KR102112774B1 - Composition for preventing or treating acne comprising ethyl acetate fraction of sargassum miyabei yendo - Google Patents

Abstract

The present invention relates to a composition for preventing or treating acne containing the miabemozaban ethyl acetate fraction as an active ingredient, wherein the miabemozaban ethyl acetate fraction penetrates neutrophils induced by acne-causing bacteria Propionibacterium acnes As it is confirmed to exhibit the effect of inhibiting and reducing the abscess, the composition containing the miabemozaban ethyl acetate fraction as an active ingredient may be provided as an excellent acne treatment agent and a cosmetic composition for preventing or improving acne.

Description

Composition for preventing or treating acne comprising ethyl acetate fraction of sargassum miyabei yendo}

The present invention relates to a pharmaceutical composition and a cosmetic composition for the prevention or treatment of acne containing the miabemozaban ethyl acetate fraction as an active ingredient.

The causes of acne are not yet clearly defined, but if genetic and environmental factors are excluded, it is reported that three major causes are directly involved in acne development. The three main causes are briefly explained through the mechanism of acne.

The first cause can be said to be due to excess sebum. The formation of sebum is caused by the action of androgenic hormone, and the sebaceous gland function is enhanced through abnormal increase of the male hormone or several related mechanisms, resulting in excess sebum.

The second is explained by the closure of the pores due to hyperkeratinization of the hair follicles. The generated sebum should be smoothly discharged out of the skin, but due to hyperkeratization of the hair follicles, the pores are blocked and smooth discharge of the sebum is suppressed. In connection with sebaceous secretion, there are all the enzymes required to convert inactive steroid precursors into active androgens in the skin, 5α-reductase is one of them. Testosterone, one of the male hormones, is converted to dihydrotestosterone by 5α-reductase present in the cell after entering the sebaceous gland, and this signal is used to signal multi-step pathways such as gene expression. This results in sebum production.

The third cause is acne lesions due to dermatophytes such as Staphylococcus epidermis, Staphylococcus aureus and Propionibacterium acnes, which cause acne. It is explained as an important cause of exacerbation. Acne bacteria residing within the hair follicles form pore wall irritants. The micro-composite gradually increases in size as the sebum continues to secrete and develops into a compost. As a result, the hair follicle wall becomes thinner, and the irritating substances produced by acne bacteria stimulate the thinned hair follicle wall to gradually exacerbate the compost into inflammatory lesions. (Gujin → pustules → nodules → scars). On the other hand, various researches on methods for the treatment or improvement of acne have been conducted, focusing on sebum suppression and antibacterial effects on skin flora, depending on the cause of such acne.

Acne treatment drugs that have been known so far can be useful drugs for acne treatment by controlling sebum secretion, correcting keratinization, and inhibiting bacteria represented by propionium bacterium acnes. Specifically, the use of female hormones, such as anti-androgens to control the secretion of abnormal male hormones to suppress sebaceous hypersecretion, the use of antibacterial and antibiotic agents to suppress the growth of acne bacteria, non-steroidal system with anti-inflammatory action A method of using an anti-inflammatory agent is used, and in the cosmetic field, a method of suppressing skin exfoliation and inhibiting acne bacteria has been mainly used by adding ingredients such as lesocinol, sulfur, salicylic acid, and benzoyl peroxide.

However, if these ingredients are added excessively, skin irritation and easily dry side effects occur, so the compounding limit is set in the case of lesocinol and salicylic acid, and components such as benzoyl peroxide are designated as prohibited ingredients in Korea. .

Republic of Korea Patent Publication No. 10-2004-0056080 (released on June 30, 2004)

The present invention is to prevent acne induced by acne-causing bacteria and to provide a composition for preventing or treating acne containing natural miabemozaban with high skin stability as an active ingredient while exhibiting excellent therapeutic effect.

The present invention provides a pharmaceutical composition for preventing or treating acne containing the miabemozaban ethyl acetate fraction as an active ingredient.

