WO2019221555A1 - Composition comprising aster koraiensis extract or fraction thereof for wound treatment or skin regeneration - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising an Aster koraiensis extract or a fraction thereof for wound prevention or treatment, and to a quasi-drug product composition or cosmetic composition comprising an Aster koraiensis extract or a fraction thereof for skin regeneration. The Aster koraiensis extract or fraction thereof of the present invention can accelerate wound healing and skin surface regeneration. Therefore, the Aster koraiensis extract or fraction thereof of the present invention would be able to be widely utilized in the development of a pharmaceutical composition for wound prevention or treatment.

Description

A composition for wound healing or skin regeneration comprising bee odor extract or fractions thereof

The present invention relates to a pharmaceutical composition for the prevention or treatment of wounds, including a bee-flavored extract or fractions thereof, and to a quasi-drug composition or cosmetic composition for skin regeneration comprising a bee-flavored extract or fractions thereof.

Wound may be defined as the integrity of the skin epithelial tissue is destroyed by external pressure or the like. The normal wound healing process involves three stages: the inflammatory response, the formation of granulation tissue, the formation of new epithelial cells and new blood vessels. When these mechanisms are in place, complete healing is possible. However, when the wound recovery process is abnormally developed, the wound recovery may be delayed, and the angiogenesis or granulation tissue formation may be significantly slower than that of normal people due to the disorder caused by a specific disease.

Growth factor offers the possibility of treating malignant or chronic wound wounds by inducing the activity of various cells. Growth factors regulate many factors necessary for cell activity, such as cell division and migration and angiogenesis. Growth factors related to this process include EGF (epidermal growth factor), PDGF (platelet derived growth factor), FGF (fibroblast growth factor), KGF (keratinocyte growth factor), TGF-β (transforming growth factor-β), G-CSF (granulocyte colony stimulating factor), VEGF (vascular endothelial growth factor), and the like.

Normal wound wound recovery can be achieved through a combination of sequential processes such as cell migration and proliferation and induction of extracellular matrix (ECM) deposition. If a problem occurs in some of the above processes, the pathological inflammatory response may continue, leading to chronic bruising. Common causes of chronic ulcers include ischemia, diabetes, venous congestion, and unnecessary pressure on the wound. One or more causes may be active at the same time.

In the wound healing process, various factors such as inflammatory mediators, growth factors, and cytokines work in combination, and although an inflammatory response is necessary for wound healing, excessive inflammatory response can delay wound healing. Therefore, when regulating the expression of various factors acting on the inflammatory response, the wound healing process may be more smoothly performed.

On the other hand, Aster koraiensis is a perennial herb belonging to the Asteraceae family , which grows in well-lit and wet swamps, and is a Korean specialty plant growing in the south of Gyeonggi-do and Jeju-do. It is often used for horticulture or dermis, and it is sometimes used as a herb for boiled soft leaves or as a herb, or as a root for sputum and asthma.

Accordingly, the present inventors have made intensive research efforts to develop an excellent natural product-derived wound treatment, and confirmed that the bee-flavored extract or its fractions can cure wounds through ulcerative wound treatment and wound regeneration of the skin epithelium in a wound-treatment animal model. The present invention has been completed.

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of wounds comprising a bee odor extract or fractions thereof.

It is another object of the present invention to provide a method for preventing or treating wounds, comprising administering a bee anesthetist extract or a fraction thereof to an individual except a human.

Yet another object of the present invention is to provide a quasi-drug composition for skin regeneration comprising a bee odorous extract or a fraction thereof.

Still another object of the present invention is to provide a cosmetic composition for skin regeneration comprising a bee odorous extract or a fraction thereof.

As the bee odor extract and fractions thereof of the present invention can promote the healing of wounds and promote the regeneration of the skin epidermis. It may be widely used in the development of pharmaceutical compositions for the prevention or treatment of wounds.

1 is a graph showing the results of evaluation of cell mobility of keratinocytes.

