JP2011178754A - SKIN PHOTOAGING PREVENTING OR AMELIORATING COMPOSITION, Ki-67 EXPRESSION INHIBITOR AND ORAL COMPOSITION - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a skin photoaging preventing or ameliorating composition. <P>SOLUTION: A pine bark extract has an effect of preventing or ameliorating photoaging of skin. The pine bark extract also has an effect of suppressing expression of Ki-67 in tumor tissues. The skin photoaging preventing or ameliorating composition containing the pine bark extract as an active ingredient is used with or without addition of various ingredients, as an oral composition such as for foods and drinks or medicines and has an effect of preventing or ameliorating photoaging of skin. The oral composition containing the composition for preventing or ameliorating photoaging of skin can be ingested conveniently and has an effect of preventing or ameliorating photoaging of skin. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a composition for preventing or improving skin photoaging.

In general, skin aging refers to a physiological phenomenon that occurs when physiological aging associated with aging and photoaging due to exposure to ultraviolet rays affect each other. In particular, the relationship between photoaging and skin aging such as rough skin, wrinkles, tarmi, and spots is currently attracting attention. If the skin is exposed to ultraviolet rays for a long period of time, it will not only cause aging of the skin but also cause skin cancer. This is considered to be caused by the fact that DNA damage is caused by causing mutations in the DNA strands of skin cells when the ultraviolet rays of sunlight (particularly UVB) reach the subepidermal layer.

  In order to solve this problem, various compositions for preventing or improving photoaging of the skin have been developed. For example, a skin photoaging preventive agent comprising a phenylethanoid glycoside as an active ingredient (Patent Document 1), a skin photoaging improver comprising a proteoglycan-related substance as a main component (Patent Document 2), kojic acid In addition, a photoaging skin improving cosmetic (Patent Document 3) characterized in that an active ingredient is and / or a derivative thereof is known.

Here, in many tumors such as breast cancer, gastric cancer, colon cancer, uterine cancer, Ki-67, which correlates with tumor malignancy and prognosis such as differentiation degree, vascular invasion and lymph node metastasis, is useful as a cell proliferation marker in tumor tissue. It is known (Non-patent Document 1). Therefore, if the expression of Ki-67 in the tumor tissue can be suppressed, it will greatly contribute to the prevention and improvement of skin photoaging and carcinogenesis due to ultraviolet irradiation.

JP 2008-280271 A JP 2008-195629 A JP 2002-293738 A Arch Dermatol. 2008; 144 (10): 1296-1302

  In recent years, a variety of consumer needs has progressed, and various skin photoaging prevention or improvement compositions have been demanded. However, the prior art alone does not always meet those needs. There has also been a demand for a composition for preventing or improving skin photoaging derived from natural products that can be easily taken.

  As a result of intensive studies, the present inventors have found that a pine bark extract is useful as a material that can be used as a composition for preventing or improving skin photoaging, and have completed the present invention. Furthermore, it discovered that a pine bark extract could suppress the expression of Ki-67 in a tumor tissue, and came to complete this invention.

Specifically, the present invention relates to a composition for preventing or improving skin photoaging using a pine bark extract as an active ingredient and a Ki-67 expression inhibitor containing a pine bark extract as an active ingredient. Preferably, it is an oral composition containing the composition for preventing or improving skin photoaging and a Ki-67 expression inhibitor.

  According to the present invention, the pine bark extract has the effect of preventing or improving skin photoaging. The pine bark extract has the effect of suppressing the expression of Ki-67 in tumor tissue. The composition for preventing or improving skin photoaging comprising pine bark extract as an active ingredient can be used as an oral composition such as foods and drinks and pharmaceuticals as it is or by adding various ingredients. There is an effect of preventing or improving. Furthermore, an oral composition containing a composition for preventing or improving skin photoaging can be easily taken and has the effect of preventing or improving skin photoaging.

It is the figure which showed the period from the ultraviolet irradiation start, and the change of thickening of a mouse skin. It is the figure which showed the period from the ultraviolet irradiation start, and the change of the elasticity of mouse skin. It is the figure which showed the period from the ultraviolet irradiation start, and the change of the tumor number of a mouse | mouth skin. It is the figure which showed the wrinkle formation score of the mouse | mouth of each group after progress for 20 weeks.

