TW201831166A - Composition comprising ginseng flower extracts with enhanced ginsenoside content - Google Patents
Composition comprising ginseng flower extracts with enhanced ginsenoside content Download PDFInfo
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- TW201831166A TW201831166A TW107102194A TW107102194A TW201831166A TW 201831166 A TW201831166 A TW 201831166A TW 107102194 A TW107102194 A TW 107102194A TW 107102194 A TW107102194 A TW 107102194A TW 201831166 A TW201831166 A TW 201831166A
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- ginseng flower
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Abstract
Description
本發明揭露一組成物，其包括作為有效成分之人參皂苷含量提升之人參花萃取物且其係用於提升皮膚屏障功能；及一方法，其係用於製備該組成物。The present invention discloses a composition comprising ginseng flower extract having an increased ginsenoside content as an active ingredient and which is used for enhancing skin barrier function; and a method for preparing the composition.
人參(Panax ginseng C.A. Meyer)係刺五加(Eleutherococcus senticosus)科人參屬之植物。其在韓國、中國及日本等被用作為草藥超過2,000年。其實證上係用於預防疾病及延長壽命。人參之效能迄今為止已知包括對中樞神經系統之效果、抗致癌(anticarcinogenic)活性、抗癌(anticance)活性、免疫功能調節、抗糖尿病活性、肝功能改善效果、心血管疾病改善、抗動脈粥狀硬化(antiatherogenic)活性、血壓控制活性、改善更年期疾病、對骨質疏鬆症之效果、抗壓力活性、抗疲勞活性、抗氧化活性及抗衰老效果。 Panax ginseng CA Meyer is a plant of the genus Echinacea of the family Eleutherococcus senticosus. It has been used as a herbal medicine in Korea, China and Japan for more than 2,000 years. In fact, it is used to prevent diseases and prolong life. The efficacy of ginseng has been known to date to include effects on the central nervous system, anticarcinogenic activity, anticancer activity, immune function regulation, antidiabetic activity, liver function improvement, cardiovascular disease improvement, anti-atherosclerosis Antiatherogenic activity, blood pressure control activity, improvement of menopausal disease, effect on osteoporosis, anti-stress activity, anti-fatigue activity, antioxidant activity and anti-aging effect.
人參皂苷(Ginsenoside)(人參之代表性生理有效成分)均勻地分佈在人參之地上及地下部分中。然而，人參皂苷之含量及組成物已知會依據人參之部分(諸如根、葉、果等)而不同。Ginsenoside (a representative physiologically active ingredient of ginseng) is evenly distributed on the ground and underground parts of ginseng. However, the content and composition of ginsenosides are known to vary depending on the parts of ginseng (such as roots, leaves, fruits, etc.).
近來，隨著消費者對天然化妝品之興趣增加及推出許多使用藥草之化妝品，對於人參作為在皮膚化妝品具有重要功效之植物材料已進行研究。然而，大部分之研究只針對人參根部萃取物或人參根部之人參皂苷及人參多醣體進行，沒有對果實等之人參各部分之成分功效進行研究。此外，長期以來一直人參花之價值都未知而大部分人參花被丟掉，不如人參種子。最近人參花(具有鮮明之清新氣味)作為芳香植物之價值已被注意，而開始發展能表現出人參花香氣之薰香組成物。然而，仍然未知人參花對皮膚之功效(諸如提升皮膚屏障)而對此缺乏研究。Recently, as consumers' interest in natural cosmetics has increased and many cosmetics using herbs have been introduced, ginseng has been studied as a plant material having an important effect on skin cosmetics. However, most of the studies were conducted only on ginseng root extracts or ginseng saponins and ginseng polysaccharides in ginseng roots, and there was no study on the efficacy of the components of ginseng and other parts of the fruit. In addition, the value of ginseng flowers has long been unknown and most ginseng flowers have been lost, not as good as ginseng seeds. Recently, ginseng flowers (with a distinct fresh scent) have been noted as the value of aromatic plants, and the development of aroma components that can express ginseng flower aroma has begun. However, there is still no known research on the efficacy of ginseng flowers on the skin, such as lifting the skin barrier.
與此同時，由人參獲得萃取物之傳統方法存在著問題，其不適合用於降低細胞毒性同時保持人參藥理成分。At the same time, the conventional method of obtaining extracts from ginseng has problems, which are not suitable for reducing cytotoxicity while maintaining ginseng pharmacological components.
[技術問題][technical problem]
在一態樣中，本發明目標之一在於提供具有提升之人參皂苷Re含量之人參花萃取物之新穎用途。In one aspect, one of the objects of the present invention is to provide a novel use of a ginseng flower extract having an elevated ginsenoside Re content.
