WO2011149259A2 - Composition including compound derived from peach kernel for treating hyperpigmentation skin disorder and whitening skin - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a functional health food composition including(Z)-12,13-dihydroxyoctadec-9-enoic acid, a compound derived from a peach kernel, for preventing or treating hyperpigmentation skin disorders caused by excess melanin production, and to a composition for whitening the skin. The compound derived from a peach kernel of the present invention,(Z)-12,13-dihydroxyoctadec-9-enoic acid, is a very useful invention for the pharmaceutical and cosmetic industries, since the compound is very effective for skin-whitening activity and effectively inhibits alpha glucosidase and melanin production, thereby preventing, treating, or improving hyperpigmentation skin disorders caused by excess melanin production.

Description

Composition for the treatment of skin hyperpigmented disease and skin whitening containing compound derived from doin

The present invention relates to a composition for the treatment of skin hyperpigmentation disease and skin whitening containing a phosphorus-derived compound, and more specifically, (Z) -12,13-dihydroxyoctadec-9-enoic acid [ (Z) -12,13-dihydroxyoctadec-9-enoic acid] relates to a pharmaceutical composition and a nutraceutical composition for preventing or treating skin hyperpigmentation disease caused by melanin excess formation, and a composition for skin whitening.

Melanin, which determines human skin color, is made from skin cells called melanocytes and migrates to epidermal cells called keratinocytes. Here, melanin plays an important role in forming a cap-like structure around the nucleus to protect genes from ultraviolet rays and to remove free radicals to protect intracellular proteins.

Melanogenesis refers to the biosynthesis of melanin pigments in melanocytes, which are regulated by intracellular and extracellular environments. This melanogenesis is triggered by tyrosinase, which oxidizes tyrosine to dopaquinone, which continues to convert to a pale yellow / red compound, pheomelanin, and light brown to black eumelanin. do. In the skin, melanin affects pigmentation and protects the skin from ultraviolet and other causes of skin disease, while abnormal accumulation of melanin may also cause spots and freckles.

Skin hyperpigmentation diseases caused by melanin hyperplasia include genetic skin disorders such as alum, Badenburg syndrome, acquired skin disorders such as post-inflammatory white nasal mucus, unexplained droplet melaninosis, blemishes, medication Skin diseases caused by, for example, minocycline, bleomycin, busulfan, zidovudine, and skin diseases transmitted through infection, such as stumps.

There are no enzymes that break down melanin in the body, but when keratinocytes break off the epidermis, they are removed by falling off the skin with melanin. However, if more melanin is produced than necessary, it causes hyperpigmentation such as blemishes, freckles, and spots, resulting in cosmetically bad results. In addition, as the leisure population increases, more people enjoy working outside, and the demand to prevent melanin pigmentation due to ultraviolet rays is increasing. Therefore, it was necessary to develop a whitening agent that prevents excessive melanin production and many efforts have been made.

Until now, the development of whitening agents has been mainly focused on finding substances that reduce the amount of melanin by inhibiting the activity of tyrosinase, an essential and most important enzyme in melanin biosynthesis. Recent studies on synthetic whitening agents have explored the use of alkoxy benzoates and alkoxy cinnamate derivatives, and hydroxamic acid. Other studies have shown derivatives of known tyrosinase inhibitors, for example 5-[(3-aninopropyl) phosphinooxy] -2- (hydroxymethyl) -4 H- pyran-4-one {5- [ (3-aminopropyl) phosphinooxy] -2- (hydroxymethyl) -4 H- pyran-4-one}, and N-hydroxy-cinnamoyl pen alkyl- the focal point in the synthesis of amides (N -hydroxycinnamoylphenalkyl-amide) derivative tailored there was.

The whitening agents developed in this way include kojic acid, arbutin, glutathione, vitamin A, and vitamin C. However, the whitening agent does not have a satisfactory whitening effect. Therefore, it is an urgent time to develop a whitening agent that is more effective and safer than ever.

