TWI636796B - EXTERNAL COMPOSITION FOR SKIN CONTAINING GINSENOSIDE Rg3 - Google Patents

EXTERNAL COMPOSITION FOR SKIN CONTAINING GINSENOSIDE Rg3 Download PDF

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TWI636796B
TWI636796B TW103114697A TW103114697A TWI636796B TW I636796 B TWI636796 B TW I636796B TW 103114697 A TW103114697 A TW 103114697A TW 103114697 A TW103114697 A TW 103114697A TW I636796 B TWI636796 B TW I636796B
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ginsenoside
skin
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effect
active ingredient
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TW201519909A (en
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金東泫
柳權烈
李沃澯
廉明勳
曺濬喆
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愛茉莉太平洋股份有限公司
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    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A—HUMAN NECESSITIES
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    • A—HUMAN NECESSITIES
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/02—Local antiseptics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/006—Antidandruff preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth

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Abstract

本發明係關於一種以含人參皂苷Rg3為有效成分而可提供改善痤瘡及皮膚問題之效果、皮膚收斂及毛孔收縮效果，亦可提供皮膚氣色改善效果、促進頭髮生長、改善白髮、抗頭屑及防腐效果之組合物。 The present invention relates to a ginsenoside-containing Rg3 as an active ingredient, which can provide the effect of improving acne and skin problems, the effect of skin convergence and pore shrinkage, and can also provide the effect of improving skin color, promoting hair growth, improving white hair, and anti-dandruff. And antiseptic effect composition.

Description

含人參皂苷Rg3的皮膚外用劑組合物 Skin external preparation composition containing ginsenoside Rg3

本發明係關於一種藉由含人參皂苷Rg3而可提供改善痤瘡及皮膚問題之效果、皮膚收斂及毛孔收縮效果，亦可提供皮膚氣色改善效果、促進頭髮生長、改善白髮、抗頭屑及防腐效果之組合物。 The present invention relates to a ginsenoside-containing Rg3, which can provide the effect of improving acne and skin problems, the effect of skin convergence and pore shrinkage, and also the effect of improving skin color, promoting hair growth, improving white hair, anti-dandruff and anti-corrosion Effect composition.

人類之皮膚係一種人體之第一層防護膜，其在溫度及濕度之變化、紫外線、公害物質等外部環境之刺激中起到保護體內之器官之功能，隨著年齡之增長，其經歷各種內外要因之變化。即，在內在要因方面，用以調節新陳代謝之各種荷爾蒙之分泌減少，免疫細胞之功能和細胞之活性降低，從而導致人體所需之免疫蛋白質及人體結構蛋白質之生物合成性減少；在外在要因方面，因臭氧層遭到破壞，太陽光線之到達地表之紫外線之含量增加，並且，隨著環境污染之逐漸加劇，導致自由基及活性氧等增加，從而導致皮膚厚度減少、皺紋增加、彈性下降，亦導致皮膚氣色變得暗沉，經常出現皮膚問題，增加黃褐斑和雀斑以及老年斑，氣色變差膚色變暗等，導致出現各種變化。 Human skin is the first protective film of the human body. It plays a role in protecting the organs of the body during the stimulation of external environments such as temperature and humidity changes, ultraviolet rays, and harmful substances. With the increase of age, it undergoes various internal and external Causes change. That is, in terms of internal factors, the secretion of various hormones used to regulate metabolism is reduced, the function of immune cells and cell activity are reduced, resulting in a decrease in the biosynthesis of immune proteins and body structural proteins required by the human body; in terms of external factors Due to the destruction of the ozone layer, the amount of ultraviolet rays reaching the surface of the sun's rays has increased, and as environmental pollution has gradually intensified, free radicals and active oxygen have increased, resulting in reduced skin thickness, increased wrinkles, and reduced elasticity. As a result, skin complexion becomes dull, skin problems often occur, melasma and freckles and age spots are increased, poor complexion becomes darker, skin tone becomes darker, etc., resulting in various changes.

為了防止因此種皮膚內在及外在要因所導致之皮膚狀態之變化和為了保持健康之皮膚狀態，藉由將提取自已知之各種動物、植物、微生物等之生物活性物質添加至化妝品後使用，從而嘗試改善皮膚狀態。 In order to prevent changes in the skin state caused by internal and external causes of this kind of skin and to maintain a healthy skin state, bioactive substances extracted from various known animals, plants, microorganisms, etc. are added to cosmetics and used. Improve skin condition.

人參皂苷(Ginsenoside)是指存在於人參中的皂苷，皂苷為化學名稱為配糖體(glycoside)之化合物的一種。為了與其它植物皂苷區別，人參皂苷以從人參(ginseng)分離之配糖體(glyco-side)之意思命名。如此，一般以人參皂苷-Rx(ginsenoside-Rx)等來命名。在此，“R”之意思為根(Radix或Root)，“x”之意思為依照在TLC(薄層色譜法)上所表示之點(spot)之移動距離(Rf值)，以從下面向上面為o、a、b、c、d、e、f、g及h等之依序命名。人參皂苷對包括中樞神經系統在內、及於內分泌系統、免疫系統、代謝系統等造成影響，從而在身體功能調整上發揮多樣之效果。至今，從高麗人參(含紅參)，30多種以上之人參皂苷被分離鑑定，各個人參皂苷的藥理作用陸續地被發現。上述人參皂苷彼此作相似之作用或者彼此表示出相反之作用。例如，對於中樞神經系，代表PPD系皂苷之人參皂苷G-Rb1表現抑制之效果，而代表PPT系皂苷之G-Rg1則具有促進之效果。 Ginsenoside refers to the saponin existing in ginseng, and saponin is a kind of compound whose chemical name is glycoside. To distinguish it from other plant saponins, ginsenosides are named after glyco-sides isolated from ginseng. As such, it is generally named after ginsenoside-Rx (ginsenoside-Rx) and the like. Here, "R" means the root (Radix or Root), and "x" means the moving distance (Rf value) according to the point (spot) indicated on the TLC (thin layer chromatography), from below Name them in the order of o, a, b, c, d, e, f, g, h, etc. Ginsenosides affect the central nervous system, as well as the endocrine system, immune system, and metabolic system, and exert various effects on body function adjustment. So far, more than 30 kinds of ginsenosides have been isolated and identified from Korean ginseng (including red ginseng), and the pharmacological effects of each ginsenoside have been discovered successively. The above-mentioned ginsenosides perform similar effects to each other or show opposite effects to each other. For example, for the central nervous system, ginsenoside G-Rb1, which represents PPD-based saponins, has an inhibitory effect, and G-Rg1, which represents PPT-based saponins, has a promoting effect.

人參皂苷依照皂苷糖苷配基(Aglycon)的構造可分別三種為人蔘二醇型人參皂苷(Panaxadiol-type Ginsenosides：Ra1、Ra2、Ra3、Rb1、Rb2、Rb3、Rc、Rd、Rg3)，人蔘三醇型人參皂苷(Panaxatriol-type Ginsenosides：Re、Rg3、Rg1、Rg2)、齊墩果酸型人參皂苷(Oleanolic acid-type Ginsenosides)等。在人參根上人蔘二醇型皂苷之含量以48~60%最多，其次，人蔘三醇型 皂苷之含量佔30~35%，齊墩果酸型皂苷含有微量。 According to the structure of saponin aglycon, three types of ginsenosides are Panaxadiol-type Ginsenosides: Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, and Rg3. Panaxatriol-type Ginsenosides (Re, Rg3, Rg1, Rg2), oleanolic acid-type Ginsenosides, and the like. The content of ginsenoside saponin on ginseng root is 48 ~ 60% at most. Secondly, ginsenoside type The saponin content accounts for 30 ~ 35%, and the oleanolic acid type saponin contains a trace amount.

關於這樣的人參在蒸參之過程當中所生成之少量人參皂 苷之Rg3之藥理作用具有癌症細胞轉移抑制作用、血小板凝聚抑制作用及抗血栓作用、血管弛緩作用、試驗的肝損傷抑制作用、抗癌劑之耐藥性抑制作用等口服用藥理作用之專利，但是至今還未公開以人參皂苷Rg3為有效成分之組合物之改善痤瘡及皮膚問題之效果、皮膚收斂及毛孔收縮效果、皮膚氣色改善效果、促進頭髮生長、改善白髮、抗頭屑及防腐效果之整體效果。 About the small amount of ginseng soap produced by such ginseng during the steaming process The pharmacological effects of glycoside Rg3 have patents for oral pharmacological effects such as cancer cell metastasis inhibition, platelet aggregation inhibition and antithrombotic effects, vasorelaxation effects, experimental liver injury inhibition effects, anticancer drug resistance inhibition effects, etc. However, the effects of a composition containing ginsenoside Rg3 as an active ingredient to improve acne and skin problems, skin astringency and pore shrinkage effects, skin complexion improvement effects, hair growth promotion, gray hair improvement, antidandruff and antiseptic effects have not been disclosed so far. The overall effect.