Figure 2 is a graph comparing the effect of wound treatment over time in an ulcer animal model.

3 is a graph comparing skin epidermal tissue thickness in ulcer animal models.

4 is a graph comparing the expression and activity of MMP-2 / 9 in skin tissue and keratinocytes.

One aspect of the present invention for achieving the above object, provides a pharmaceutical composition for the prevention or treatment of wounds comprising a bee odor extract or fractions thereof.

In addition, the present invention provides a use for the prevention or treatment of wounds of a composition comprising a bee herb extract or a fraction thereof.

As used herein, the term " Aster koraiensis " is a perennial herb belonging to the Asteraceae family , which grows in well-lighted and damp wetlands, and is a Korean specialty plant that grows in South Korea's Gyeonggi-do and Jeju-do. It is often used for horticulture or dermis, and it is sometimes used as a herb for boiled soft leaves or as a herb, or as a root for sputum and asthma.

The term "extract" in the present invention may refer to a liquid component obtained by immersing in a solvent and then extracted for a predetermined time at room temperature or warmed state, a solid content obtained by removing the solvent from the liquid component.

The hummingbird extract with prophylactic and therapeutic activity of wounds can be extracted from various organs of natural, hybrid, and mutant plants, and for example, plant tissue culture as well as roots, stems, leaves, flowers, trunks of fruits, peels of berries. Extractable from water.

In the present invention, the bee stinger extract may be extracted from the root, stem, leaf, flower, or fruit of the bee sting odor, it is not particularly limited thereto.

In the present invention, the bee odorous extract is extracted with a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms and mixed solvents thereof, but is not limited thereto. In the present invention, the extract may include an extract obtained by an extraction treatment, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or any one of these modifiers or purified products.

The alcohol may be butyl alcohol, ethyl alcohol, methyl alcohol or a water soluble alcohol thereof. Specifically, the water-soluble alcohol may be one that contains 0.1 to 100% by weight alcohol.

The term "fraction" in the present invention means the result obtained by performing the fractionation to separate a specific component or a specific component group from a mixture comprising various various components.

In the present invention, the fractionation method for obtaining the fraction is not particularly limited, and may be performed according to a method commonly used in the art. As a non-limiting example of the fractionation method, a method of obtaining a fraction from the extract by treating a predetermined solvent with an extract obtained by extracting a bee-flavored extract.

In the present invention, the fraction may be a hexane fraction, ethyl acetate fraction, butanol fraction or water fraction of the honey bee extract. The fractions may each comprise 0.01 to 100% by weight, more specifically 1 to 80% by weight relative to the total weight of the pharmaceutical composition.

The kind of the fractionation solvent used to obtain the fraction in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvents include polar solvents such as water and alcohols; And nonpolar solvents such as hexane, ethyl acetate, chloroform, dichloromethane, and the like. These may be used alone or in combination of two or more thereof. In the case of using the alcohol in the fractionation solvent, specifically, C ₁ to C ₄ alcohol may be used.

The term "wound" in the present invention may be defined as the integrity of skin epithelial tissue is destroyed by external pressure or the like. Types of wounds include abrasions. bruise. There are lacerations, cuts by blades, ulcers, ulcers, ulcers, gross ulcers, ulcers, etc., and in the form of wounds, epidermal peeling ducts, subcutaneous bleeding, sputum, acne, Reimbursement;

The term "prevention" in the present invention refers to any action that inhibits or delays the symptoms of a wound by administration of a composition comprising a bee-flavored extract according to the present invention or a fraction thereof.

The term "treatment" in the present invention refers to any action in which the symptoms of the wound are improved or beneficially changed by administration of a composition comprising a bee-flavored extract according to the present invention or a fraction thereof.

In one embodiment of the present invention was administered to the ulcerative wound animal model by administering the hummingbird extract, promoting the therapeutic effect of the wound (Fig. 2).

Therefore, through the above results, it was found that the bee-flavored extract has an effect on the wound treatment.