Tyrosinase, an enzyme that synthesizes melanin, is a glycoprotein and is produced through glycosylation, a glycoprotein synthesis in vivo. If there is a problem in the glycosylation process, and the abnormal part of the tyrosinase sugars, the tyrosinase cannot move to the melanosome, a place of intracellular melanin biosynthesis, and thus the melanin production cannot be achieved.

Many enzymes are involved in the glycosylation process, an important enzyme of which is glucosidase. If the enzyme can inhibit the enzymatic activity, it can inhibit the melanin biosynthesis and thus bring about a whitening effect.

 In this regard, Korean Patent Application No. 10-2004-0025053 'Skin Whitening Composition Containing Actinomycetes Culture Extracts' includes actinomycetes strains or Actinomycetes culture extracts, which are enzymes involved in melanin biosynthesis. Glycosylation of tyrosinase by inhibiting the enzymatic activity of beta-glucosidase, an important enzyme involved in the process of glycosylation to synthesize tyrosinase It is disclosed that the whitening effect is exhibited by inhibiting melanin production by inhibiting.

Peach Kernel (Persicae Semen) is a seed of Prunus persica Batsch or Prunus davidiana Maximowicz (Rosaceae). Seeds are used as herbal medicine and are called Persicae Semen.

Doin is a blood clot (血 結), blood, anti-inflammatory, analgesic, menstrual irregularity, constipation is effective in improving blood circulation. Generally, the fruit of the radish tree belonging to the rosaceae is collected after maturation to remove the pulp and core grains, and only the seedlings are dried and dried to make powder.The ingredients are 40 ~ 50% fatty oil, 3.49% cyanide glycoside, 3.8% prunasin, 0.38% emusin , And anticoagulant active substance, and has anticoagulant action and weak hemolysis action to release blood stiffness.瘀血 腫痛) to loosen (Chinese medicine dictionary, p1788) has the effect of turning the blood well. In addition, bleeding, expectoration and lubrication, such as pharmacological action.

The effects of doin from recent scientific research have shown that it protects against cytotoxicity of normal human keratinocytes and fibroblasts due to UV (Archives of Phamacial Research, 2000, 23 (4), It has been reported to contain inhibitors against mutagens (Yakugaku Zasshi-J, Pharmaceutical Society of Japan, 1992, 112 (12), 934-9), hypocholesterolemic mice Has been reported to lower cholesterol levels (Kor. J. Pharmacogn., 1990, 21 (2), 153-157) and scavenging oxygen radicals (Yakugaku Zasshi-J. Pharmaceutical Society of Japan, 1985, 105 (9), 895-901).

Doin extract is a whitening agent in a mixed form of various herbal medicines, for example, Korean Patent Application No. 10-2007-0082003 'Cosmetic Composition Containing Extracts and Fractions of Stein' and Korean Patent Application No. 10-2004-0103894 Preparation of Doin Extract Containing Camperol-3-Galactopyranoside, Camperol-3-0-Alpha-L-Rhamnopyranoside, Camperol-3-0-Beta-L-Glucopyranoside and Its There is an example developed as a 'whitening cosmetic using', but there is no example of developing a whitening substance having a glucosidase inhibitory function or verifying the whitening efficacy using a doin, and no patent is known.

As such, the present inventors have searched for the ability to inhibit the activity of glycosidase enzymes in various plant extracts in order to develop a new whitening substance that is more effective and safer than the whitening agent so far, and the phosphorus extract has a very excellent inhibitory activity of glucosidase enzyme. This was found to be due to (Z) -12,13-dihydroxyoctadec-9-enoic acid and completed the present invention.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating skin hyperpigmented disease caused by melanin overproduction, which contains (Z) -12,13-dihydroxyoctadec-9-enoic acid derived from doin. It is.

Another object of the present invention is to provide a health functional food composition for preventing or improving skin hyperpigmentation disease caused by melanin excess production containing (Z) -12,13-dihydroxyoctadec-9-enoic acid derived from doin. To provide.

Another object of the present invention is to provide a cosmetic composition for skin whitening containing (Z) -12,13-dihydroxyoctadec-9-enoic acid derived from phosphorus as an active ingredient.