對此，本發明人發現了人參皂苷Rg3可提供改善痤瘡及皮膚問題之效果、皮膚收斂及毛孔收縮效果，亦可提供皮膚氣色改善效果、促進頭髮生長、改善白髮、抗頭屑及防腐效果，從而完成了本發明。 In response, the present inventors discovered that ginsenoside Rg3 can provide the effects of improving acne and skin problems, skin convergence and pore shrinkage effects, and can also provide skin color improvement effects, promote hair growth, improve white hair, anti-dandruff and anti-corrosion Thus, the present invention has been completed.

因而，本發明之目的在於提供一種藉由含人參皂苷Rg3而表現出可提供改善痤瘡及皮膚問題之效果、皮膚收斂及毛孔收縮效果，亦可提供皮膚氣色改善效果、促進頭髮生長、改善白髮、抗頭屑及防腐效果之皮膚外用劑組合物。 Therefore, the object of the present invention is to provide a ginsenoside-containing Rg3 that exhibits effects that can improve acne and skin problems, skin convergence and pore shrinkage effects, and can also provide skin color improvement effects, promote hair growth, and improve white hair. Skin external preparation composition with antidandruff and antiseptic effect.

為了達到上述目的，本發明提供一種將人參皂苷Rg3，作為有效成分包含於其中之痤瘡改善用皮膚外用劑組合物。 In order to achieve the above-mentioned object, the present invention provides a skin external preparation composition for improving acne containing ginsenoside Rg3 as an active ingredient.

又，本發明提供一種將人參皂苷Rg3作為有效成分包含其中之氣色及膚色改善用皮膚外用劑組合物。 In addition, the present invention provides a skin external preparation composition for improving color and skin tone containing ginsenoside Rg3 as an active ingredient.

又，本發明提供一種將人參皂苷Rg3作為有效成分包含其 中之毛孔收縮用皮膚外用劑組合物。 The present invention also provides ginsenoside Rg3 as an active ingredient. Pore shrinkage skin external preparation composition.

又，本發明提供一種含有人參皂苷Rg3之毛髮生長促進用組合物。 The present invention also provides a composition for promoting hair growth containing ginsenoside Rg3.

又，本發明提供一種含有人參皂苷Rg3之白髮預防用組合物。 The present invention also provides a composition for preventing gray hair containing ginsenoside Rg3.

又，本發明提供一種含有人參皂苷Rg3之抗頭屑用組合物。 The present invention also provides an antidandruff composition containing ginsenoside Rg3.

又，本發明提供一種含有人參皂苷Rg3之天然防腐劑組合物。 The present invention also provides a natural preservative composition containing ginsenoside Rg3.

本發明之組合物藉由含人參皂苷Rg3，可提供改善痤瘡及皮膚問題之效果、皮膚收斂及毛孔收縮效果，亦可提供皮膚氣色改善效果、促進頭髮生長、改善白髮、抗頭屑及防腐效果。 The composition of the present invention, by containing ginsenoside Rg3, can provide the effect of improving acne and skin problems, the effect of skin convergence and pore shrinkage, and also the effect of improving skin color, promoting hair growth, improving white hair, anti-dandruff and anti-corrosion effect.

根據本發明之組合物將人參皂苷Rg3(ginsenoside Rg3)作為有效成分包含其中。 The composition according to the present invention contains ginsenoside Rg3 (ginsenoside Rg3) as an active ingredient.

於本發明中所使用之人參皂苷Rg3具有以下化學式1之結構。 The ginsenoside Rg3 used in the present invention has a structure of the following Chemical Formula 1.

[化學式1] [Chemical Formula 1]

本發明之人參皂苷Rg3可提取自植物，亦可依照本領域公開方法合成而使用，亦可使用商業市面有售之人參皂苷Rg3。又，人參皂苷Rg3可自人參提取物中獲取。此時，所使用之人參之種類不受特殊限制，可使用水參、紅參、白參、太極參、尾參等。又，上述人參提取物包括所有藉由自人參浸出、煎出而獲取之浸出液、藉由對浸出液之部分或全部再次進行濃縮而獲取之濃縮物或者藉由將上述之濃縮物再次進行乾燥而製備之沉體、煎劑、酊劑、流體提取物及含於人參中而發揮主要效果之化學物質，還包括其植物本身，提取物可取自莖、根、葉、花、果等人參之所有部分，不限定為某一特定部分之提取物。又，自人參提取物提取人參皂苷Rg3之方法可使用公開之方法。 The ginsenoside Rg3 of the present invention can be extracted from plants, can also be synthesized and used according to the methods disclosed in the art, and ginsenoside Rg3 commercially available can also be used. Ginsenoside Rg3 can be obtained from ginseng extract. At this time, the type of ginseng used is not particularly limited, and water ginseng, red ginseng, white ginseng, Taiji ginseng, and tail ginseng can be used. In addition, the above-mentioned ginseng extract includes all extracts obtained by leaching and decocting from ginseng, concentrates obtained by re-concentrating part or all of the extracts, or preparations by re-drying the above-mentioned concentrates The sinking body, decoction, tincture, fluid extract and chemicals contained in ginseng which exert its main effect, including the plant itself, the extract can be taken from all parts of ginseng such as stems, roots, leaves, flowers, fruits, etc. , Not limited to extracts from a specific part. In addition, as a method for extracting ginsenoside Rg3 from a ginseng extract, a published method can be used.

具體而言，藉由本領域眾所周知之方法以水或有機溶劑於人參中提取人參提取物後，由此可分離出上述人參皂苷Rg3。本發明使用之有機溶劑可選自包括乙醇、甲醇、丁醇、***、乙酸乙酯、氯仿及該些有機溶劑和水之混合溶劑之組之一種，優選地，使用80%乙醇。此時，優選地，提取溫度為10℃~80℃，可提取3小時~24小時。若超出上述提取溫度及提取時間，則將導 致提取效率下降或發生成分之變化。 Specifically, after the ginseng extract is extracted from ginseng with water or an organic solvent by a method well known in the art, the aforementioned ginsenoside Rg3 can be isolated. The organic solvent used in the present invention may be selected from the group consisting of ethanol, methanol, butanol, diethyl ether, ethyl acetate, chloroform, and a mixed solvent of these organic solvents and water. Preferably, 80% ethanol is used. At this time, preferably, the extraction temperature is 10 ° C to 80 ° C, and extraction can be performed for 3 hours to 24 hours. If the above extraction temperature and extraction time are exceeded, As a result, the extraction efficiency decreases or the composition changes.

優選地，於本發明之組合物中，上述人參皂苷Rg3之含量為組合物總重量之0.001重量%~50重量%。若上述有效成分之含量不足0.001重量%，則上述成分之效能、效果微弱；若上述有效成分之含量超過50重量%，則導致皮膚安全性或劑型上發生問題。 Preferably, in the composition of the present invention, the content of the ginsenoside Rg3 is 0.001% to 50% by weight based on the total weight of the composition. If the content of the active ingredient is less than 0.001% by weight, the efficacy and effect of the component are weak; if the content of the active ingredient exceeds 50% by weight, problems in skin safety or dosage form may occur.

本發明之組合物可作為痤瘡改善用皮膚外用劑組合物而使用，其抗菌效果優秀，尤其對痤瘡致病細菌之抗菌效果突出，又，其提供抗炎效果。 The composition of the present invention can be used as a skin external preparation composition for improving acne, and has excellent antibacterial effect, especially has outstanding antibacterial effect on acne-causing bacteria, and also provides anti-inflammatory effect.

本發明之組合物可作為氣色及膚色改善用皮膚外用劑組合物而使用，其適用於皮膚後，藉由擴張毛細血管和促進血液循環來確保皮膚順利吸收營養成分，從而遏制老化，其氣色及膚色改善效果卓越。 The composition of the present invention can be used as a skin external preparation composition for improving color and complexion. After being applied to the skin, it can ensure smooth absorption of nutrients by expanding the capillaries and promoting blood circulation, thereby suppressing aging. Improves skin tone.

本發明之組合物可作為毛孔收縮、皮脂調節及皮膚問題改善用皮膚外用劑組合物而使用，其適用於皮膚後，遏制因過剩而分泌之皮脂，促進活性氧之消除和膠原蛋白之合成，從而收縮毛孔，其以炎症因子之表現減少而遏制皮膚問題之效果突出。 The composition of the present invention can be used as a skin external preparation composition for pore shrinkage, sebum regulation, and improvement of skin problems. After being applied to the skin, it suppresses sebum secreted due to excess, promotes elimination of active oxygen and synthesis of collagen, As a result, the pores are contracted, and the effect of curbing skin problems is reduced by the reduction of the expression of inflammatory factors.