As used herein, the term "pharmaceutical composition" means that is prepared for the purpose of preventing or treating a disease, and can be used by formulating in various forms according to each conventional method. For example, it may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, and the like, and may be used in the form of external preparations, suppositories, and sterile injectable solutions. Specifically, it may be used in a form suitable for eye drop administration, for example, eye drops, creams, ointments, gels or lotions.

The composition of the present invention may be prepared in the form of a pharmaceutical composition for preventing or treating wounds, further comprising a suitable carrier, excipient or diluent commonly used in the manufacture of a pharmaceutical composition, wherein the carrier is an unnatural carrier ( non-naturally occuring carriers).

In the present invention, carriers, excipients and diluents that can be included in the pharmaceutical composition 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, and the like in the above extracts and fractions thereof. , Sucrose or lactose, gelatin and the like are mixed and prepared. 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 oils such as olive oil, injectable esters 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.

The dosage of the pharmaceutical composition of the present invention can be determined by those skilled in the art in consideration of the purpose of use, the degree of addiction of the disease, the age, weight, sex, history, or type of substance used as an active ingredient of the patient. For example, the pharmaceutical composition of the present invention may be administered at about 0.1 ng to about 1,000 mg / kg, specifically 1 ng to about 1,000 mg / kg, per adult, and the frequency of administration of the composition of the present invention is particularly limited thereto. However, it may be administered once a day or several times in divided doses. The dosage does not limit the scope of the invention in any aspect.

Another aspect of the invention provides a method for preventing or treating wounds comprising administering to the individual a bee-smelling extract.

In the present invention, the term "beetle odor", "extract", "wound" is as described above.

As used herein, the term "individual" may include, without limitation, mammals including wounded animals and the like, and specifically may include humans.

The hummingbird extract may be administered in single or multiple doses in a pharmaceutically effective amount.

The route of administration according to the wound prevention or treatment method of the present invention may be administered via any general route as long as it can reach the target tissue.

The bee anesthetist extract of the present invention is not particularly limited, but as desired, eye drops, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, It may be administered via a route such as rectal administration.

Another aspect of the present invention provides a quasi-drug composition for skin regeneration comprising a bee odor extract.

In the present invention, the term "beetle odor" and "extract" are as described above.

The term "skin regeneration" in the present invention means to restore the damaged area of the skin tissue again. The hummingbird extract may be to promote the regeneration of the skin when administered to the skin tissue.

In one embodiment of the present invention, as a result of administering the bee anesthetic extract to the damaged epidermal tissue of the skin, it was confirmed that the thickness of the regenerated epidermis is about twice as thick as the control, to confirm the skin regeneration effect of the bee anesthetic extract (Fig. 3).

As used herein, the term "quasi drug" refers to articles that have a lesser action than drugs among articles used for the purpose of diagnosing, treating, ameliorating, alleviating, treating, or preventing a disease of a human or animal. For example, quasi-drug products under the Pharmaceutical Affairs Law exclude products used for the purpose of medicines, and include products used for the treatment or prevention of diseases of humans / animals, and products with slight or no direct action on the human body.

Specifically, the quasi-drug may include external skin preparations and personal hygiene products. More specifically, it may be, but is not limited to, a disinfectant cleaner, a shower foam, a gagreen, a wet tissue, a detergent soap, a hand wash, or an ointment.

When the composition according to the present invention is used as an quasi-drug additive, the composition may be added as it is or used with other quasi-drugs or quasi-drug components, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the intended use.

Yet another aspect of the present invention provides a cosmetic composition for skin regeneration comprising a bee-flavored extract.

In the present invention, the term "beetle odor", "extract", "skin regeneration" is as described above.

In the present invention, the cosmetic composition is a solution, external ointment, cream, foam, nourishing lotion, flexible lotion, pack, flexible water, latex, makeup base, essence, soap, liquid cleaning agent, bath, sunscreen cream, sun oil, It may be prepared in a formulation selected from the group consisting of suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays, but It is not limited.