  Hereinafter, embodiments of the present invention will be described. In addition, this invention is not limited by description of the below-mentioned embodiment, A various change is possible within the range of description in a claim.

  The pine bark extract used in the present invention contains proanthocyanidins as one of the main active ingredients. Proanthocyanidins refer to a group of compounds composed of a condensation polymer having a degree of polymerization of 2 or more having flavan-3-ol and / or flavan-3,4-diol as a constituent unit. Proanthocyanidins are powerful antioxidants produced by plants and are intensively contained in plant leaves, bark, fruit peels and seeds. This proanthocyanidin is a substance that cannot be produced in the human body.

  The pine bark extract contains a condensation polymer having a degree of polymerization of 2 or more as proanthocyanidins. In particular, proanthocyanidins containing a large amount of a condensation polymer having a low degree of polymerization are desirable. The condensation polymer having a low polymerization degree is preferably a condensation polymer having a polymerization degree of 2 to 30 (2 to 30 mer), more preferably a condensation polymer having a polymerization degree of 2 to 10 (2 to 10 mer). Further, a condensation polymer having a degree of polymerization of 2 to 4 (2 to 4 mer) is more desirable. Proanthocyanidins that are condensation polymers having a degree of polymerization of 2 to 4 (2 to 4 mer) are particularly easily absorbed into the body. In the present specification, a polymer having a degree of polymerization of 2 to 4 is referred to as oligomeric proanthocyanidin (hereinafter referred to as “OPC”).

  As the pine bark extract, a ratio of 15% by mass, preferably 20% by mass or more, more preferably 30% by mass of a condensation polymer having a degree of polymerization of 2 to 4 (2 to 4 mer; that is, OPC). An extract containing a pentamer or more proanthocyanidins in a proportion of 10% by mass or more, desirably 15% by mass or more is desirable.

  The pine bark extract may further contain catechins. The catechins are desirably contained in a proportion of 3% by mass or more, more desirably 5% by mass or more. Catechin is a general term for polyhydroxyflavan-3-ol. As catechins, (+)-catechin (referred to as catechin in a narrow sense), (−)-epicatechin, (+)-gallocatechin, (−)-epigallocatechin, epigallocatechin gallate, epicatechin gallate, and afzelechin Etc. are known. In addition to (+)-catechin, gallocatechin, afzelekin, and (+)-catechin or 3-galloyl derivatives of gallocatechin have been isolated from pine bark extract.

  The catechins are preferably contained in the pine bark extract at 3% by mass or more. More desirably, the pine bark extract containing 20% by mass or more of OPC and 10% by mass or more of pentamer or more proanthocyanidins preferably contains 3% by mass or more of catechins. For example, when the catechin content of the pine bark extract is less than 3% by mass, the catechins may be added so that the content is 3% by mass or more. A pine bark extract containing 3% by mass or more of catechins, 20% by mass or more of OPC, and 10% by mass or more of proanthocyanidins of pentamer or more is desirable.

Pine bark extracts used in the present invention include Pinus Martina, larch, Japanese black pine, Japanese red pine, Japanese pine, Japanese pine, Korean pine, Japanese pine, Ryukyu pine, Utsukushima, Japanese pine, white pine, Canadian Quebec region Bark extracts such as Aneda are desirable. Among them, a bark extract of French coastal pine is desirable.

  French coastal pine is a marine pine that grows on the Atlantic coast of southern France. The bark of this French coastal pine contains organic acids and other physiologically active components in addition to containing proanthocyanidins, which are flavonoids, as main components. This main component, proanthocyanidins, is known to have a strong antioxidant action to remove active oxygen.