An object of the present invention as described above is to prepare an extract from the indoin, and to prepare a fraction from the extract again, after deriving a fraction excellent in the alpha glucosidase inhibitory effect and melanin production inhibitory effect of these fractions, Achieved by identifying chemical structures.

The present invention relates to skin hyperpigmentation due to excessive production of melanin containing (Z) -12,13-dihydroxyoctadec-9-enoic acid of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for preventing or treating a disease is provided.

Formula 1

The present invention also relates to skin overgrowth due to excessive production of melanin containing (Z) -12,13-dihydroxyoctadec-9-enoic acid of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. It provides a health functional food composition for preventing or improving plastic diseases.

The present invention also provides a cosmetic composition for skin whitening containing (Z) -12,13-dihydroxyoctadec-9-enoic acid of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. .

In the present invention, the term "active ingredient" refers to a substance or a group of substances (a medicinal agent whose pharmacologically active ingredient is not known, etc.) which is expected to express the efficacy or effect of the drug directly or indirectly by inherent pharmacological action. It means containing a main component).

In the present invention, "health functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and "functional" means a human body It means the ingestion for the purpose of obtaining a useful effect in health use such as nutrient control or physiological action on the structure and function of.

The compound of formula 1 according to the present invention, melanin by effectively inhibiting the enzymatic activity of glucosidase involved in the glycosylation process of the synthesis process tyrosinase, which is an enzyme that synthesizes melanin in skin cells By inhibiting biosynthesis, the skin whitening activity can be vaporized, and the excessive formation of melamine can be prevented, thereby preventing, ameliorating or treating the skin hyperpigmented disease.

In the present invention, the skin hyperpigmentation disease caused by the melanin hyperplasia is a genetic skin disease, for example, alum, Badenburgh syndrome, acquired skin disease, for example, infectious white nasal mucus, unknown cause water droplet melanin Hypolipidemia, blemishes, skin diseases due to medication, such as minocycline, bleomycin, busulfan, zidovudine, and skin diseases metastasized through infection, such as stumps

Pharmaceutical compositions comprising a compound of formula 1 of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical compositions comprising the compound of formula 1 of the present invention, oral formulations, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods It can be formulated in the form of.

Carriers, excipients and diluents which may be included in pharmaceutical compositions comprising the compound of formula 1 of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate , Gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, 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, sucrose in the extract. Or lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the compound of formula 1 according to the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of formula 1 of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The compound of formula 1 according to the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injections and directly to the skin.

In addition, the compound of formula 1 according to the present invention may be added to food or beverage for the purpose of preventing or improving skin hyperpigmentation disease caused by melanin overproduction. Examples of the food to which the compound of formula 1 may be added include various foods, beverages, gums, teas, vitamin complexes, health functional foods, powders, granules, tablets, capsules, and beverages. At this time, the amount of the compound of formula 1 in the food or beverage may be added to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be added in proportion.

The food composition of the present invention is not particularly limited to other ingredients except for containing the compound of formula 1 as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the compound of formula 1 of the present invention may be used in various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors such as flavoring agents, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like can be used. In addition, the compound of formula 1 of the present invention may contain a flesh for preparing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but it is generally the intent of the present invention to be selected in the range of 0 to about 20 parts by weight per 100 parts by weight.

In the cosmetic composition of the present invention, the composition is characterized in that the formulation of the lotion (skin lotion), lotion, cream, massage cream, essence, pack. The formulations may be prepared using methods known in the art. In addition, the content of the compound of formula 1 in the formulation may also be included within the content range (about 1-10% by weight) conventionally known in the art.

Components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the compound of Formula 1 as an active ingredient, for example, stabilizers, solubilizers, vitamins, pigments and flavorings Conventional adjuvants and carriers are included.

In the present invention, MeOH extract was prepared from Persicae semen , MeOH extract was extracted with EtOAc, and the obtained H ₂ O layer was extracted with n- BuOH / H ₂ O, and each layer obtained therefrom was concentrated under reduced pressure. To give EtOAc, n- BuOH and H ₂ O fractions, respectively.

