TWI477279B - The use of lactic acid bacteria belonging to the genus Enterococcus for the manufacture of a composition having a bed sores - Google Patents

Description

Use of a lactic acid bacterium belonging to the genus Enterococcus for the manufacture of a composition having an acne-promoting effect

The present invention relates to a composition having an acne-improving effect. More specifically, the present invention relates to a composition having an acne-promoting action containing lactic acid bacteria as an active ingredient, and use as a medicine, a reagent, and a food or drink thereof.

Acne is a state in which the blood flow to the skin, subcutaneous fat tissue, and muscles is interrupted due to continued oppression, resulting in the death of these tissues. In the prevention or treatment of acne, local treatment including care, prevention, nutrition management, and skin care for the purpose of dispersing body pressure is carried out to ensure local blood flow. In the local treatment of acne, it is necessary to correctly determine the state of acne, select a treatment suitable for this state, external medicine and wound coating materials, and apply to patients. In the previous topical treatment of acne, due to the need for a high degree of professional knowledge, it is required to develop a new and simpler treatment.

In addition, in recent years, the following attempts have been made to verify the preventive effects of diseases and the improvement of disease symptoms, such as lactic acid bacteria, and to use them as medicines that can be easily ingested and have high safety. Food and beverage.

For example, the lactic acid bacteria isolated from human intestinal tract, the Enterococcus faecalis EC-12 strain (hereinafter sometimes referred to as "EC-12"), the applicant reported that it has the effect of inhibiting the atrophy of the villi and the inhibitory effect of intestinal epithelial cell proliferation ( Patent Document 1, Non-Patent Document 1), improvement effect of intestinal flora, laxative promoting effect, and mechanism of laxative improvement (Patent Document 2, Non-Patent Documents 2 to 4), organisms infected with Listeria The defensive effect (Non-Patent Document 5), the elimination effect of the drug-resistant bacteria (Patent Document 3), the activation effect of natural immunity and acquired immunity (Non-Patent Document 6), and the inflammation-inhibiting effect on contact dermatitis (Patent Document 4) The alleviation effect of type I allergic reaction (asthma, pollinosis, atopic dermatitis) (Patent Document 5, Non-Patent Document 7), Rheumatoid Arthritis Improvement Effect (Patent Document 6), Han Sin (Leprosy) The preventive effect (Patent Document 7), the antitumor effect (Non-Patent Document 8), and the healing effect of burns (Non-Patent Document 9).

In addition to the above-mentioned report, it has been reported that intestinal bacteria such as lactic acid bacteria have been reported to improve the symptoms of various diseases, but there have been no reports of improvement in acne.

Prior technical literature Patent literature

Patent Document 1: Japanese Patent Laid-Open Publication No. 2008-255080

Patent Document 2: Japanese Patent Laid-Open Publication No. 2004-51530

Patent Document 3: Japanese Patent Laid-Open No. 2006-89421

Patent Document 4: Japanese Patent Laid-Open Publication No. 2006-28050

Patent Document 5: Japanese Patent Laid-Open Publication No. 2007-91694

Patent Document 6: Japanese Patent Laid-Open Publication No. 2007-254333

Patent Document 7: Japanese Patent Laid-Open Publication No. 2007-290992

Non-patent literature

Non-Patent Document 1: Yoshida Yoshiko and others, the 62nd Japan Nutrition and Food Society Conference Lecture Highlights p247 (2008)

Non-Patent Document 2: Microb. Ecol. Health Dis., 17(2), 107-113 (2005)

Non-Patent Document 3: Microb. Ecol. Health Dis., 16(4), 188-194 (2004)

Non-Patent Document 4: Biosci. Biotechnol. Biochem., 71, 60589-1-4 (2007)

Non-Patent Document 5: J. New Rem. & Clin., 53(3), 78-88 (2004)

Non-Patent Document 6: Animal Sci. J., 78, 92-97 (2007)

Non-Patent Document 7: Skin Research, Suppl. 10, 38-44 (2008)

Non-Patent Document 8: Otsuka Masahiro et al., The 15th Japanese Society of Biotherapeutics transcripts p110 (2002)

Non-Patent Document 9: Yamada Kaoru and others, the 147th Japan Veterinary Society Academic Conference Lecture Highlights p295 (2009)

The present invention has been made in view of the above circumstances, and an object thereof is to provide a composition which has an acne-improving action, can be easily administered or ingested, and has high safety.