  The pine bark extract is obtained by extracting pine bark with water or an organic solvent. When water is used, warm water or hot water is used. As an organic solvent used for extraction, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, butane, acetone, hexane, cyclohexane, propylene glycol, hydrous ethanol, hydrous propylene glycol, ethyl methyl ketone, Organic solvents that are acceptable for the production of food or drugs such as glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible oils and fats, 1,1,1,2-tetrafluoroethane, 1,1,2-trichloroethene Preferably used. These water and organic solvents may be used alone or in combination. In particular, hot water, hydrous ethanol, hydrous propylene glycol and the like are preferably used.

  The extraction method from pine bark for obtaining the pine bark extract is not particularly limited, and for example, a warm extraction method, a supercritical fluid extraction method, or the like is used.

  The supercritical fluid extraction method is a method of performing extraction using a supercritical fluid that is a fluid that exceeds the critical point (critical temperature, critical pressure) of a gas-liquid substance. As the supercritical fluid, carbon dioxide, ethylene, propane, nitrous oxide (laughing gas) or the like is used, and carbon dioxide is preferably used.

  In the supercritical fluid extraction method, an extraction process for extracting a target component with a supercritical fluid and a separation process for separating the target component and the supercritical fluid are performed. In the separation step, any one of extraction separation by pressure change, extraction separation by temperature change, and extraction separation using an adsorbent / absorbent may be performed.

  Moreover, you may perform supercritical fluid extraction by the entrainer addition method. In this method, for example, ethanol, propanol, n-hexane, acetone, toluene, other aliphatic lower alcohols, aliphatic hydrocarbons, aromatic hydrocarbons, and ketones are added to the extraction fluid in an amount of 2 to 20 W / V. %, And by performing supercritical fluid extraction using this fluid, the solubility of the target extract such as OPC and catechins in the extraction solvent is drastically increased, or the separation selectivity is enhanced. This is a method for obtaining an efficient pine bark extract.

  Since the supercritical fluid extraction method can be operated at a relatively low temperature, it can be applied to substances that are altered or decomposed at high temperatures, the advantage that the extraction fluid does not remain, and the recycling of the solvent is possible. There is an advantage that the process and the like can be omitted, and the process becomes simple.

  Further, the extraction method from pine bark for obtaining a pine bark extract may be performed by a method such as a liquid carbon dioxide batch method, a liquid carbon dioxide reflux method, a supercritical carbon dioxide reflux method, etc., in addition to the above extraction method. Good.

  The extraction method from the pine bark for obtaining the pine bark extract may be a combination of a plurality of extraction methods. By combining a plurality of extraction methods, it becomes possible to obtain pine bark extracts having various compositions.

  It is safe to purify the pine bark extract obtained by the above-described extraction by ultrafiltration or a column method or a batch method using an adsorptive carrier (Diaion HP-20, Sephadex-LH20, chitin, etc.). From the viewpoint of sex.

  The amount of the composition for preventing or improving skin photoaging used in the present invention is not particularly limited, but 2 mg or more of pine bark extract per day is preferable, and if it is 2 mg or less, a sufficient effect cannot be expected. The administration may be divided into several times a day. Further, a pine bark extract manufactured by Toyo Shinyaku Co., Ltd. can be preferably used.

  The amount of Ki-67 expression inhibitor used in the present invention is not particularly limited, but 2 mg or more of pine bark extract per day is preferable, and if it is 2 mg or less, a sufficient effect cannot be expected. The administration may be divided into several times a day. Further, a pine bark extract manufactured by Toyo Shinyaku Co., Ltd. can be preferably used.

  The composition for preventing or improving skin photoaging and Ki-67 expression inhibitor used in the present invention should be used as oral compositions such as foods and beverages and pharmaceuticals and cosmetics as they are or with various components added. Can do. The amount of pine bark extract added to oral compositions and cosmetics is not particularly limited, but 2 mg or more is preferable per day per individual, and if it is 2 mg or less, a sufficient effect cannot be expected. The administration may be divided into several times a day.