Subsequently, silica gel column chromatography was performed on the EtOAc fraction to obtain 11 fractions (PPE-1 to PPE-11).

Using the PPE-1 to PPE-11 fractions, alpha-glucosidase recognizes the glucoside moiety of p -nitrophenyl ( p NP) alpha-D-glucoside as a substrate, thereby identifying p NP and alpha-D-glucoside. Cut off by the enzymatic reaction, and the amount of p NP released here was measured by absorbance at 405 nm, thereby indirectly measuring alpha-glucosidase activity. As a result, fractions PPE-1, 2, 4. 10 showed enzymatic activity inhibitory effect (see FIG. 1).

In addition, the melan-a (melan-a) cells, which are immortalized cell lines derived from C57BL / 6 mice, were treated with the fractions, and the PPE-2 fractions showed excellent melanin formation inhibitory activity (see FIG. 2).

Subsequently, ODS c.c. was performed on the PPE-2 fraction which was evaluated to be excellent in inhibiting melanin production, thereby obtaining six fractions (PPE 2-1 to PPE 2-6).

As a result of measuring the effect of inhibiting glucosidase and melanin production for the fraction, it was confirmed that fraction PPE 2-2 had excellent activity (see FIGS. 3 and 4), and the structure thereof was identified (Z) -12, It was confirmed that it is 13-dihydroxyoctadeca-9-enoic acid.

As described above, the phosphorus-derived compound (Z) -12,13-dihydroxyoctadec-9-enoic acid according to the present invention has an excellent effect on inhibiting alpha glucosidase and inhibiting melanin production, resulting in melanin overproduction. The invention can prevent, treat or ameliorate skin hyperpigmentation, and can be very effective for skin whitening activity, which is a very useful invention for the pharmaceutical and cosmetic industries.

1 is a graph showing the results of the evaluation of inhibition of alpha-glucosidase activity of the phosphorus fraction.

Figure 2 is a graph showing the results of evaluation of melanin inhibitory effect in melan-a cells of the indo fraction.

3 is a graph showing the alpha glucosidase inhibitory effect of the PPE 2-2 fraction.

4 is a graph showing the melanin production inhibitory effect of the PPE-2 fraction.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example 1 Preparation of Doin Extract and Separation of Active Substance

5 kg of Persicae semen powder was extracted with MeOH 80% aqueous solution (18 l × 2) twice and then filtered through a filter paper. The obtained filtrate was concentrated under reduced pressure at 45 ° C. to obtain a MeOH extract. The MeOH extract obtained was partitioned and extracted with EtOAc (2 l × 2) / H ₂ O (2 l ), and the obtained H ₂ O layer was again extracted with n- BuOH (2 l × 2) / H ₂ O (3.2 l ). Distributed extraction. Each layer obtained therefrom was concentrated under reduced pressure to give an EtOAc (201 g), n- BuOH (193 g) and H ₂ O fractions, respectively.

Subsequently, silica gel column chromatography (φ 9 × 14 cm, n -hexane-EtOAc = 10: 1 → 7: 1 → 5: 1 → 1: 1 → CHCl ₃ -MeOH = 10: 1 → 7 was used for the EtOAc fraction. : 1 → 5: 1 → 3: 1 → 2: 1 → 1: 1) eleven fractions (PPE-1 to PPE-11) were obtained.

Experimental Example 1 Evaluation of Alpha Glucosidase Inhibition Effect

For the fraction obtained in Example 1, alpha-glucosidase recognizes the glucoside portion of p -nitrophenyl ( p NP) alpha-D-glucoside as a substrate, and p- NP and alpha-D-glucoside are enzymatically reacted. The amount of p NP released here was measured by absorbance at 405 nm, thereby indirectly measuring alpha-glucosidase activity. Sample material and alpha-glucosidase (0.6 unit / ml) were added to 50 mM phosphate buffer (pH 6.8), and 10 mM p NP alpha-D-glycoside solution was added. After the reaction, 1 M Na ₂ CO ₃ was added three times to stop the reaction. The amount of p NP produced in the enzyme reaction was determined by measuring the absorbance at 405 nm.