In order to solve the above problems, the inventors of the present invention have studied the effect of lactic acid bacteria in a mouse model of acne. Specifically, this mouse model was ingested as Enterococcus faecalis (EC-12 strain) which is an example of a lactic acid bacterium, and its acne-improving effect was evaluated. As a result, in the mouse model, the reduction in the area of the acne was confirmed from the early stage after the formation of the acne, and the high expression of various inflammatory cytokines or growth factors was confirmed in the early stage after the formation of the acne. Promote wound healing. Further, it can be judged that the expression of inflammatory cytokines or growth factors is reduced in the latter stage after the formation of acne, and the normal state can be restored more quickly than the control. In this way, Enterococcus faecalis (EC-12 strain) showed an improvement from the early stage after the formation of acne. Further, based on the comparison between the negative control group and the group administered with Enterococcus faecalis (EC-12 strain) on the same day, it was also determined that the effect was to prevent the administration of Enterococcus faecalis (EC-12 strain) before the formation of hemorrhoids. This is especially true.

In view of the above, the inventors of the present invention have found that Enterococcus faecalis (EC-12 strain) is extremely useful as a medicine or a food or drink for the prevention of acne or the early stage of healing, thereby completing the present invention.

In more detail, the present invention provides the following inventors.

(1) A composition for improving acne, which comprises lactic acid bacteria as an active ingredient.

(2) The composition according to (1), wherein the lactic acid bacteria belong to the genus Enterococcus.

(3) The composition of (1), wherein the lactic acid bacterium is Enterococcus faecalis.

(4) A composition according to any one of (1) to (3) which is a pharmaceutical composition.

(5) A composition according to (4) which is administered orally.

(6) A composition according to (4) or (5) which is administered prophylactically prior to the formation of acne.

(7) The composition according to any one of (1) to (3) which is a food or drink.

(8) A composition according to (7) which is an oral ingestor.

(9) A composition according to (7) or (8) which is taken before acne formation.

(10) A composition according to any one of (4) to (9), which is additionally provided with a subject for improving the indication of acne.

The improvement of acne in a living body can be achieved by administering the composition of the present invention in the form of a pharmaceutical product or ingesting it in the form of a food or drink. In particular, a remarkable effect can be obtained by prophylactically administering the composition of the present invention before the formation of acne. The compositions of the invention are useful in the prevention of acne or in the early stages of acne. Further, the active ingredient of the composition of the present invention is lactic acid bacteria and has high safety. Therefore, it can be administered orally when used as a medicine, and unlike the previous topical treatment of acne, it is simple and has a small burden on acne patients. Moreover, the composition of the present invention is a food or drink such as a health food, and a healthy person can easily take it in daily life to prevent acne.

The present invention provides a composition for improving acne comprising lactic acid bacteria as an active ingredient.

In the present invention, the term "acne" refers to a state in which blood flow to the skin, subcutaneous fat tissue, and muscle is interrupted due to continued compression, thereby causing the tissues to die. In the present invention, the "improving action of acne" means that it is difficult to produce acne by the administration or ingestion of lactic acid bacteria, the acne produced is mild, the deterioration of acne is suppressed, and the healing of acne is accelerated. Also, in the process of forming the acne to cure, it includes not only improving the overall situation, but also improving the specific stage. As shown in the present embodiment, the improvement effect of acne can be evaluated, for example, by measurement of the area of acne or measurement of the expression of inflammatory cytokines or growth factors.