In addition, the present invention provides a cosmetic composition for preventing or improving acne containing the miabemozaban ethyl acetate fraction as an active ingredient.

According to the present invention, the miabemozaban ethyl acetate fraction is confirmed to exhibit the effect of inhibiting the infiltration of neutrophils induced by acne-causing bacteria Propionibacterium acnes and reducing abscess, the miabemozaban ethyl acetate The composition containing the fraction as an active ingredient may be provided as an excellent acne treatment and cosmetic composition for preventing or improving acne.

1 is a schematic diagram showing the lesion site in which the acne bacterium Propionibacterium acnes was injected, 1 is a saline injection site, and 2 is a miabemosaban after acne bacteria (PA) injection The site where the ointment (vechile) not containing the ethyl acetate fraction was applied, and number 3 was the site where the ointment containing 50 μg of the miabemozaban ethyl acetate fraction (SMYEE) was applied after acne bacteria (PA) injection (PA: P. acnes ).
Figure 2 is an acne bacterium, Propionibacterium acnes (Propionibacterium acnes) in the lesion site is injected with an ointment containing 50 μg of miabemozaban ethyl acetate fraction (SMYEE) or ointment containing ointment, hemotox after confirming the change in abscess formation This is the result of staining of cylin & eosin.
Figure 3 is an acne bacterium Propionibacterium acnes (Propionibacterium acnes) to the lesion site is injected with the mibemozaban ethyl acetate fraction (SMYEE) ointment containing 50μg or ointment after application of the ointment confirmed the effect of suppressing neutrophil infiltration Histochemical staining results (PA: P. acnes, NIMP-R14: neutrophil marker).

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

Sargassum miyabei yendo is a brown algae belonging to the genus Sargassaceae and sargassum, and is mainly found in the South Coast and Jeju Island in Korea. Accordingly, the present inventors completed the present invention, while studying the physiological activity of miabemozaban, confirming that miabemozaban exhibits an excellent effect on acne treatment induced by acne bacteria.

The present invention can provide a pharmaceutical composition for preventing or treating acne containing the miabemozaban ethyl acetate fraction as an active ingredient.

The fraction may be a miabemozaban ethyl acetate fraction obtained by fractionating a miabemozaban ethanol extract with ethyl acetate, more preferably an ethyl acetate fraction of a miabemozaban extract extracted with a 70% aqueous ethanol solution.

The miabemozaban ethyl acetate fraction may inhibit abscess formation or neutrophil penetration induced by acne bacteria.

The acne can be induced by acne-causing dermatophytes consisting of Staphylococcus epidermis, Staphylococcus aureus, and Propionibacterium acnes, and more Preferably, it may be induced by Propionibacterium acnes.

The pharmaceutical composition may be an external preparation for skin.

In more detail, the external preparation for skin may be selected from the group consisting of ointment, cream, lotion, liquid, dressing, patch, blister, tape, aerosol, external acid and spray.

In another embodiment of the present invention, the pharmaceutical composition containing the miabemozaban ethyl acetate fraction as an active ingredient is a suitable carrier, excipient, disintegrant, sweetener, coating agent, swelling agent, lubricant, lubricant commonly used in the manufacture of pharmaceutical compositions. Agents, flavoring agents, antioxidants, buffers, bacteriostatic agents, diluents, dispersants, surfactants, binders and lubricants may further include one or more additives selected from the group consisting of.

Specifically, carriers, excipients and diluents are 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 can be used, and solid dosage forms for oral administration include tablets, pills, powders, granules and capsules. Agents and the like, and such solid preparations may be prepared by mixing at least one excipient in the composition, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

According to an embodiment of the present invention, the pharmaceutical composition is intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalation, topical, rectal, oral, intraocular or intradermal. Routes can be administered to a subject in a conventional manner.

The preferred dosage of the miabemozaban ethyl acetate fraction may vary depending on the condition and weight of the subject, the type and extent of the disease, the drug form, the route and duration of administration, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention is not limited thereto, the daily dosage may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, and more specifically 0.1 to 100 mg / kg. The administration may be administered once a day or divided into several administrations, and the scope of the present invention is not limited thereby.