As a result, as shown in Figure 1, PPE-1, 2, 4. 10 fractions showed the inhibitory effect of enzyme activity.

Experimental Example 2 melanin production inhibitory effect using melan-a (melan-a) cells

Melan-a cells, an immortalized cell line derived from C57BL / 6 mice, were distributed from Dr. Bennett; Cancer Research Center, London, England. Melan-a cells were prepared at 37 ° C., 10 using RPMI 1640 medium containing 10% FBS, 200 nM 12-O-tetradecanoylphorball-13-acetate, 100 mg / L streptomycin, 100 IU / ml penicillin. Incubated under% CO ₂ conditions.

If the melan-a cells grow confluent in the incubator, remove the cells with 0.25% trypsin / EDTA, inject 1x10 ⁵ per well into a 24 well plate, wait 24 hours for the cells to adhere well to the plate, and then with PBS. After washing once, the sample was added to the culture medium containing no FBS and treated with the cells. The sample material was dissolved in DMSO to 1000 times the desired concentration, and then diluted to one thousandth of the amount when used in cells. As a control, a solution obtained by treating DMSO at the same concentration as the sample was used.

After treating the sample material for 3 days as above, the amount of melanin was dissolved in a solution containing 10% DMSO in 1N NaOH and absorbance was measured at 490 nm. The cell lysate was obtained again by the Bradford method and expressed as the amount of melanin per constant protein.

As a result, as shown in Figure 2, PPE-2 fractions showed excellent activity.

Experimental Example 3: Glucosidase Inhibition and Melanin Inhibitory Effects of (Z) -12,13-Dihydroxyoctadeca-9-enoic Acid

ODS cc (φ 3.5 × 11 cm, MeOH-H ₂ O = 7: 1) was performed on the PPE-2 fraction (1.9 g) fraction which was evaluated as having excellent melanin production inhibitory effect in Experimental Example 2, 6 Fractions (PPE 2-1 to PPE 2-6) were obtained.

The fractions were used to measure the effects of glucosidase inhibition and melanin production in the same manner as in Experimental Examples 1 and 2.

As a result, as shown in Figs. 3 and 4, fraction PPE 2-2 [(Compound 1 , 23 mg, TLC (RP-18 F _{254 s} ) R _f = 0.65, MeOH-H ₂ 0 = 5: 1)] It was confirmed that this activity is excellent and its structure was confirmed as follows.

compoundOne[(Z) -12,13-dihydroxyoctadec-9-enoic acid)]: colorless oil, EI / MSm / z: 314 [M]⁺;^OneH-NMR (400 MHz, CDCl₃, δ_H): 5.34 (4H, m.olefinic-H), 2.47-1.42 (22H, CH₂× 11), 0.97 (3H, t,J= 7.4 Hz, terminal methyl);¹³C-NMR (100 MHz, CDCl_3,δ_C): 181.3 (ester), 128.7, 128.5, (olepic-C), 34.9, 31.6, 28.9, 28.7, 28.6, 28.5, 29.3, 29.2, 29.2, 29.1, 29.1, 29.0, 27.3, 27.2, 24.5, 22.3 ( Methylene), 14.3 (terminal methyl).

<Formula 1>

Claims (3)

1. (Z) -12,13-dihydroxyoctadec-9-enoic acid or the pharmaceutically acceptable salt thereof, as an active ingredient, for preventing or treating skin hyperpigmentation disease caused by excessive production of melanin Pharmaceutical compositions.

   <Formula 1>

   
2. Dermal hyperpigmentation disease caused by melanin excess production containing (Z) -12,13-dihydroxyoctadec-9-enoic acid of formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient according to claim 1 Health functional food composition for prevention or improvement.
3. A cosmetic composition for skin whitening comprising (Z) -12,13-dihydroxyoctadec-9-enoic acid or a pharmaceutically acceptable salt thereof according to claim 1 as an active ingredient.

Images