The "lactic acid bacteria" which is an active ingredient of the composition of the present invention can be exemplified by Enterococcus, Bifidobacterium, Lactobacillus, Streptococcus, and Lactococcus, but there is no particular effect as long as it has an acne-improving effect. limit.

Examples of the lactic acid bacteria belonging to the genus Enterococcus include Enterococcus faecalis and Enterococcus faecium, and bacteria belonging to the genus Bifidobacterium include, for example, Bifidobacterium breve, Bifidobacterium breve, and Bifidobacterium breve Examples of the bacterium belonging to the genus Lactobacillus, such as Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus saliva, and bacteria belonging to the genus Streptococcus, for example, Streptococcus thermophilus, and the like, are bacteria belonging to the genus Lactococcus. For example, Lactococcus lactis, Lactococcus lactis, etc. are mentioned. The lactic acid bacteria used in the composition of the present invention preferably belong to the genus Enterococcus, and particularly preferably Enterococcus faecalis.

Furthermore, the Enterococcus faecalis EC-12 strain (EC-12) used in the present example was deposited at the Patent Bio-Reservation Center of the Industrial Technology Research Institute of the Independent Administrative Corporation on February 25, 2005 (the original registration date). 305-5466, 6th, Central, 1st, 1st, 1st, Tsukuba, Ibaraki, Ibaraki, Japan (trustee number is FERM BP-10284). As Enterococcus faecalis, strains such as ATCC19433, ATCC14508, ATCC23655, IFO16803, and IFO16804 can be used in addition to the EC-12 strain.

These lactic acid bacteria may be used alone or in combination with two or more species in the composition of the present invention. Further, in the composition of the present invention, the lactic acid bacteria can be used in the form of live bacteria and in the form of dead cells. The dead cells are preferred as an active ingredient of the composition of the present invention in terms of excellent heat resistance, stable quality, and odorless and odorless. As the dead cells in the composition of the present invention, heat-sterilized cells can be used. The sterilized cells can be prepared by, for example, filtration, centrifugation, or the like, and recovering the cells from the culture obtained by culturing the lactic acid bacteria according to the usual method, washing them in water, and then suspending them in water or the like. After heating at 120 ° C or lower (preferably 80 to 120 ° C) for 30 minutes or less (3 seconds to 30 minutes), it is concentrated and dried as necessary. In addition, the fine powder of the heat-sterilized cells of the Enterococcus faecalis EC-12 strain is commercially available as the trade name "EC-12 (EF Power)" (manufactured by Combi Co., Ltd.).

The composition of the present invention may be in the form of a pharmaceutical composition having a acne-improving action, a food or drink (including an animal feed), or an agent used in an animal model test or the like.

In the present example, when the acne mouse model was ingested with Enterococcus faecalis (EC-12 strain), a significant reduction in the area of acne was confirmed from the early stage after the formation of acne. In addition, this effect is particularly large in the case where the Enterococcus faecalis (EC-12 strain) is administered prophylactically before the formation of acne. Therefore, the composition of the present invention can be preferably used as a prophylactically administered pharmaceutical composition or a preventive (daily) ingested food or drink, whereby the prevention of acne or the healing of an early stage can be performed.

Moreover, when the Enterococcus faecalis (EC-12 strain) was taken up in the mouse model of the present example, it was confirmed that inflammatory cytokines or growth factors were highly expressed in comparison with the control at the initial stage after the formation of acne. The wound healing was promoted; further, it was confirmed that the expression of inflammatory cytokines or growth factors was reduced in comparison with the control, and the normal state was restored faster than the control. Therefore, the composition of the present invention can also be used as an inflammatory cytokine or growth factor for promoting the initial stage of acne formation (for example, IL-1α gene, TNF-α gene, TGF-β gene, FGF-7 gene, FGF-10). A composition for inhibiting the expression of a gene, and an inflammatory cytokine or growth factor (for example, IL-1α gene, IL-8 gene, TNF-α gene, TGF-β gene, FGF-) for promoting post-acne formation. A composition inhibiting the expression of 10 genes).