The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers formulated in general skin cosmetics, and as conventional components, for example, oil, water, surfactants, moisturizers, lower alcohols, thickeners , Chelating agents, pigments, preservatives, fragrances and the like may be appropriately blended, but is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide and the like. May be used, but is not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used, and especially in the case of a spray, additionally chlorofluorohydro Propellants such as carbon, propane / butane or dimethyl ether may be included, but are not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsion may be used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butylglycol oil, and the like can be used, and in particular, cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester May be used, but is not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, micro Crystalline cellulose, aluminum metahydroxyde, bentonite, agar or tracant may be used, but is not limited thereto. These may be used alone or in combination of two or more thereof.

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters may be used, but are not limited thereto. These may be used alone or in combination of two or more thereof.

In addition to the said essential component, you may mix | blend with the cosmetic composition of this invention the other component normally mix | blended with cosmetics as needed. Specifically, as a blending component which may be added, an oil-fat component, a humectant, an emollient, surfactant, organic and inorganic pigment, organic powder, a ultraviolet absorber, a preservative, a bactericide, antioxidant, a plant extract, a pH adjuster, alcohol, a pigment, Fragrances, blood circulation accelerators, cooling agents, restriction agents, purified water, and the like, but are not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1. Preparation of Beetle Anesthetic Extract

After cutting the leaves and stems of the hummingbird odor, the mixture was extracted at room temperature three times for three days each with 80% (v / v) ethanol, which is 10 times the weight. The extract was filtered, concentrated under reduced pressure, dried and lyophilized to obtain a bee-flavored alcohol extract (KIOM83A).

Example 2. Preparation of Wound Care Animal Model

The experimental animals were used for the experiment after 6 weeks of male Sprague Dawley (SD) rat (Orient Bio, Sungnam, Korea) without skin lesions. To induce diabetes, 70 mg / kg of streptozotocin (Sigma, MO, USA) dissolved in 0.1 M citric acid buffer (pH 4.5) was injected intraperitoneally. After one week, blood glucose was measured in a fasted state, and only the individuals with fasting glucose of 250 mg / dl or more were selected and the average blood sugar per group was evenly distributed. Each test group was 1) normal wound group (denoted by Nor), 2) diabetic wound group (denoted by DM), and 3) diabetic-causing group of 100 mg / kg bee odor extract (denoted by KIOM-83A). Divided into groups. Three weeks after the onset of diabetes, 10 mg / kg zolazepam (Zoletil, Virbac, Carros, France) and 10 mg / kg xylazine hydrochloride (Rumpun, Bayer, Frankfurt, Germany) were anesthetized and anesthetized, followed by anesthesia. The blade was removed using a razor blade and then disinfected with 70% alcohol and a full-thickness excision ulcer wound was made using a 6 mm punch (Biopsy Punzh, Ziefel, Germany). Beetle odor extract was orally administered once daily for 18 days from the day of the ulcer wound.

Experimental Example 1. Evaluation of Cell Mobility of Beetle Anesthesia Extract

Cell mobility was evaluated to confirm the effect of wound treatment at the cellular level. Culture-Insert 2 Well was removed after incubating HaCaT cells, keratinocytes, for 2 days in μ-Dish with Culture-Insert 2 Well. The experiment was performed after washing once with FBS-free medium. Each test group consisted of 1) a normal group (denoted CTL), 2) a honeybee extract (marked as KIOM-83A) 3 μg / ml treated group, 3) a 10mm honeycomb extract 10 μg / ml treated group, and 4) a 30 μg honeybee extract extracted. / ml treatment group was divided into a total of four groups. Thereafter, the cells were incubated for 8 hours in a CO ₂ incubator.

In order to confirm the effect on the cell mobility of the bee-mied extract, culture-Insert 2 Well was taken immediately after removal and photographed through a microscope until 8 hours to measure the area. Cell mobility assessment calculated percent migration over 8 hours.