本發明之組合物可作為毛髮生長促進用組合物而使用，其藉由促進休止期毛髮週期進入成長期毛髮週期來促進毛髮之生長和新毛髮之生成，亦可確保已有毛髮健康生長，預防和遏制掉發現象或毛髮變稀變細之狀態。 The composition of the present invention can be used as a composition for promoting hair growth, which promotes the growth of hair and the generation of new hair by promoting the hair cycle from the resting phase to the hair growth phase, and can also ensure the healthy growth of existing hair and prevent And curb the state of finding elephants or thinning hair.

本發明之組合物可作為白髮預防用組合物而使用，其藉由增加黑素細胞(melanocyte)之小眼畸形相關轉錄因子(MITF)表現來活化黑素細胞並促進黑色素合成，從而提供提前 預防白髮之生成和促進黑髮生長之效果。 The composition of the present invention can be used as a composition for preventing gray hair, and it can provide melanin cells by increasing the expression of small eye malformation-related transcription factor (MITF) of melanocytes, and promoting melanin synthesis, thereby providing advancement. Prevent the formation of white hair and promote the growth of black hair.

本發明之組合物可作為抗頭屑用皮膚外用劑組合物而使用，其有效***於毛髮和頭皮中積累之毒素，淨化頭皮，遏制頭屑細菌之增殖和成長，從而可預防頭皮炎症反應，又，其遏制活性氧之生成及作用之抗氧化效能突出，從而可提供舒緩和增強頭皮、強化固有防禦能力之效果。 The composition of the present invention can be used as an anti-dandruff skin external preparation composition, which can effectively excrete toxins accumulated in hair and scalp, purify the scalp, suppress the proliferation and growth of dandruff bacteria, and thereby prevent scalp inflammation reactions. In addition, its outstanding anti-oxidant effect in inhibiting the generation and action of active oxygen can provide the effect of soothing and strengthening the scalp and strengthening the inherent defense ability.

本發明之組合物可作為天然防腐劑組合物而使用，其含有天然成分，其防腐效果出色，提供對人體無害之效果。 The composition of the present invention can be used as a natural preservative composition. It contains natural ingredients, has excellent antiseptic effect, and provides harmless effects to human body.

根據本發明之組合物可藉由含化妝品學或皮膚科學允許之介質或機制而被劑型化。其作為適合於局部適用之所有劑型，例如，能夠以溶液、凝膠、固體、攪拌無水生成物、將乳狀分散於水狀而獲得之乳液、懸浮液、微乳液、微膠囊、微細顆粒球或離子型(脂質體)(Liposomes)及非離子型之水泡(vesicular)分散劑之形態而提供，或者亦能夠以乳霜、化妝水、乳液、粉體、軟膏、噴霧或遮瑕膏之形態而提供。或者亦能夠以泡沫(Foam)形態或更含有被壓縮之推進劑之氣溶膠組合物之形態而使用。該些組合物可按該領域之通常之方法製備。 The composition according to the present invention can be formulated by including a cosmetically or dermatologically acceptable medium or mechanism. It is suitable for all dosage forms suitable for topical application, for example, emulsions, suspensions, microemulsions, microcapsules, and fine particle spheres that can be obtained as solutions, gels, solids, stirred anhydrous products, and dispersion of milky water. Or ionic (liposomes) and non-ionic vesicular dispersants, or they can be in the form of creams, lotions, lotions, powders, ointments, sprays or concealers provide. Alternatively, it can be used in the form of a foam (foam) or an aerosol composition containing a compressed propellant. These compositions can be prepared according to the usual methods in this field.

尤其，本發明之皮膚外用劑組合物，若作為抗頭屑、育髮或白髮預防用途而被使用，則可劑型化為頭皮及毛髮用組合物，劑型不受特殊限定，例如，可劑型化為護髮素、毛髮營養水、柔順劑、護髮膏、洗髮乳、護髮乳、髮乳或頭皮毛髮兩用護理膏等。 In particular, if the skin external preparation composition of the present invention is used as an antidandruff, hair growth or gray hair prevention application, it can be formulated into a composition for scalp and hair, and the dosage form is not particularly limited. For example, it can be a dosage form Turn into conditioner, hair nourishing water, softener, hair cream, shampoo, hair cream, hair cream or scalp hair dual-use care cream.

又，根據本發明之組合物可含有脂肪物質、有機溶劑、溶解劑、濃縮劑、凝膠劑、軟化劑、抗氧化劑、懸浮劑、穩定 劑、發泡劑(foaming agent)、芳香劑、表面活性劑、水、離子型或非離子型乳化劑、填充劑、金屬離子螯合劑(Chelating agents)、螯合劑、防腐劑、維生素、阻斷劑、濕潤劑、必要油、染料、顏料、親水性或親油性活性劑、脂質水泡(vesicular)或如通常用於化妝品之任意其他成分之於化妝品學或皮膚科學領域中通常使用之輔助劑。上述輔助劑之使用量以於化妝品學或皮膚科學領域中通常使用之量為準。 In addition, the composition according to the present invention may contain a fatty substance, an organic solvent, a dissolving agent, a concentrating agent, a gelling agent, a softener, an antioxidant, a suspending agent, and a stabilizer. Agents, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, chelating agents, chelating agents, preservatives, vitamins, blocking agents Agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, vesicular lipids, or any other ingredients commonly used in cosmetics, are commonly used in the field of cosmetics or dermatology. The amount of the above-mentioned adjuvant is based on the amount usually used in the field of cosmetics or dermatology.

又，本發明之組合物為增加皮膚改善效果而可含有皮膚吸收促進物質。 In addition, the composition of the present invention may contain a skin absorption promoting substance in order to increase the skin improving effect.

【實施形態】 [Implementation form]

以下，將例舉試驗例及劑型例，更具體地說明本發明之結構及效果。然而，該些試驗例及劑型例係為了幫助理解本發明而以示例為目的提供，本發明之範疇及範圍並非限定於下述例。 In the following, test examples and formulation examples will be exemplified to more specifically describe the structure and effects of the present invention. However, these test examples and formulation examples are provided for the purpose of understanding the present invention, and the scope and scope of the present invention are not limited to the following examples.

[參考例1]人參皂苷Rg3之準備 [Reference Example 1] Preparation of ginsenoside Rg3

用於實驗本發明之組合物之效能之人參皂苷Rg3購買自AMBO研究所。 Ginsenoside Rg3, which was used to test the efficacy of the composition of the present invention, was purchased from the AMBO Institute.

[劑型例1及比較劑型例1] [Formulation Example 1 and Comparative Formulation Example 1]

依據下表1之組成，藉由通常之方法而製備營養霜(單位：重量%)。 According to the composition of Table 1 below, a nutrition cream (unit: weight%) was prepared by a usual method.

[試驗例1]氣色改善效果 [Experimental Example 1] Color Improvement Effect

為了評估根據本發明之化妝品組合物之皮膚血液循環促進效果，藉由雷射都普勒血流成像儀(Laser Doppler PeRg3usion Imager；periscan PIM II，Perimed(stochholm，瑞典))而測定皮膚之血液循環程度。雷射都普勒血流成像儀係普遍公知之用於測定於皮膚中之血液循環之儀器，其係目前被使用之儀器，其不僅可測定於皮膚之毛細血管中之血液之速度和量，亦可測定於小動脈和小靜脈中之流動，屬非常敏感之設備。 In order to evaluate the skin blood circulation promoting effect of the cosmetic composition according to the present invention, the blood circulation of the skin was measured by a Laser Doppler PeRg3usion Imager; periscan PIM II, Perimed (stochholm, Sweden) degree. Laser Doppler blood flow imager is a commonly known instrument for measuring blood circulation in the skin. It is a currently used instrument that can not only measure the speed and amount of blood in the capillaries of the skin, It can also measure the flow in small arteries and veins, which is a very sensitive device.

於恆溫恆濕室中，使用香皂洗臉後，放鬆30分鐘，藉由雷射都普勒血流成像儀而測定初始值。首先，針對平時手腳發涼之30名女性，藉由雷射都普勒血流成像儀測定其額頭下端部分之初始血流量。繼而，安排被試者於1週期間內使用上述劑型例1及比較劑型例1，比較所測定之血流量和上述初始測定值，其結果(皮膚血流量變化)顯示於下表2。 After washing the face with soap in a constant temperature and humidity room, relax for 30 minutes, and measure the initial value with a laser Doppler blood flow imager. First, for 30 women who usually have cold hands and feet, the initial blood flow of the lower part of their forehead is measured by a laser Doppler blood flow imager. Next, the subjects were arranged to use the above-mentioned dosage form example 1 and comparative dosage form example 1 within a period of one week, and compared the measured blood flow with the above-mentioned initial measurement value. The results (changes in skin blood flow) are shown in Table 2 below.