The composition of the present invention can be formulated by a known pharmaceutical method. For example, capsules, troches, pills, liquids, powders, granules, fine granules, film coating agents, round granules, buccal agents, sublingual agents, chewables, buccal agents, ointments (paste agent), syrup, suspending agent, expectorant, emulsion, coating agent, ointment, plaster, paste, percutaneous absorption preparation, lotion, attractant, aerosol, injection, suppository, etc. Used orally or non-orally. The present invention is a composition comprising an enteric bacterium, that is, a lactic acid bacterium, as an active ingredient, which can be easily administered or ingested by oral administration. That is, a preferred method of administration or ingestion of the composition of the present invention is oral administration or ingestion.

In the formulation, it can be used as a carrier for pharmacological or food or beverage, specifically sterilized water or physiological saline, vegetable oil, solvent, main ingredient, emulsifier, suspending agent, surfactant, stabilizer. , flavoring agents, fragrances, excipients, vehicles, preservatives, binding agents, diluents, isotonic agents, analgesics, extenders, disintegrating agents, buffers, coating agents, lubricants, colorants Suitable combinations of sweeteners, thickeners, flavoring agents, dissolution aids or other additives.

When the composition of the present invention is used as a pharmaceutical composition, it may be used in combination with a known pharmaceutical composition for the prevention or treatment of acne.

When the composition of the present invention is used as a food or drink, the food or drink may be, for example, a health food, a functional food, a specific health food, a nutritional supplement, a patient food, or an animal feed. The food or drink of the present invention can be ingested in the form of a composition as described above, and can also be ingested in the form of various foods and drinks. Specific examples of foods and drinks include various beverages such as fruit and vegetable juices, refreshing beverage waters, tea beverages, beverages, jelly-like beverages, and functional beverages; alcoholic beverages such as beer; rice, noodles, breads, and doughs. Such as carbohydrate-containing foods; fish ham, sausages, aquatic products and other products; curry, pouring vegetables, Chinese soup and other cooking products; soups; milk, milk drinks, ice cream, cheese, yogurt and other dairy products; beans Sauce, yoghurt, lactic acid bacteria, lyophilized beverages, pickled products, etc.; soy products; western-style snacks such as biscuits and cookies; Japanese-style snacks such as taro or mutton; confectionery, chewing gum, jelly, jelly, pudding Various kinds of snacks such as frozen desserts or frozen desserts; fast-food soups such as instant soups and instant soups, and microwave ovens for foods. Further, a healthy food or drink prepared in the form of a powder, granules, a tablet, a capsule, a liquid, a paste or a jelly may also be mentioned.

The composition of the present invention can be used as an object for animals including humans, and is not particularly limited as an animal other than human, and can be used for various livestock, poultry, pets, laboratory animals, and the like. Specific examples include pigs, cows, horses, sheep, goats, chickens, wild ducks, ostriches, ducks, dogs, cats, rabbits, hamsters, mice, rats, monkeys, etc., but are not subject to such restrictions. .

The manufacture of the food or beverage of the present invention can be carried out using manufacturing techniques well known in the art. One or two or more components (for example, nutrients) which are effective for the improvement of acne can be added to the food or drink. Further, it is also possible to produce a multifunctional food or drink by combining with other components or other functional foods that function in addition to the improvement of acne.

In the case of administering or ingesting the composition of the present invention, the dose or intake may be based on the age, body weight, symptoms of acne, state of health, type of composition (medicine, food, etc.), etc. Appropriate choice. For example, the dosage or intake of the lactic acid bacteria is usually from 0.01 mg/kg body weight to 100 mg/kg body weight, preferably from 1 mg/kg body weight to 10 mg/kg body weight. As such, the present invention also provides a method for improving acne in a subject, which is characterized in that a lactic acid bacterium is administered to a subject or a lactic acid bacterium is ingested by a subject.