  As foods and drinks, the composition for skin photoaging prevention or improvement described in the present invention and the Ki-67 expression inhibitor are used as they are, or various nutritional components are added to the foods, for example, powdered / granular forms.・ Granular, Liquid, Paste, Cream, Tablet, Capsule, Caplet, Soft Capsule, Tablet, Bar, Plate, Block, Pill, Solid, Gel, Jelly, Gummy, It can be molded into a wafer, biscuit, bowl, chewable, syrup, stick or the like and provided as a food material. Further, it may be used by adding to water, milk, soy milk, fruit juice drink, whey drink, soft drink, green juice, yogurt or the like. Further, it may be added to health food or food for specified health use.

  As pharmaceuticals, for example, powders, granules, granules, liquids, tablets, capsules, caplets, soft capsules, tablets, rods, plates, blocks, pills, etc. are provided as pharmaceuticals can do.

The oral composition of the present invention, when taken orally, suppresses the expression of Ki-67 in the tumor tissue and improves skin symptoms (for example, rough skin, wrinkles, tarmi, stains, tumors) due to photoaging. There is an effect.

  As cosmetics, for example, lotion, cosmetic cream, milky lotion, pack, hair tonic, hair cream, shampoo, hair rinse, treatment, body shampoo, facial cleanser, soap, foundation, white powder, lipstick, lip gloss, blusher, eye shadow It is used as hair conditioner, hair conditioner, aqueous ointment, oily ointment, eye drops, eye wash, ship, gel, etc. The cosmetics of the present invention suppresses the expression of Ki-67 in tumor tissue by applying or spraying on the skin or the like, thereby reducing skin symptoms due to photoaging (for example, rough skin, wrinkles, tarmi, stains, tumors) and the like. There is an action to improve.

  Examples of the present invention will be described below. It should be noted that the present invention is not construed as being limited to the examples described below, and various modifications can be made within the scope of the claims. The pine bark extract made by Toyo Shinyaku Co., Ltd. was used as the pine bark extract.

A 4-week-old HRM melanin-containing hairless mouse (weight: about 15 to 20 g, manufactured by Japan SLC Co., Ltd.), a normal group that does not irradiate UVB, a control group that only irradiates UVB, and a pine bark extract that is irradiated with UVB The test food low dose group administered orally at a low dose and the test meal high dose group administered orally with a high dose of pine bark extract by UVB irradiation were divided into 4 groups of 8 (Table 1).
(Table 1)
The mice in each group except the normal group were irradiated with a UVB lamp (T-15M, 15W, 312 nm, Ieda Boseki Co., Ltd.) every other day for 20 weeks. UVB irradiation started from a low volume and gradually increased the dose. Specifically, 1 week 36 mJ / ^cm 2, 2 to 4 week 54 mJ / ^cm 2, 4 to 7 week 72 mJ / ^cm 2, 7 to 10 week 108mJ / ^cm 2, 10~13 th week is 144mJ / ^cm 2, 13~20 week was irradiated with 168mJ / cm ^2. In parallel, an aqueous solution containing a pine bark extract was orally administered twice daily in the morning and evening to the group taking the test meal. The amount of the pine bark extract administered at one time is as shown in Table 1, and is 60 mg / kg in the test meal low dose group and 200 mg / kg in the test meal high dose group. That is, for example, in the case of a mouse having a body weight of 15 g, the amount of pine bark extract administered at a time is 0.9 mg in the test meal low-dose group and 3.0 mg in the test meal high-dose group. In the normal group and the control group, only water was administered.

(Example 1: Evaluation of skin photoaging)
Skin photoaging was evaluated for changes in skin thickening and elasticity, skin tumor number, and wrinkle formation. The result was tested by the Turkey-Kramer method using statistical processing software, and p <0.05 was judged to be significant.

Skin thickening and elasticity (skin elongation) and skin tumor number were measured every week. FIG. 1 shows the results of thickening, FIG. 2 shows the results of elasticity, and FIG. 3 shows the results of the number of tumors.

For skin wrinkles, wrinkle formation was scored in 6 stages using photographs taken at 8th, 12th, 16th and 20th weeks. That is, normal skin with no wrinkles was grade 0, those with shallow wrinkles were grade 2, those with rough wrinkles were grade 4, and those with deep wrinkles were grade 6. The results are shown in FIG.