自上述表2之結果中，根據本發明之化妝品組合物，其相比於未含有人參皂苷Rg3之比較劑型例1，其顯著增加皮膚血流量，可知藉由此種血液循環之促進而使得氣色得到改善。總之，該結果表示根據本發明之含有人參皂苷Rg3之化妝品組合物，其可貢獻於有效傳遞皮膚之營養成分、抑制和延遲皮膚老化。 From the results in Table 2 above, the cosmetic composition according to the present invention significantly increases skin blood flow compared to Comparative Formulation Example 1 which does not contain ginsenoside Rg3, and it can be seen that the appearance of the complexion is promoted by the promotion of such blood circulation. Improved. In summary, this result indicates that the cosmetic composition containing ginsenoside Rg3 according to the present invention can contribute to effectively transmitting the nutritional ingredients of the skin, suppressing and delaying skin aging.

[試驗例2]膚色改善效果 [Test Example 2] Skin color improvement effect

為了瞭解上述劑型例1及比較劑型例1之膚色改善效果，安排30名被試者分別使用(晚1次/日，為期1週)，藉由Facial Stage DM-3(Moritex，日本)設備而評估膚色改善程度。關於膚色改善率，藉由測定皮膚之亮度及色彩測定值而以其變化值作為膚色改善率，其結果顯示於下表3。亮度及色彩變化值越大，表示其膚色改善程度越高。 In order to understand the skin color improvement effect of the above dosage form example 1 and comparative dosage form example 1, 30 subjects were arranged to use it separately (1 time / day, for 1 week), using Facial Stage DM-3 (Moritex, Japan) equipment and Assess skin tone improvement. Regarding the skin color improvement rate, the skin color improvement rate was measured by measuring the skin brightness and color measurement values, and the results are shown in Table 3 below. The larger the brightness and color change values, the higher the improvement in skin tone.

自上述表3之結果中可知，未含有根據本發明之人參皂苷Rg3之比較劑型例1，其未顯示出顯著之膚色改善效能；相反，含有人參皂苷Rg3並將其作為有效成分之劑型例1，其相比於使用前之膚色，使用後之膚色得到很大改善。 From the results in Table 3 above, it can be seen that Comparative Formulation Example 1 which does not contain ginsenoside Rg3 according to the present invention does not show significant skin tone improving effect; on the contrary, Formulation Example 1 which contains ginsenoside Rg3 as an active ingredient Compared with the skin tone before use, the skin tone after use is greatly improved.

[試驗例3]毛孔收縮效果 [Test Example 3] Pore shrinking effect

1.藉由促進膠原蛋白生物合成之毛孔收縮效果 1. Pore shrinking effect by promoting collagen biosynthesis

藉由與TGF-β之比較而測定根據本發明之人參皂苷Rg3之 膠原蛋白生物合成促進效果。首先，將成纖維細胞(fibroblast)以每孔105個之方式播種(seeding)於24孔(well)，對其進行培養直至生長為90%左右。以無血清杜爾伯科改良伊格爾培養基培養基對其進行24小時之培養後，分別以10ng/ml處理溶解於無血清培養基之本發明之人參皂苷Rg3與TGF-β，於二氧化碳培養基中培養24小時。去除該些上層液，藉由膠原蛋白原型(I)ELISA試劑盒(procollagen type(I)；#MK101，TAKARA(Shiga，日本))而查看膠原蛋白原(procollagen)之增減與否。其結果顯示於表4，關於膠原蛋白之合成性能，其以非處理組100為基準顯示。 The collagen biosynthesis promoting effect of ginsenoside Rg3 according to the present invention was measured by comparison with TGF-β. First, fibroblasts (fibroblast) at 10 5 per well of embodiment seeded (seeding of) in 24-well (Well), its culture was grown until about 90%. After culturing them in serum-free Dulbecco's modified Eagle medium for 24 hours, the ginsenoside Rg3 and TGF-β of the present invention dissolved in serum-free medium were treated at 10 ng / ml, respectively, and cultured in a carbon dioxide medium. 24 hours. The supernatant was removed, and the increase or decrease of the collagen (procollagen) was checked by a collagen prototype (I) ELISA kit (procollagen type (I); # MK101, TAKARA (Shiga, Japan)). The results are shown in Table 4. The synthetic performance of collagen is shown based on the non-treated group 100.

自上述表4之結果中可知，根據本發明之人參皂苷Rg3，其表現出高於作為陽性對照組之TGF-β之程度之優秀性。因而，可知根據本發明之人參皂苷Rg3可增加毛孔周圍之膠原蛋白生成量，從而縮小放大之毛孔。 As can be seen from the results in Table 4 above, the ginsenoside Rg3 according to the present invention exhibits superiority to a degree higher than TGF-β as a positive control group. Therefore, it is known that the ginsenoside Rg3 according to the present invention can increase the amount of collagen produced around the pores, thereby reducing the enlarged pores.

2.毛孔收縮效果 2. Pore shrinking effect

為了瞭解劑型例1及比較劑型例1之毛孔收縮效果，按以下方式進行評估。選定毛孔變大之被試者男女共20名，分為2組每組各10名，各組成員在面部塗抹劑型例1及比較劑型例1之營養霜，每天塗抹連續4週。關於毛孔收縮之效果有關判定，藉由實驗前和實驗4週後之照片，由專家進行肉眼評估。其結果顯 示於下表5(評估等級：0-未見縮小；5-大幅縮小)。 In order to understand the pore shrinkage effect of Formulation Example 1 and Comparative Formulation Example 1, the evaluation was performed in the following manner. Twenty males and females with enlarged pores were selected and divided into two groups of 10 each. The members of each group applied the nutrition cream of Formulation Example 1 and Comparative Formulation Example 1 on the face for 4 weeks each day. Regarding the judgment of the effect of pore shrinkage, the photos were taken by experts before and 4 weeks after the experiment, and evaluated visually by experts. The results show The results are shown in Table 5 below (evaluation level: 0-no shrinkage was seen; 5- drastically shrinkage).

自上述表5之結果中可知，比較劑型例1不具有毛孔收縮效果，而劑型例1則表現出肉眼可看到之顯著之毛孔收縮效果，藉此可知，根據本發明之人參皂苷Rg3，其縮小毛孔大小之效果優秀。 From the results in Table 5 above, it can be seen that Comparative Formulation Example 1 does not have a pore shrinkage effect, while Formulation Example 1 shows a significant pore shrinkage effect that can be seen by the naked eye. From this, it can be known that according to the ginsenoside Rg3 of the present invention, The effect of reducing pore size is excellent.

[試驗例4]皮脂分泌抑制效果 [Test Example 4] Sebum secretion inhibitory effect 藉由1. 5α-還原酶活性抑制之皮膚過分泌抑制效果 Skin over-secretion inhibitory effect by inhibition of 1.5α-reductase activity

為了確認5α-還原酶(5 α-reductase)活性抑制效果，測定於HEK293-5αR2細胞中由[14C]睾酮轉換為[14C]二氫睾酮(DHT：dihydrotestosterone)之比率。將p3 x FLAG-CMV-5αR2轉染至HEK293細胞，以每孔/2.5 x 105細胞之方式放入24孔板並進行培養(Park et al.，2003，JDS.Vol.31，pp.191-98)。次日，更換成加入酶基質與抑制劑之新之培養基。作為培養基之基質而使用0.05μCi[14C]睾酮(Amersham Pharmacia biotech，UK)。 To confirm 5α- reductase (5 α -reductase) activity inhibition effect was measured in the cell HEK293-5αR2 by [14 C] testosterone is converted to [14 C] dihydrotestosterone: ratio (DHT dihydrotestosterone) of. The p3 x FLAG-CMV-5αR2 transfected into HEK293 cells per well /2.5 x 10 5 cells were placed in 24 well plates in such manner and culture (Park et al., 2003, JDS.Vol.31, pp.191 -98). The next day, it was replaced with a new medium with enzyme substrate and inhibitor. As the substrate of the culture medium, 0.05 μCi [ 14 C] testosterone (Amersham Pharmacia biotech, UK) was used.

為了確認5α-還原酶活性抑制程度，加入人參皂苷Rg3並於37℃、5%二氧化碳培養器中培養2小時。此時，將未加入人參皂苷Rg3之組作為陰性對照組，加入非那斯特萊(finasteride)之組作為陽性對照組。繼而，收取培養基，以800μl乙酸乙酯提取類固醇，分離其上部之有機溶劑層進行乾燥，將剩餘之殘留物再 次以50μl乙酸乙酯進行溶解，於矽塑料片矽膠60 F254(Silica plastic sheet kieselgel 60 F254)上，將乙酸乙酯-己烷(1：1)作為溶劑而展開。 In order to confirm the degree of inhibition of 5α-reductase activity, ginsenoside Rg3 was added and cultured in a 5% carbon dioxide incubator at 37 ° C for 2 hours. At this time, the group to which ginsenoside Rg3 was not added was used as a negative control group, and the group to which finasteride was added was used as a positive control group. Then, the medium was collected, and the steroid was extracted with 800 μl of ethyl acetate. The upper organic solvent layer was separated and dried, and the remaining residue was reused. It was dissolved with 50 μl of ethyl acetate at a time, and then developed on a silicone plastic sheet Kieselgel 60 F254 (Silica plastic sheet kieselgel 60 F254) using ethyl acetate-hexane (1: 1) as a solvent for development.