The article (medicine, food or drink, reagent) of the composition of the present invention or a specification thereof may be an additional subject matter for the purpose of improving acne. The term "additional labeling in the product or the specification" as used herein refers to the labeling of the body, container, packaging, etc. of the product, or the labeling of the product information, attached documents, promotional materials, other printed materials, and the like. The subject matter of the indication for improving acne may include information on the mechanism by which acne is improved by administering or ingesting lactic acid bacteria. As the mechanism, for example, information on inflammatory cytokines or growth factors associated with acne (for example, IL-α gene, IL-8 gene, TNF-α gene, TGF-β gene, FGF-7 gene, etc.) may be cited. The role or performance of the FGF-10 gene).

Example

Hereinafter, the present invention will be more specifically described based on examples and comparative examples, but the present invention is not limited to the following examples.

[Example 1] Study on the improvement effect of EC-12 on acne in a mouse model

A BALB/c 10-week old male mouse was used as a test animal. "Normal saline administration group (control group)", "preventive EC-12 administration group", and "the same day EC-12 administration group" were set as test groups, and 15 (45 mice) mice were used for each group. .

After acclimation of the mouse, the hair was shaved under the anesthesia using a commercially available hair clipper, and the hair removal cream was used to remove the hair. Carefully wipe the hair removal part with distilled water. On the next day of shaving, lift the center of the back and hold it for two hours with two magnet plates (diameter 12 mm × thickness 5 mm, 1180 G) and then remove the magnet for 12 hours (9 o'clock) Installation, 21 o'clock removal). This cycle was repeated for 3 consecutive days to make acne. Furthermore, individual feeding is performed only when making acne. When making acne, butyl morphine (2 mg/kg) was administered subcutaneously as an analgesic. The method for producing acne is carried out in accordance with Stadker et al. (J. Invest. Surg. 2004. 17:221-227).

In the "preventive EC-12 group" and "the same day EC-12 group", EC-12 was suspended in normal saline, and EC-12 became 10 mg/kg (body weight) daily. Mice were forced to orally administered. For the "salt-administered group", sterilized physiological saline is orally administered. The "preventive EC-12 group" is administered with EC-12, and the "saline group" is administered to the "salt group". The saline system is started from 7 days before the start of the acne. Check it out. The "EC-12 group on the same day" was administered to the EC-12 line from the acne production day, once a day at 9 am until the necropsy.

For the state of the wound, the photograph was taken daily and photographed, and the area of the wound was measured by image analysis of the photograph. After making acne, 5 of each group were examined on the 3rd, 6th, and 11th day. Schering-Plough was administered intraperitoneally, and after deep anesthesia, it was slaughtered by bloodletting. The wound area reduction rate was calculated as the value when the maximum acne area of the "preventive EC-12 administration group" was set to 100.

As a result, regarding the reduction rate of the wound area, the "preventive EC-12 administration group" performance was compared with the "EC-12 injection group on the same day" and the "salt saline administration group" on the fourth day after the formation of the acne. A lower value (Figure 1).

The inventors of the present invention further confirmed the mRNA expression of cytokines and growth factors.

Among the known cytokines, IL-8 is stimulated by inflammatory cytokines such as IL-1 or TNF, and is composed of white blood cells represented by white blood cells, and fibroblasts or I skin cells. Factor (Leukocyte Chemotactic Factor). Further, IL-1α has various physiological activities in various immunocompetent cells, but as its most important role, it is known that IL-2 production can be induced by helper T cells, and T cells are promoted via IL-2. Differentiation and proliferation. Further, TNF-α causes fever or various inflammatory reactions via production of IL-1, PGE2, collagenase, etc., and thus it is considered to be one of the vehicles in the inflammatory reaction.