As a result, as shown in Figure 1, in the normal group, about 14% of the cells migrated for 8 hours, and in the group treated with 30 μg / ml of the hummingbird anesthetic extract, about 19% of the cells migrated, the significant difference between the two groups I could confirm the difference.

Experimental Example 2. Analysis of wound healing efficacy

In order to confirm the wound healing efficacy of the bee-flavored extract, photographs were taken every day from immediately after the wound was wounded until the end of the drug administration.The size of the wound observed by the naked eye was measured, divided by the initial wound size, and converted into a percentage to determine the wound healing rate. Calculated.

As a result, as shown in FIG. 2, the wounds were treated in normal rats on average 11 days, and in the diabetic wound group induced after 18 days of hyperglycemia, the wounds were all healed for about 17 days. On the other hand, in the group administered bee anesthetic extract, the duration of wound treatment was 12 days, which was 4 days shorter than that of the diabetic wound group.

Experimental Example 3. Analysis of skin epithelial regeneration efficacy of honey bee extract

In order to confirm the skin epithelial regeneration efficacy of the bee-flavored extract, 18 days after drug administration, the autopsy was performed, the skin of the wound area was removed to a size of 1.5 × 1.5 cm, fixed in 10% neutral buffered formalin, and paraffin embedded in the skin tissue. I made a slide. Hematoxylin-eosin (H & E) staining was observed under light microscopy to evaluate the thickness of the new epidermis at the site of wound healing.

As a result, as shown in Fig. 3, the diabetic wound group did not have perfect regeneration of the epithelium as compared with the normal wound group, and thus the thickness of the regenerated skin epithelium was less than half of that of the normal wound group. The epithelium was found to be more than twice as thick as the diabetic wound group.

Therefore, it was possible to confirm the skin regeneration promoting effect of the bee anesthetic extract.

Experimental Example 4. Expression and Activity Analysis of Skin Tissue and Keratinocytes MMP-2 / 9

In order to confirm the effect on the expression of MMP-2 / 9 of bee-flavored extract, MMP-2 / 9 density was analyzed in skin sections.

Gelatin zymography was performed on keratinocytes in order to confirm the effect on the MMP-2 / 9 activity of the hummingbird extract.

As a result, as shown in FIG. 4, the expression of MMP-2 / 9 was increased in the diabetic wound group compared to the normal wound group, and the expression of MMP-2 / 9 was decreased in the normal wound group compared to the diabetic wound group. MMP-2 / 9 expression density was significantly increased in diabetic wound group compared with normal group, and MMP-2 / 9 expression level was decreased in honeycomb extract group.

In addition, quantitative levels of active MMP-2 / 9 in hyperglycemic conditions were significantly increased than in normal conditions. Beetle anesthetic extract reduced the increase of hyperglycemia-induced MMP-2 / 9 activity.

Therefore, it was confirmed that the destruction of the skin tissue layer was reduced through the inhibition of MMP-2 / 9 in hyperglycemia.

In the present specification, those skilled in the art of the present invention can fully recognize and infer the details that have been omitted, and the technical spirit or essential configuration of the present invention in addition to the specific examples described in this specification are changed. Many more variations are possible without departing from the scope of the invention. Therefore, the present invention can be implemented in a manner different from that specifically described and illustrated herein, which can be understood by those skilled in the art.

Claims (6)

1. Pharmaceutical composition for the prevention or treatment of wounds, including bee odor extract or fractions thereof.
2. The method of claim 1,

   The extract is a pharmaceutical composition that is extracted with a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms and mixed solvents thereof.
3. The method of claim 1,

   Wherein the fraction is an n-hexane fraction, an ethyl acetate fraction, a butanol fraction or a water fraction of the extract.
4. A quasi-drug composition for skin regeneration comprising a bee odor extract or a fraction thereof.
5. Cosmetic composition for skin regeneration comprising a bee odor extract or fractions thereof.
6. A method of preventing or treating wounds comprising administering a bee scabbard extract or a fraction thereof to a subject.

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