於空氣中乾燥塑料試料後，為了測定同位元素之量而使用沐浴系統，將完成乾燥之塑料片和X射線膠片一同放入沐浴箱(bath cassette)，經過1週後測定殘留於膠片之睾酮和二氫睾酮之同位元素量，依據以下公式1及公式2而分別算出轉化率及抑制率，其結果顯示於下表6。 After drying the plastic sample in the air, use a bathing system to determine the amount of isotopic elements. Put the dried plastic piece together with the X-ray film into the bath cassette. After 1 week, measure the testosterone and The isotopic content of dihydrotestosterone was calculated based on the following formulas 1 and 2, respectively, and the conversion and inhibition rates were calculated. The results are shown in Table 6 below.

[公式1]轉換率(%)=DHT區域之放射能/總放射能×100 [Formula 1] Conversion rate (%) = radioactive energy in the DHT area / total radioactive energy × 100

[公式2]抑制率(%)=(對照組之轉換率-試驗物質之轉換率)/對照組之轉換率×100 [Formula 2] Inhibition rate (%) = (conversion rate of control group-conversion rate of test substance) / conversion rate of control group × 100

自上述表6之結果中，可知人參皂苷Rg3有效抑制5α-還原酶之活性，該5α-還原酶將睾酮轉換為二氫睾酮，使其與細胞質內之水溶體蛋白質相結合，進入核內並活化皮脂腺細胞，促進分化，起到於皮脂腺內過分泌皮脂之作用，藉此，可知人參皂苷Rg3阻斷由睾酮轉換為二氫睾酮，其表現出比用於抑制5α-還原酶之活性之非那斯特萊更加優秀之抑制效果。因而可知，人參皂苷Rg3藉由有效抑制5α-還原酶之活性而抑制皮脂之過分泌。 From the results in Table 6 above, it can be seen that ginsenoside Rg3 effectively inhibits the activity of 5α-reductase, which converts testosterone to dihydrotestosterone, combines it with water-soluble proteins in the cytoplasm, enters the nucleus, and Activates sebaceous gland cells, promotes differentiation, and acts as a supersecretory sebum in the sebaceous glands. Based on this, it can be seen that ginsenoside Rg3 blocks the conversion from testosterone to dihydrotestosterone, which shows a higher effect than that used to inhibit the activity of 5α-reductase. Nastelier has more excellent suppression effect. Therefore, it is known that ginsenoside Rg3 inhibits the secretion of sebum by effectively inhibiting the activity of 5α-reductase.

2.皮脂分泌抑制效果 2. Sebum secretion inhibitory effect

為了瞭解上述劑型例1及比較劑型例1之皮脂分泌抑制效果，按以下方式進行評估。選定認為皮脂分泌多之男女性被試者共30名，於面部皮膚之指定部位塗抹劑型例1及比較劑型例1之營養霜，每天塗抹連續4週。關於皮脂減少之效果有關判定，藉由皮脂量測定儀器(Sebumeter SM810，C+K Electronic Co.，德國)而分別測定經過2週及4週後之平均皮脂減少率(%)，其結果顯示於下表7。 In order to understand the effect of suppressing sebum secretion of the above-mentioned formulation example 1 and comparative formulation example 1, the evaluation was performed in the following manner. A total of 30 male and female subjects who were considered to have high sebum secretion were selected, and the nutrient creams of Formulation Example 1 and Comparative Formulation Example 1 were applied to a specified part of the facial skin, and applied for 4 weeks each day. Regarding the determination of the effect of sebum reduction, the average sebum reduction rate (%) after 2 weeks and 4 weeks was measured by a sebum volume measuring instrument (Sebumeter SM810, C + K Electronic Co., Germany), and the results are shown in Table 7 below.

自上述表7之結果中可知，藉由含有本發明之人參皂苷Rg3而將其作為有效成分之劑型例1，相比於未含有其之比較劑型例1，其可有效抑制過量分泌之皮脂。 From the results in Table 7 above, it can be seen that the dosage form example 1 containing the ginsenoside Rg3 of the present invention as an active ingredient can effectively suppress the excessive secretion of sebum compared to the comparative dosage form example 1 not containing it.

[劑型例2及比較劑型例2~3] [Formulation example 2 and comparative formulation examples 2 to 3]

根據下表8所示之成分及含量(重量%)而製備劑型例2及比較劑型例2~3。具體而言，劑型例2中配合有人參皂苷Rg3，比較劑型例2中完全不含有用於改善痤瘡皮膚之有效成分，比較劑型例3係作為抗菌力有關基準之標準物質，含有多用作痤瘡治療劑之紅黴素(erythromycin)。 Formulation example 2 and comparative formulation examples 2 to 3 were prepared based on the components and contents (% by weight) shown in Table 8 below. Specifically, Ginsenoside Rg3 is blended in Formulation Example 2, and Comparative Formulation Example 2 does not contain effective ingredients for improving acne skin at all. Comparative Formulation Example 3 is a standard substance related to antibacterial efficacy, and it is mostly used for acne treatment. Erythromycin.

劑型例2及比較劑型例2~3之製備方法如下。完全溶解下表8之A之成分，於獨立之溶解槽中完全溶解B之成分，將B添加至A並實施混合溶解化。其中，按照表8所示之配比加入C 之成分，均勻混合後過濾而製備本組合物。 The preparation methods of Formulation Example 2 and Comparative Formulation Examples 2 to 3 are as follows. The components of A in Table 8 below were completely dissolved, and the components of B were completely dissolved in a separate dissolution tank. B was added to A and mixed and dissolved. Among them, C is added according to the ratio shown in Table 8. The ingredients are uniformly mixed and then filtered to prepare the composition.

[試驗例5]對痤瘡之抗菌力試驗 [Test Example 5] Antibacterial effect test on acne

藉由以上述劑型例2及比較劑型例2~3之組成而製備之各化妝品組合物，對作為痤瘡病源菌株之痤瘡丙酸桿菌(Propionibacterium acnes，P.acnes)(ATCC 6919：培養基-BHI培養基(broth))進行抗菌力試驗。 By using each of the cosmetic compositions prepared with the composition of the above formulation example 2 and comparative formulation examples 2 to 3, Propionibacterium acnes (P. acnes) (ATCC 6919: culture medium-BHI medium), which is an acne-producing strain, is prepared. (broth)) An antibacterial test was performed.

痤瘡有關抗菌力試驗方法如下所示。 The acne-related antibacterial test method is shown below.

(1)試驗菌液準備 (1) Preparation of test bacteria liquid

痤瘡丙酸桿菌使用接種於BHI培養基而進行厭氧培養之培養液。 Propionibacterium acnes uses a culture medium inoculated in BHI medium and anaerobic cultured.

(2)稀釋溶液準備 (2) Preparation of diluted solution

將0.15ml上述試驗菌液加入15ml之BHI培養基(pH 6.8)或LB培養基(pH 4.5)，均勻混合，作為稀釋溶液使用。 0.15 ml of the test bacterial solution was added to 15 ml of BHI medium (pH 6.8) or LB medium (pH 4.5), mixed uniformly, and used as a diluted solution.

(3)試料準備 (3) Sample preparation

將以劑型例2及比較劑型例2~3製備之化妝品組合物原液直接作為試料而使用。 The cosmetic composition stock solutions prepared in Formulation Example 2 and Comparative Formulation Examples 2 to 3 were directly used as samples.

(4)抗菌力試驗 (4) Antibacterial test

1)向96孔之細胞培養管(96 well plate)1號行，按照起始濃度，放入試料，作為稀釋溶液，以200μl為單位加入總量。 1) In a 96-well cell culture tube (96 well plate), line 1, place the sample at the initial concentration, and use it as a diluted solution. Add 200 μl as the unit.

2)均勻混合1號行之混合液後，取100μl並放入2號行，均勻混合，再次提取100μl並放入3號行，實施兩倍稀釋(double dilution)。 2) After uniformly mixing the mixed solution in line 1 and taking 100 μl and placing it in line 2, uniformly mixing, extracting 100 μl again and putting it in line 3, and carry out double dilution.

3)於32℃中實施24小時及48小時靜置培養後，藉由懸浮程度而判斷菌之增殖與否，將沒有菌增殖之最小濃度作為最低抑制濃度(MIC，Minimum Inhibitory Concentration)值。若混合液不透明而難以判斷菌之增殖與否，則藉由顯微鏡觀察而進行確認。 3) After performing incubation for 24 hours and 48 hours at 32 ° C, the proliferation of bacteria is judged by the degree of suspension, and the minimum concentration without bacteria proliferation is taken as the minimum inhibitory concentration (MIC, Minimum Inhibitory Concentration) value. If the mixed solution is opaque and it is difficult to judge the proliferation of the bacteria, it is confirmed by microscopic observation.