Further, among the known growth factors, TGF-β promotes the chemotaxis of macrophages to the injured site when the tissue is damaged. Since the macrophage itself secretes TGF-β, the secretion can increase the fibroblasts, thereby synergistically promoting the wound healing process. Further, FGF-7 is a multifunctional signal molecule which exhibits activity such as proliferation and differentiation for fibroblasts and various cells, and is called KGF (Keratinocyte Growth Factor). Further, FGF-10 belongs to the subfamily of FGF-7 in the fibroblast proliferation factor family.

In the present example, the expression of the above-mentioned IL-1α, IL-8 and TNF-α as cytokines, and the above-mentioned TGF-β, FGF-7 and FGF-10 as growth factors were examined.

Specifically, after extracting RNA from the wound sample, cDNA was synthesized by reverse transcription, and the primer was used as a template, and the primer described in Table 1 was used for real-time PCR (Polymerase Chain Reaction) (Rotor-Gene 6200). (Qiagen))).

Further, a QuickGene 810 system and a QuickGene RNA tissue kit SII (Fuji Film Co., Ltd.) were used for RNA extraction, and a PrimeScript RT reagent kit (Perfect Real Time; Takara Bio) was used for reverse transcription. The operation is carried out in accordance with the operating instructions. The measured value was corrected by GAPDH (glyceraldehyde-3-phosphate dehydrogenase).

As a result, the amount of mRNA expression in the skin acne tissue of the "prophylactic EC-12 administration group" was compared with the inflammatory cytokines other than IL-8 and all growth factors on the third day after the production of acne. It was raised on the same day, the EC-12 was administered to the group and the "salt was administered to the group" (Table 2, Table 3). On the other hand, on the 6th day after the production of acne, in the "prophylactic EC-12 administration group", all inflammatory cytokines and growth factors other than FGF-7 were compared with "the same day EC-12 administration group". And "normal saline administration group" was reduced (Table 2, Table 3).

The process from the generation of hemorrhoids to the cure is usually divided into four stages: (1) bleeding clotting period, (2) inflammatory phase, (3) proliferative phase, and (4) maturity, compared to the same day EC-12 administration. In the "preventive EC-12 administration group", a significant reduction in the area of acne was confirmed on the fourth day of acne production. Moreover, the high expression of inflammatory cytokines and growth factors was observed on the third day of acne production, and thereafter, on the sixth day of acne production, the performance was drastically lowered. Based on these facts, it has been revealed that the administration of EC-12, especially since the formation of acne, can have an effect on the healing of acne at the beginning of acne. Also, based on these facts, it is suggested that EC-12 can also be used for the prevention of acne.

Industrial availability

As described above, the composition of the present invention can be administered in the form of a pharmaceutical product or ingested in the form of a food or drink, thereby improving the acne in the living body. The lactic acid bacteria which are active ingredients of the composition of the present invention are enteric bacteria which can be administered or ingested orally. Since the composition of the present invention has high safety and can be easily administered or ingested, it can be expected to be used as a new medicine or health food having an effect of improving acne.

<110> Rishang Kangbei Co., Ltd.

<120> Composition with acne improving effect

<130> P09-020

<140> 100106584

<141> 2011-02-25

<150> 2010-042303

<151> 2010-02-26

<160> 12

<170> PatentIn version 3.1

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Figure 1 shows the reduction rate of acne area from the 3rd to the 11th day after the formation of acne in the "preventive EC-12 administration group", "the same day EC-12 administration group", and "saline administration group". Figure. The maximum acne area of the "preventive EC-12 administration group" was set to 100, and the wound area reduction rate was calculated.

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Claims (4)

A use of a lactic acid bacterium belonging to the genus Enterococcus, which is used in the manufacture of a pharmaceutical composition for the improvement of acne. The use of claim 1, wherein the lactic acid bacteria is Enterococcus faecalis. The use of claim 1, wherein the pharmaceutical composition is an oral donor. The use of any one of claims 1 to 3, wherein the pharmaceutical composition is administered prophylactically prior to the formation of acne.