In the present invention, the 'subject' may be a mammal including a human, but is not limited to these examples.

In addition, the present invention can provide a cosmetic composition for preventing or improving acne containing miabemozaban ethyl acetate fraction as an active ingredient.

The cosmetic composition may include a stabilizer, a solubilizing agent, a common auxiliary agent such as vitamins, pigments, and fragrances, and a carrier, in addition to the ethyl acetate fraction of miabemozaban as an active ingredient.

The cosmetic composition may be prepared in any formulation conventionally prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, oils, powder foundations, emulsion foundations, It may be formulated as a wax foundation and spray, but is not limited thereto. More specifically, it can be prepared in the form of a sun cream, soft lotion, converging lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, pack, spray or powder.

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

When the formulation 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 the case of a spray, additionally chlorofluorohydrocarbon, propane / butane Or propellants such as dimethyl ether.

When the formulation is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 -Fatty acid esters of butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.

When the formulation is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline cellulose , Aluminum metahydroxide, bentonite, agar or trakant, and the like.

Hereinafter, examples will be described in detail to help understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Miabemojaban  Ointment manufacturer

One. Miabemojaban  Ethyl acetate ( Sargassum miyabei Yendo  ethyl acetate; SMYEE) Fraction  Preparations

70% ethanol extract of miabemosaban was concentrated in a vacuum concentrator, and distilled water was added to the concentrated concentrate and suspended at 60 ° C for 1 hour. Thereafter, a 70% ethanol suspension of miabemozaban was added to a separatory funnel for fractionation by solvent polarity, and then hexane in a 1: 1 ratio was sufficiently shaken, and then allowed to stand at room temperature for 3 hours. After 3 hours, the layer was separated, the hexane layer on the upper layer was decanted, and concentrated under reduced pressure to complete the hexane fraction, and the process was repeated 3 times.

The hexane layer was decanted and the remaining distilled water layer was added with the next polar solvent, chloroform, shaken sufficiently, and then left at room temperature for 3 hours. After 3 hours, the layer separation was performed, the chloroform layer at the lower layer was decanted, and concentrated by a vacuum concentrator to complete the chloroform fraction, and the above process was repeated 3 times.

After decanting the chloroform layer, add the next polar solvent ethyl acetate to the remaining distilled water layer, shake it sufficiently, and then leave it at room temperature for 3 hours. After 3 hours, the layer is separated and the ethyl acetate layer on the upper layer is decanted and concentrated in a vacuum concentrator. To obtain an ethyl acetate fraction, the process was repeated 3 times.

2. Miabemojaban  Ointment manufacturer

The ointment containing the miabemozaban ethyl acetate fraction (SMYEE) fractionated by the above process was prepared.

0.4 mL of SMYEE stock (25 mg / mL in 70% ethanol) and 1.3 mL of distilled water were added to Beaker A and heated in a beaker with water maintained at a temperature of 60 to 70 ° C by a hot water method.

In addition, an emulsifier mixed with 0.22 g of oil-based jojoba oil, 0.04 g of emulsion, and 0.04 g of olive oil was placed in a beaker B and stirred using a magnetic bar, and heated to a temperature of 60 to 70 ° C by a hot water method.

When jojoba oil and emulsifier were dissolved and mixed, a solution of beaker A containing SMYEE was added to beaker B, and the stirring speed was increased to maximum and mixed for 5 seconds.

Thereafter, the mixture was cooled while stirring slowly, and the concentration was confirmed. After adding 20 μl of Napre, a natural preservative, the mixture was well mixed and stored in a suitable container for refrigeration.

Also, as a control, an ointment not containing SMYEE was prepared in the same manner as above by adding 0.4 mL of 70% ethanol to the miabemozaban ethyl acetate fraction (SMYEE) instead.