痤瘡有關抗菌力試驗結果顯示於表9。最低抑制濃度換算成含於劑型之有效成分之濃度而進行標記。 The results of the acne-related antibacterial test are shown in Table 9. The minimum inhibitory concentration is marked as the concentration of the active ingredient contained in the dosage form.

自上述表9之結果中可知，最低抑制濃度之ppm濃度越小，其於痤瘡有關抗菌力方面可作為更有效之物質，劑型例2相比於使用作為公知之痤瘡治療劑之紅黴素之比較劑型例3，其ppm濃度明顯小，藉此可知，含有人參皂苷Rg3之組合物對試驗菌具有顯著優秀之抗菌力。 From the results in Table 9 above, it can be seen that the smaller the ppm concentration of the lowest inhibitory concentration, the more effective it is in acne-related antibacterial properties. Formulation Example 2 is compared with the use of erythromycin, which is a known acne treatment agent. In Comparative Formulation Example 3, the ppm concentration was remarkably small. From this, it can be seen that the composition containing ginsenoside Rg3 has a significantly excellent antibacterial effect on the test bacteria.

[試驗例6]脂質合成(Lipogenesis)抑制試驗 [Test Example 6] Lipogenesis inhibition test

將老鼠之成纖維細胞系(fibroblast cell line)之3T3-L1細胞，以1×105細胞/孔之方式，附著於盛放含有10%之胎牛血清 (fetal bovine serum，FBS)之杜爾伯科改良伊格爾培養基(Dulbecos modified eagles medium，GIBCO BRL，Life Technologes公司)培養基之6孔培養板(culture plate)。經過2天後，重新更換成新之杜爾伯科改良伊格爾培養基(含有10%之FBS)培養基，繼續培養2天。繼而，針對完成培養之上述細胞，以含有1μg/ml胰島素(insulin)、0.5mM IBMX及0.25μM***(dexamethasone)之杜爾伯科改良伊格爾培養基(含有10%之FBS)實施分化誘導，處理人參皂苷Rg3及咖啡因50μM，經過處理2天後，再次更換為包含胰島素之杜爾伯科改良伊格爾培養基，繼續培養5天。經過5天後，再次更換為正常培養基(杜爾伯科改良伊格爾培養基，含有10%之FBS)，一面觀察一面培養，直至上述細胞於形態方面變化為脂肪細胞。 3T3-L1 cells of mouse fibroblast cell line were attached to Dürr containing 10% fetal bovine serum (FBS) at 1 × 10 5 cells / well. A 6-well culture plate of Dulbecos modified eagles medium (GIBCO BRL, Life Technologes). After 2 days, the medium was replaced with a new Dulberco modified Igel medium (containing 10% FBS), and the culture was continued for 2 days. Then, the above-mentioned cells that were cultured were differentiated in a Dulberco's modified Igel medium (containing 10% FBS) containing 1 μg / ml insulin, 0.5 mM IBMX, and 0.25 μM dexamethasone. Induction and treatment of ginsenoside Rg3 and caffeine 50 μM, after 2 days of treatment, it was replaced again with Dulboco's modified Eagle's medium containing insulin, and culture was continued for 5 days. After 5 days, the medium was replaced with normal medium (Durberco's modified Eagle's medium containing 10% FBS), and cultured while observing, until the above cells changed into morphologically fat cells.

為了評估人參皂苷Rg3之對脂肪細胞內脂肪堆積之抑制效能，藉由上述過程中完成分化之3T3-L1脂肪細胞，實施蘇丹III染色(S4136，sigma-aldrich)。將脂肪細胞於磷酸鹽緩衝液內以4%之多聚甲醛(pH 7.2)於常溫下固定後，以磷酸鹽緩衝液(PBS，phosphate buffered saline)水洗，再以蘇丹III染色後，拍攝圖片並進行肉眼比較。對照組係僅使用未添加試驗物質或比較物質之培養基，作為其他比較組，則處理咖啡因50μM。關於脂肪堆積抑制程度，藉由將染色程度劃分為+++、++、+、-而賦予等級，此時，越接近+++，其意味著染色程度越高。其結果顯示於下表10。 In order to evaluate the inhibitory effect of ginsenoside Rg3 on fat accumulation in adipocytes, Sudan III staining (S4136, sigma-aldrich) was performed on the differentiated 3T3-L1 adipocytes in the above process. Adipocytes were fixed in phosphate buffered saline with 4% paraformaldehyde (pH 7.2) at room temperature, washed with phosphate buffered saline (PBS), and stained with Sudan III. Take pictures and Make a visual comparison. The control group only used the medium without adding the test substance or the comparative substance. As the other comparison group, the caffeine was treated at 50 μM. Regarding the degree of fat accumulation inhibition, the degree of staining is classified by dividing the degree of staining into +++, ++, +, and-. In this case, the closer to +++, the higher the degree of staining. The results are shown in Table 10 below.

自上述表10之結果中可知，根據本發明之人參皂苷Rg3，不僅脂肪細胞內堆積之脂肪之量少，而且相比於公知之作為脂質合成抑制物質之咖啡因，其具有更優秀之脂質合成抑制效果。因而，藉由抑制脂質合成而減少皮脂，從而可抑制痤瘡之生成。 From the results in Table 10 above, it can be seen that according to the ginsenoside Rg3 of the present invention, not only the amount of fat accumulated in fat cells is small, but also it has better lipid synthesis than caffeine, which is known as a lipid synthesis inhibitor. Inhibitory effect. Therefore, by inhibiting lipid synthesis and reducing sebum, the generation of acne can be suppressed.

[試驗例7]痤瘡改善與皮脂分泌減少以及刺激有無相關試驗 [Experimental Example 7] Tests on improvement of acne, reduction of sebum secretion, and presence or absence of stimulation

將患有痤瘡之30名被試者以每組10名之方式共分為3組，安排各組被試者分別使用以上述劑型例2及比較劑型例2~3製備之化妝品組合物。痤瘡改善尺度設置為1分至5分，1分為“不是”、3分為“一般”、5分為“非常是”。實驗結果以下表11之10名被試者之平均分數進行標記。 Thirty subjects with acne were divided into three groups in a manner of 10 in each group, and the subjects in each group were arranged to use the cosmetic composition prepared in the above dosage form example 2 and comparative dosage forms examples 2 to 3 respectively. The acne improvement scale is set from 1 to 5 points, 1 being "not", 3 being "fair", and 5 being "very yes". The experimental results are marked with the average scores of the 10 participants in Table 11 below.

痤瘡消失時期係以所讀取之痤瘡消失之相應日數為基準，痤瘡再發有無係以一個月之後之結果為基準。皮脂分泌減少尺度設置為1分至5分，1分為“不是”、3分為“一般”、5分為“非常是”。實驗結果以下表11之10名被試者之平均分數進行標記。皮膚刺激之有無以(表現出刺激反應之人數)/(被試者總人數)表示。 The period of acne disappearance is based on the number of days when the acne disappearance is read. The occurrence of acne recurrence is based on the result after one month. Sebum secretion reduction scale is set to 1 to 5 points, 1 is "not", 3 is "fair", and 5 is "very yes". The experimental results are marked with the average scores of the 10 participants in Table 11 below. The presence or absence of skin irritation is indicated by (the number of people who show a stimulus response) / (the total number of subjects).

自上述表11之結果中可知，劑型例2相比於比較劑型例 2，其痤瘡未再發，整體上，其對痤瘡改善方面具有優秀之效果。另外，於含有抗菌力標準物質之比較劑型例3中，雖然表現出痤瘡改善效果，然而於使用過程中，其表現出強皮膚刺激性，其不適合長期使用，而根據本發明之組合物，則無刺激性，表現出可長期使用性。 As can be seen from the results in Table 11 above, Formulation Example 2 is compared to Comparative Formulation Example 2. Its acne has not recurred. On the whole, it has an excellent effect on improving acne. In addition, in Comparative Formulation Example 3 containing an antibacterial standard substance, although it shows an acne-improving effect, it shows strong skin irritation during use, which is not suitable for long-term use. However, according to the composition of the present invention, Non-irritating, showing long-term use.

[劑型例3及比較劑型例4] [Formulation example 3 and comparative formulation example 4]

以下表12之組成製備洗髮乳。具體而言，將表面活性劑和雙硬脂酸乙二醇酯(ethylene glycol distearate)添加至純淨水，加熱至80℃並均勻溶解後，攪拌並逐步降溫至40℃，向上述混合物中加入根據本發明之有效成分和防腐劑、黏度調節劑、香料、毛髮調理劑後，攪拌並冷卻至室溫，完成製備。 The composition of Table 12 below was used to prepare a shampoo. Specifically, add surfactant and ethylene glycol distearate to purified water, heat to 80 ° C and dissolve uniformly, stir and gradually reduce the temperature to 40 ° C, add to the above mixture according to After the effective ingredients and preservatives, viscosity modifiers, perfumes, and hair conditioners of the present invention, they are stirred and cooled to room temperature to complete the preparation.

