of apple or grape juice.

<Preparation Example 4> Preparation of an external preparation containing the extract of the present invention as an active ingredient

<4-1> Preparation of ointment

An ointment containing the extract of the present invention was prepared in a conventional manner according to the composition described in Table 1 below.

ingredient content(%) extract of the present invention 1.0 glycerin 8.0 butylene glycol 4.0 liquid paraffin 15.0 Ketaglucan 7.0 Carbomer 0.1 Caprylic/Capric Triglycerides 3.0 squalane 1.0 cetearyl glucoside 1.5 Sorbitan Stearate 0.4 cetearyl alcohol 1.0 beeswax 4.0 antiseptic a very small amount Spices a very small amount Purified water remaining amount Sum 100

<Preparation Example 5> Preparation of a cosmetic composition containing the extract of the present invention as an active ingredient

<5-1> Nutrient lotion

Nutrient lotion containing the extract of the present invention was prepared in a conventional manner with the component ratios shown in [Table 2] below.

ingredient content (wt%) extract of the present invention 2.0 Glyceryl Stearate SE 1.5 cetearyl alcohol 1.5 lanolin 1.5 Polysorbate 60 1.3 sorbitastearate 0.5 hydrogenated vegetable oil 4.0 mineral oil 5.0 trioctanoin 2.0 dimethicone 0.8 tocopherol acetate 0.5 carboxyvinyl polymer 0.12 glycerin 5.0 1,3-butylene glycol 3.0 Sodium Hyaluronate 5.0 triethanolamine 0.12 Uniside-You 13 0.02 incense a very small amount Distilled water remaining amount Sum 100

<5-2> Nourishing Cream

A nourishing cream containing the extract of the present invention was prepared by a conventional method with the component ratios shown in [Table 3] below.

ingredient content (wt%) extract of the present invention 2.0 lipophilic monostearate glycerin 1.5 cetearyl alcohol 1.5 stearic acid 1.0 Polysorbate 60 1.5 sorbitastearate 0.6 isostearyl isosterate 5.0 squalane 5.0 mineral oil 35.0 dimethicone 0.5 Hydroxyethyl Cellulose 0.12 glycerin 6.0 triethanolamine 0.7 Uniside-You 13 0.02 incense a very small amount Distilled water remaining amount Sum 100

Claims (16)

Evening primrose ( Oenothera biennis ) A pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria, containing an extract as an active ingredient.
The pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria according to claim 1, wherein the evening primrose extract is extracted with water, a C ₁ to C ₅ lower alcohol or a mixed solvent thereof.
The method according to claim 1, wherein the evening primrose extract is extracted with one or more solvents selected from the group consisting of water, methanol, ethanol, ethyl acetate, acetone, ether, chloroform, benzene, hexane, dichloromethane, and mixed solvents thereof. A pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria.
The method of claim 1, wherein the evening primrose extract comprises one or more extracts selected from the group consisting of flowers, leaves, branches, fruits, roots, stems and seeds of evening primrose. A therapeutic pharmaceutical composition.
The pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria according to claim 1, wherein the evening primrose extract is contained in a concentration of 10 μg/ml to 100 mg/ml.
The method according to claim 1, wherein the pathogenic bacteria is Escherichia coli O157:H7 or Staphylococcus aureus , and the non-pathogenic bacteria are Escherichia coli DH5α ( Escherichia coli DH5α ) or Propionibacterium acnes . A pharmaceutical composition for preventing or treating infections caused by pathogenic and non-pathogenic bacteria.
According to claim 1, wherein the infection caused by pathogenic and non-pathogenic bacteria is conjunctivitis, abscess, dermatitis, acne, enteritis, vaginitis, hemolytic uremic syndrome, hemolytic colitis, osteoarthritis, peritonitis, pneumonia, osteomyelitis, cellulitis, meningitis, nephritis, carditis , gastritis, esophagitis, duodenitis, colitis, bacteremia, pneumonia, toxin shock syndrome, food poisoning, high fever, sepsis, acute breast cancer, edema disease, mastitis, enterocoliosis, renal dysfunction, constipation, inflammatory bowel disease, dyspepsia, gastro reflux , Abdominal distension, gas, recovery discomfort, pain, diarrhea, irritable bowel syndrome and a pharmaceutical composition for the prevention or treatment of infections caused by pathogenic and non-pathogenic bacteria, characterized in that at least one selected from the group consisting of a feeling of indigestion.
Evening primrose ( Oenothera biennis ) A health functional food composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an extract as an active ingredient.
Evening primrose ( Oenothera biennis ) A food composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an extract as an active ingredient.
An external preparation for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, containing an evening primrose ( Oenothera biennis ) extract as an active ingredient.
A cosmetic composition for preventing or improving infection caused by pathogenic and non-pathogenic bacteria, comprising an evening primrose extract as an active ingredient.
A composition for inhibiting the growth of pathogenic and non-pathogenic bacteria, containing an evening primrose extract as an active ingredient.
The method according to claim 12, wherein the pathogenic bacteria is Escherichia coli O157:H7 or Staphylococcus aureus , and the non-pathogenic bacteria are Escherichia coli DH5α ( Escherichia coli DH5α ) or Propionibacterium acnes . A composition for inhibiting the growth of pathogenic and non-pathogenic bacteria.
13. The method of claim 12, wherein the composition for inhibiting the growth of pathogenic and non-pathogenic bacteria is selected from the group consisting of pharmaceutical compositions, health functional foods, livestock feed additives, industrial disinfectants, industrial preservatives, antibacterial daily products, antibacterial cosmetics, and antibacterial food or pharmaceutical additives. A composition for inhibiting the growth of pathogenic and non-pathogenic bacteria, characterized in that the use.
An antibacterial composition for pathogenic and non-pathogenic bacteria, containing an evening primrose extract as an active ingredient.
The method according to claim 15, wherein the pathogenic bacterium is Escherichia coli O157:H7 or Staphylococcus aureus , and the non-pathogenic bacterium is Escherichia coli DH5α ( Escherichia coli DH5α ) or Propionibacterium acnes . to, an antibacterial composition for pathogenic and non-pathogenic bacteria.

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