Example (5mg / mL) Comparative example Composition SMYEE stock 0.4 mL 0.4 mL of 70% ethanol Distilled water 1.3 mL Oiled Jojoba Oil 0.22g Emulsifying wax 0.04g Olive oil painting 0.04g Final volume 2 mL 2 mL

< Example  2> Confirm anti-acne effect

1. Acne animal model production

1 × 10 ⁶ by using the sterilization in each of the two parts (2 and 3), the syringe as shown in Figure 1 by three equal parts after epilation the whole is allowed to stand for 24 hours after a stabilization period has passed from the lower end of the ear such as a mouse to tail upper end Acne was induced by injecting CFUs / 10 μL of Propionibacterium acnes (P.acnes) acne, and after 4 days, the inflammation of the mouse skin was confirmed.

2. Miabemojaban  Confirmation of changes in the size of inflammation due to application of ointment

4 days after injection of acne bacteria, ointment not containing the control group miabemozaban ethyl acetate fraction (SMYEE) was applied twice daily for 2 days to the skin and other areas of the skin, such as mice, and SMYEE 50 μg ointment was added to the control group. The same method was applied to areas of the skin, such as the mouse, and the size change of the abscess induced by P. acnes was visually observed.

As a result, it was confirmed that the reduction of the inflammatory response was not observed in the control of the site 2 where the SMYEE-free ointment was applied, whereas the size of the abscess was significantly reduced in the site 3 of the SMYEE-containing ointment. .

3. Histopathology ( Histopathological ) analysis

Four days after application of the ointment, skin tissue from three parts was removed from an animal model such as a mouse acne. Each extracted skin tissue was fixed in 10% formalin and then embedded in a paraffin embedder through a series of tissue treatment processes.

Paraffin-embedded tissue was cut to a thickness of 4 μm, tissue sections prepared were stained with hematoxylin & eosin, and then observed with an optical microscope.

As a result, as shown in FIG. 2, it was confirmed that the abscess formation of the skin tissue 3 with SMYEE-containing ointment was significantly suppressed than that of skin No. 2 with SMYEE-containing ointment.

4. Immunohistochemical staining analysis

Four days after application of the ointment, skin tissue from three parts was removed from an animal model such as a mouse acne. Each extracted skin tissue was fixed in 10% formalin and then embedded in a paraffin embedder through a series of tissue treatment processes.

Paraffin-embedded tissue was cut to a thickness of 4 μm and immunochemical staining was performed with a neutrophil-specific antibody (NIMP-R14, neutrophil marker) on a tissue section slide prepared to confirm the distribution of neutrophils.

As a result, as shown in FIG. 3, it was confirmed that the number of neutrophils was increased in the tissue section of the site 2 injected with P. acnes. In the site, it was confirmed that the number of neutrophils was significantly reduced than the tissue of site 2, which was applied with ointment without SMYEE.

Since the specific parts of the present invention have been described in detail above, for those skilled in the art, it is clear that such specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

A pharmaceutical composition for the prevention or treatment of acne containing the miabemozaban ethyl acetate fraction obtained by fractionating an aqueous solution of miabemozaban ethanol with ethyl acetate. delete The pharmaceutical composition for preventing or treating acne according to claim 1, wherein the ethyl acetate fraction of miabemozaban inhibits the formation of abscess or neutrophil penetration induced by acne bacteria. The method according to claim 1, wherein the acne is induced by acne-causing dermatophytes consisting of Staphylococcus epidermis, Staphylococcus aureus, and Propionibacterium acnes. A pharmaceutical composition for preventing or treating acne, which is characterized by being. The method according to claim 1, wherein the pharmaceutical composition is a pharmaceutical composition for preventing or treating acne, characterized in that the external preparation for skin. The method according to claim 5, wherein the external preparation for skin is selected from the group consisting of ointments, creams, lotions, liquids, dressings, patches, blisters, tapes, mists, external acids and sprays. Therapeutic pharmaceutical composition. A cosmetic composition for preventing or improving acne containing the miabemozaban ethyl acetate fraction obtained by fractionating an aqueous solution of miabemozaban ethanol with ethyl acetate.

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