[試驗例8]抗頭屑效果試驗 [Test Example 8] Anti-dandruff effect test

選定頭屑較多之19~35歲之男性24名，以每組12名之方式共分成2組，安排各組分別使用劑型例3及比較劑型例4之洗髮乳，連續使用1個月後，測定其頭屑減少率。 Twenty-nine men aged 19-35 years with more dandruff were selected and divided into two groups with 12 in each group. Each group was arranged to use shampoo in dosage form 3 and comparative dosage form 4 for 1 month. Then, the dandruff reduction rate was measured.

在開始試驗之前，使用一般洗髮乳洗髮，採集洗髮後2天內堆積之頭屑，測量其頭屑之重量，再分別使用劑型例3及比較劑型例4之洗髮乳每隔兩天洗髮1次，採集完成試驗後2天內堆積之頭屑，測量其頭屑之重量，兩者進行比較。此時，藉由真空吸入裝置自頭皮直接採集堆積之頭屑，依據以下公式3而求出頭屑減少率，其結果顯示於下表13。 Before starting the test, use a normal shampoo to shampoo, collect the dandruff accumulated within 2 days after shampooing, measure the weight of the dandruff, and then use the shampoos of Formulation Example 3 and Comparative Formulation Example 4 every two times. Shampoo once a day, collect dandruff accumulated within 2 days after the test is completed, measure the weight of dandruff, and compare the two. At this time, the accumulated dandruff was directly collected from the scalp by the vacuum suction device, and the dandruff reduction rate was obtained according to the following formula 3. The results are shown in Table 13 below.

[公式3]頭屑減少率(%)={試驗開始前頭屑重量(mg)-一個月後頭屑重量(mg)}/試驗開始前頭屑重量(mg)×100 [Formula 3] Dandruff reduction rate (%) = {dandruff weight before the test (mg)-dandruff weight after one month (mg)} / dandruff weight before the test (mg) × 100

自上述表13之結果中可知，含有人參皂苷Rg3之劑型例3表現出優秀之抗頭屑效果。 As can be seen from the results in Table 13 above, Formulation Example 3 containing ginsenoside Rg3 exhibited an excellent antidandruff effect.

[試驗例9]頭皮瘙癢症防止效果試驗 [Test Example 9] Test for Preventing Scalp Pruritus

選定具有較嚴重之頭皮瘙癢症之25歲~45歲之男女性共24名，每組以12名共分成2組，安排使用各劑型例3及比較劑型例4之洗髮乳，每3天1次連續使用2週，藉由以下評估基準而評估頭皮瘙癢症防止效果，其結果顯示於表14。 A total of 24 men and women aged 25 to 45 with severe pruritus of the scalp were selected, and each group was divided into 2 groups with 12 members. The shampoos of Example 3 and Comparative Example 4 were arranged for use every 3 days. The results were shown in Table 14 after using it continuously for 2 weeks, and evaluated the scalp pruritus prevention effect according to the following evaluation criteria.

[評估標準] [Evaluation Criteria]

非常優秀-5分 Very good -5 points

優秀-4分 Excellent -4 points

一般-3分 Average -3 points

不合格-2分 Unqualified-2 points

非常不合格-1分 Very unqualified-1 point

自上述表14之結果中可知，含有人參皂苷Rg3之劑型例3於防止頭皮瘙癢方面表現出更優秀之效果。 As can be seen from the results in Table 14 above, Formulation Example 3 containing ginsenoside Rg3 exhibited a more excellent effect in preventing itching of the scalp.

[試驗例10]鉀離子通道活性增加效果評估 [Experimental Example 10] Evaluation of the effect of increasing potassium ion channel activity

作為脫髮治療劑之米諾地爾(Minoxidil)被認為係潛在之粒線體鉀離子通道開放劑(KATP channel opener)，其係用於雄激素性脫髮之代表性藥物。為了評估此種米諾地爾之機制，使用如下試驗方法，即，於構成頭皮之真皮之成纖維細胞中處理用於堵塞粒線體鉀離子通道之甲苯磺丁脲(SIGMA AlDRICH，T0891)而抑制細胞增殖，再次打開鉀離子通道並恢復細胞增殖。 Minoxidil as a hair loss treatment agent is considered to be a potential mitochondrial potassium ion channel opener (K ATP channel opener), which is a representative drug for androgenic alopecia. In order to evaluate the mechanism of such minoxidil, a test method was used in which tolbutamide (SIGMA AlDRICH, T0891) for blocking mitochondrial potassium ion channels was treated in fibroblasts constituting the dermis of the scalp, and Inhibit cell proliferation, reopen potassium channels and restore cell proliferation.

為了評估本組合物之作為粒線體鉀離子通道開放劑之功能，於本發明中使用成纖維細胞株之NIH3T3(Mouse embryonic fibroblast cell line)細胞株。本細胞株係將分離自NIH瑞士小鼠胚胎(Swiss mouse embryo)之成纖維細胞株以3T3協議進行天然永生之細胞株。上述細胞株於含10%之FBS之杜爾伯科改良伊格爾培養基(Gibco BRL，Gaithersburg，MD，美國)中，於保持5%二氧化碳、37℃之培養器中培養24小時。將NIH3T3放入96孔板，於37℃培養器中培養24小時後處理2.5mM甲苯磺丁脲，經過10分鐘後，將作為陽性對照組之10μM米諾地爾和人參皂苷Rg3分別以2.5ppm、5ppm及10ppm之濃度處理，藥物處理經過48小時後，使用WST-1試劑盒(羅氏)而測定細胞增殖能力。其結果顯示於下表15。 In order to evaluate the function of the composition as a mitochondrial potassium ion channel opener, the fibroblast cell line NIH3T3 (Mouse embryonic fibroblast cell line) cell line. This cell line will be a natural immortal cell line derived from the fibroblast cell line of the NIH Swiss mouse embryo under the 3T3 protocol. The above cell lines were cultured in a Dulberco modified Igel medium (Gibco BRL, Gaithersburg, MD, USA) containing 10% FBS in an incubator maintained at 5% carbon dioxide at 37 ° C for 24 hours. Put NIH3T3 into a 96-well plate, incubate in a 37 ° C incubator for 24 hours, and then treat 2.5 mM tolbutamide. After 10 minutes, 10 μM minoxidil and ginsenoside Rg3 were used as positive control groups at 2.5 ppm. Treatment at concentrations of 5 ppm and 10 ppm. After 48 hours of drug treatment, the cell proliferation ability was measured using a WST-1 kit (Roche). The results are shown in Table 15 below.

於上述表15之結果中可知，若處理人參皂苷Rg3，則恢復成纖維細胞之增殖，隨著所處理之人參皂苷Rg3之濃度增加，其細胞增殖能力亦增加，若以10ppm處理人參皂苷Rg3，則細胞增殖恢復程度達到處理米諾地爾之情況之程度。 From the results in Table 15 above, it can be seen that if ginsenoside Rg3 is treated, the proliferation of fibroblasts is restored. As the concentration of the treated ginsenoside Rg3 increases, its cell proliferation ability also increases. If ginsenoside Rg3 is treated at 10 ppm, Then, the degree of cell proliferation recovery is as high as that in the case of treating minoxidil.

[試驗例11]人參皂苷Rg3之黑色素生成促進效果試驗 [Experimental Example 11] A melanin production promoting effect test of ginsenoside Rg3

向RPMI培養基添加5%之胎牛血清、100IU之青黴素G及0.2μM之TPA，將黑色素細胞(melan-a)向24孔板(24-well microtiter plate)按照50,000細胞/孔進行分散。次日，向分散之細胞以最終濃度10ppm或50ppm處理作為試驗物質之人參皂苷 Rg3，作為陰性對照組處理0.1%之DMSO、作為陽性對照組處理100μM IBMX後，於37℃之溫度下培養3天。完成培養後，使用磷酸鹽緩衝液(PBS)清洗孔，按照每次100μl之量放入1N氫氧化鈉(NaOH)，溶解細胞內之黑色素。關於溶解之黑色素之吸光度，藉由平板培養測定儀器(microplate reader)於405nm中進行測定(Synergy2，BioTek(VT，美國))。人參皂苷Rg3之黑色素生成促進效果與對照組進行比較之結果顯示於下表16。 5% fetal calf serum, 100 IU of penicillin G, and 0.2 μM of TPA were added to the RPMI medium, and melan-a was dispersed into a 24-well microtiter plate at 50,000 cells / well. The next day, the dispersed cells were treated with ginsenoside as a test substance at a final concentration of 10 ppm or 50 ppm. Rg3 was treated with 0.1% DMSO as a negative control group and 100 μM IBMX as a positive control group, and then cultured at 37 ° C. for 3 days. After the culture was completed, the wells were washed with phosphate buffered saline (PBS), and 1N sodium hydroxide (NaOH) was added in an amount of 100 μl each time to dissolve the melanin in the cells. The absorbance of the dissolved melanin was measured at 405 nm using a microplate reader (Synergy2, BioTek (VT, USA)). The melanin production promotion effect of ginsenoside Rg3 is compared with the control group, and the results are shown in Table 16 below.