As shown in FIGS. 1 to 4, by oral administration of pine bark extract (60 mg / kg, 200 mg / kg), the skin thickening is from the 8th week onward (FIG. 1), and the skin elasticity is from the 17th week onward. (FIG. 2) In skin tumors, a significantly low value was observed after 18 weeks of administration (FIG. 3). Regarding wrinkle formation, a significant low value was observed at 20 weeks after oral administration of pine bark extract (200 mg / kg) (FIG. 4). Further, in the pathological evaluation by HE staining, Azan staining, and Fontana-Matson staining, a significant suppression of the thickening of the epidermis layer by UVB irradiation and a significant suppression of an increase in melanin granule formation were confirmed. From the above results, it was shown that skin thickening, reduced elasticity, wrinkle formation, and carcinogenesis (skin tumors), which are various symptoms of skin photoaging, can be prevented or improved by pine bark extract.

(Example 2: Examination of mechanism)
The skin tissue pieces of each group of mice in Table 1 were fixed with 10% formalin buffer, and pathological sections were prepared according to a conventional method. Immunostaining of skin pathology is performed according to a conventional method using an anti-mouse Ki-67 rat monoclonal antibody (DakoCytomation), an anti-8-OHdG antibody (Japan Aging Control Laboratories), and an apoptosis detection kit (Wako Pure Chemical Industries, Ltd.). The number of Ki-67 positive cells, the number of 8-OHdG positive cells and the number of apoptotic cells were measured.

The results are shown in Table 2. It was revealed that Ki-67 positive cells increased in the basal layer by UVB irradiation, and the cells were proliferating. Oral administration of pine bark extract (60 mg / kg, 200 mg / kg) significantly suppressed the increase in Ki-67-expressing cells. In addition, the number of 8-OHdG positive cells was increased by UVB irradiation, and it was found that the cells were proliferating. A tendency to suppress the increase in 8-OHdG-expressing cells was observed by oral administration of pine bark extract (60 mg / kg, 200 mg / kg). It was found that UVB irradiation caused skin cells to undergo apoptosis. Apoptosis was significantly suppressed by oral administration of pine bark extract (60 mg / kg, 200 mg / kg). From the above results, it was shown that the pine bark extract has an effect of suppressing the expression of Ki-67 and 8-OHdG in tumor cells.
(Table 2)

In addition, the expression levels of matrix metalloproteinase (MMP) -2 and MMP-9 were measured using the mice of each of the above groups. Although MMP-2 and MMP-9 increased by UVB irradiation, suppression of the expression level by oral administration of a pine bark extract could not be confirmed. Therefore, it is presumed that the mechanism of the improvement effect of skin photoaging by the pine bark extract is not the suppression of the expression of MMP-2 and MMP-9.

  In summary, the pine bark extract has the effect of preventing or improving skin photoaging. In addition, the pine bark extract has an effect of suppressing the expression of Ki-67 and 8-OHdG in the tumor tissue. Therefore, the pine bark extract has the effect of preventing or improving skin photoaging by suppressing the expression of Ki-67 and 8-OHdG in tumor cells. Moreover, the pine bark extract has the effect of preventing cell damage (carcinogenesis) due to UVB irradiation from the result that the pine bark extract suppressed the increase in apoptosis due to UVB irradiation.

  According to the present invention, the pine bark extract has the effect of preventing or improving skin photoaging. The pine bark extract has the effect of suppressing the expression of Ki-67 in tumor tissue. The composition for preventing or improving skin photoaging comprising pine bark extract as an active ingredient can be used as an oral composition for foods and drinks, pharmaceuticals, etc. as it is or by adding various ingredients. It has the effect of preventing or improving various symptoms. Furthermore, the composition for preventing or improving skin photoaging can be easily taken as an oral composition such as foods and drinks and pharmaceuticals.

Claims (4)

A composition for preventing or improving skin photoaging, comprising a pine bark extract as an active ingredient. A Ki-67 expression inhibitor comprising pine bark extract as an active ingredient. An oral composition containing the composition for preventing or improving skin photoaging according to claim 1. The composition for oral administration containing the Ki-67 expression inhibitor of Claim 2.