自上述表16之結果中可知，人參皂苷Rg3促進黑素細胞之黑色素合成，增加黑色素生成量，從而表現出優秀之黑色素生成促進效果。 From the results in Table 16 above, it can be seen that ginsenoside Rg3 promotes melanin synthesis in melanocytes and increases the amount of melanin production, thereby exhibiting an excellent melanin production promotion effect.

[試驗例12]於人參皂苷Rg3之黑素細胞中之小眼畸形相關轉錄因子(MITF)及酪氨酸酶(tyrosinase)表現促進效果 [Experimental Example 12] Effect of promoting expression of small eye malformation-related transcription factor (MITF) and tyrosinase in melanocytes of ginsenoside Rg3

藉由501mel之細胞株而向6孔板(6-well microtiter plate)按照500,000細胞/孔而進行分散，針對各孔，作為陰性對照組處理DMSO 0.1%、作為陽性對照組處理IBMX 100μM、以及作為試驗組以10ppm處理人參皂苷Rg3，於37℃溫度下培養24小時、48小時、72小時後獲取蛋白質。對於如此獲取之蛋白質，藉由小眼畸形相關轉錄因子(MITF)及酪氨酸酶抗體而實施免疫印跡。蛋白質提取和免疫印跡通常使用本領域技術人員採用之標 準方法。完成免疫印跡後，其結果以陰性對照組之100為準進行比較，其結果顯示於下表17。 501mel cell lines were dispersed into 6-well microtiter plates at 500,000 cells / well. For each well, 0.1% DMSO was treated as a negative control group, IBMX 100 μM was treated as a positive control group, and The test group treated ginsenoside Rg3 at 10 ppm and cultured at 37 ° C for 24 hours, 48 hours, and 72 hours to obtain protein. The protein thus obtained was subjected to immunoblotting with small eye malformation-related transcription factor (MITF) and tyrosinase antibody. Protein extraction and western blotting are usually performed using standards adopted by those skilled in the art. Quasi-method. After the immunoblot was completed, the results were compared with 100 of the negative control group, and the results are shown in Table 17 below.

自上述表17之結果中可知，人參皂苷Rg3可提高黑素細胞中之小眼畸形相關轉錄因子(MITF)和酪氨酸酶蛋白質表現。 From the results in Table 17 above, it is known that ginsenoside Rg3 can improve the expression of small eye malformation-related transcription factor (MITF) and tyrosinase protein in melanocytes.

[試驗例13]人參皂苷Rg3之抗菌力評估 [Experimental Example 13] Evaluation of Antibacterial Effect of Ginsenoside Rg3

為了評估人參皂苷Rg3之抗菌力而實施抗菌實驗。具體方法如下所述。 In order to evaluate the antibacterial effect of ginsenoside Rg3, an antibacterial experiment was performed. The specific method is described below.

用於試驗之金黃色葡萄球菌(Staphylococcus aureus)、大腸桿菌(Escherichia coli)、綠膿桿菌(Pseudomonas aeruginosa)菌株於胰酪腖大豆培養基(Tryptic Soy Broth)中進行培養，白色念珠菌(Candida albicans)、黑曲黴菌(Aspergillus niger)菌株於沙保氏葡萄糖培養液(Sabouraud Dextrose Broth)中進行培養。將培養液1/100(白色念珠菌菌株為1/10)稀釋至各培養基之稀釋液作為試驗菌液而使用。黑曲黴菌將以按照2×108cfu/ml而製備之孢子懸浮液作為試驗菌液而使用。 Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa strains used for the experiments were cultured in Tryptic Soy Broth, and Candida albicans 1. Aspergillus niger strains were cultured in Sabouraud Dextrose Broth. The culture solution was diluted 1/100 (1/10 of Candida albicans strain) to each culture medium and used as a test bacterial solution. Aspergillus niger was used as a test bacterial solution, a spore suspension prepared at 2 × 10 8 cfu / ml.

向各15ml培養基添加0.15ml之上述試驗菌液，均勻混合並作為稀釋溶液而使用。 0.15 ml of the above-mentioned test bacteria solution was added to each 15 ml of the culture medium, and they were uniformly mixed and used as a diluted solution.

向96孔板(96 well plate)1號行中以每次16μl放入人參皂苷Rg310ppm，各放入稀釋溶液184μl。向剩餘之孔，各放入稀釋 溶液100μl。均勻混合1號行之混合液後，提取100μl並放入2號行，均勻混合後，再次提取100μl並放入3號行，藉由此方式而實施2倍稀釋。 In row 96 of a 96-well plate (96 well plate), ginsenoside Rg 310 ppm was placed at 16 μl each time, and 184 μl of a diluted solution was placed in each. Dilute each of the remaining holes 100 μl of solution. After uniformly mixing the mixed solution of line 1 and extracting 100 μl and placing it in line 2, after uniformly mixing, extracting 100 μl again and placing it in line 3, in this way, a 2-fold dilution was performed.

金黃色葡萄球菌(Staphylococcus aureus)、大腸桿菌(Escherichia coli)、綠膿桿菌(Pseudomonas aeruginosa)於32℃之恆溫槽中進行培養，白色念珠菌(Candida albicans)、黑曲黴菌(Aspergillus niger)於25℃之恆溫槽中進行培養。 Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were cultured in a thermostatic bath at 32 ° C. Candida albicans and Aspergillus niger were cultured at 25 ° C. Cultivate in a constant temperature bath at ℃.

48小時後，藉由顯微鏡確認菌增殖與否和懸浮度，確定最低抑制濃度(MIC)值並將其結果顯示於下表18。 After 48 hours, the presence or absence of bacterial proliferation and the degree of suspension were confirmed by a microscope, and the minimum inhibitory concentration (MIC) value was determined, and the results are shown in Table 18 below.

自上述表18之結果中可知，人參皂苷Rg3對各種菌株表現出抗菌力，藉此可預測，人參皂苷Rg3可於組合物內作為天然防腐劑或抗菌劑而發揮作用。 From the results in Table 18 above, it can be seen that ginsenoside Rg3 exhibits antibacterial power against various strains, and thus it can be predicted that ginsenoside Rg3 can function as a natural preservative or antibacterial agent in the composition.

Claims (6)

一種改善氣色及膚色的方法，包括：在一個體的皮膚上施用含有人參皂苷Rg3為唯一有效成分的皮膚外用劑組合物，以增加皮膚血流量。A method for improving complexion and complexion, comprising: applying a skin external preparation composition containing ginsenoside Rg3 as the sole active ingredient to the skin of an individual to increase skin blood flow. 一種促進毛髮生長的方法，包括：在一個體的頭皮上施用含有人參皂苷Rg3為唯一有效成分的皮膚外用劑組合物，以打開細胞的粒線體鉀離子通道(KATPchannel)。A method for promoting hair growth, comprising: applying a skin external preparation composition containing ginsenoside Rg3 as a sole active ingredient on a scalp of a body to open a cell's mitochondrial potassium ion channel (K ATP channel). 一種使用人參皂苷Rg3來製備氣色及膚色改善用組成物的用途，其中人參皂苷Rg3為其中唯一有效成分且用來增加皮膚血流量。A use of ginsenoside Rg3 to prepare a composition for improving color and skin tone, wherein ginsenoside Rg3 is the only active ingredient therein and is used to increase skin blood flow. 一種使用人參皂苷Rg3來製備抑制皮膚脂質分泌用組成物的用途，其中人參皂苷Rg3為其中唯一有效成分且用來抑制細胞中5α-還原酶的活性。A use of ginsenoside Rg3 to prepare a composition for inhibiting skin lipid secretion, wherein ginsenoside Rg3 is the only active ingredient therein and is used to inhibit the activity of 5α-reductase in cells. 一種使用人參皂苷Rg3來製備改善痤瘡用組成物的用途，其中人參皂苷Rg3為其中唯一有效成分且用來抑制痤瘡丙酸桿菌感染痤瘡。A use of ginsenoside Rg3 to prepare a composition for improving acne, wherein ginsenoside Rg3 is the only active ingredient therein and is used to inhibit P. acnes infection of acne. 一種使用人參皂苷Rg3來製備毛髮生長促進用組成物的用途，其中人參皂苷Rg3為其中唯一有效成分且用來促進細胞的粒線體鉀離子通道打開。A use of ginsenoside Rg3 to prepare a composition for promoting hair growth, wherein ginsenoside Rg3 is the only active ingredient therein and is used to promote the opening of mitochondrial potassium ion channels of cells.
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