在另一態樣中，本發明目標之一在於提供一組成物，其包括作為有效成分之人參花萃取物且其具有提升之皮膚屏障功能。In another aspect, one of the objects of the present invention is to provide a composition comprising ginseng flower extract as an active ingredient and having an enhanced skin barrier function.
在另一態樣中，本發明目標之一在於提供一用於製備該組成物之方法。 [問題解決方案]In another aspect, one of the objects of the present invention is to provide a method for preparing the composition. [problem solution]
在一態樣中，本發明提供一用於提升皮膚屏障功能之組成物，其包括作為有效成分之人參花萃取物，其包括以100重量份作為萃取原料之乾燥人參花為基礎至少2.30重量份之人參皂苷Re。In one aspect, the present invention provides a composition for enhancing a skin barrier function, comprising as an active ingredient a ginseng flower extract comprising at least 2.30 parts by weight based on 100 parts by weight of dry ginseng flower as an extraction raw material. Ginsenoside Re.
在另一態樣中，本發明提供一用於製備該組成物之方法，其包括以下步驟：將水、一有機溶劑或一混合水與有機溶劑之溶劑加入至人參花，然後反復加壓減壓萃取製程以執行萃取。 [發明有益效果]In another aspect, the present invention provides a method for preparing the composition comprising the steps of: adding water, an organic solvent or a solvent of a mixed water and an organic solvent to a ginseng flower, and then repeatedly reducing the pressure. The extraction process is performed to perform the extraction. [Advantageous Effects of Invention]
在一態樣中，本發明之包括作為有效成分之人參花萃取物，其包括以100重量份作為萃取原料之乾燥人參花為基礎之至少2.30重量份之人參皂苷Re之該組成物會增加抗微生物胜肽基因(antimicrobial peptide genes)hBD2、hBD3及牛皮癬素(psoriasin)之表現，其可作為皮膚表面之抗微生物屏障因此提升皮膚屏障功能。In one aspect, the present invention comprises, as an active ingredient, a ginseng flower extract comprising at least 2.30 parts by weight of ginsenoside Re based on 100 parts by weight of dried ginseng flower as an extraction raw material, the composition increases resistance The performance of antimicrobial peptides hBD2, hBD3 and psoriasin can act as an antimicrobial barrier to the skin surface and thus enhance the skin barrier function.
下文將詳細描述本發明。The invention will be described in detail below.
在一態樣中，本發明提供一用於提升皮膚屏障功能之組成物，其包括作為有效成分之人參花萃取物，以100重量份作為萃取原料之乾燥人參花為基礎其包括至少2.30重量份之人參皂苷Re。In one aspect, the present invention provides a composition for enhancing a skin barrier function, comprising as an active ingredient a ginseng flower extract comprising at least 2.30 parts by weight based on 100 parts by weight of dry ginseng flower as an extraction raw material. Ginsenoside Re.
本發明之人參花(Panax ginseng C. A. Meyer)屬於人參之地上部分。相較於人參根其含有較多之人參皂苷，而其類型與人參根類似。未收集種子之人參花會被去除以促進人參根之生長。The ginseng flower ( Panax ginseng CA Meyer) of the present invention belongs to the aerial part of ginseng. Compared with ginseng root, it contains more ginsenosides, and its type is similar to ginseng root. Ginseng flowers that have not been collected will be removed to promote the growth of ginseng roots.
本發明之人參皂苷係一種皂苷(saponin) (一種醣苷(glycoside))，其源自植物之根、枝幹、葉、皮部、種子等。人參皂苷有各種種類，包含人參皂苷 Rb1、Rb2、Rc、Rd、Re、Rg1等。人參皂苷之種類及含量隨著植物種類、栽培條件、加工條件、萃取方法、人參部位等而有不同。根據結構，人參皂苷可被分類成PPT(Re、Rg1)及PPD(Rb1、Rb2、Rc、Rd)。Rb1、Rb2、Rc、Rd、Re及Rg1為水溶性極性物質，其具有天然人參之組成物，亦即其未進行去除糖之變性作用以製備出紅參。The ginseng saponin of the present invention is a saponin (a glycoside) derived from the roots, branches, leaves, skins, seeds and the like of plants. There are various types of ginsenosides, including ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1 and the like. The type and content of ginsenoside vary depending on the plant species, cultivation conditions, processing conditions, extraction methods, and ginseng parts. According to the structure, ginsenosides can be classified into PPT (Re, Rg1) and PPD (Rb1, Rb2, Rc, Rd). Rb1, Rb2, Rc, Rd, Re and Rg1 are water-soluble polar substances having a composition of natural ginseng, that is, they are not subjected to sugar-reducing denaturation to prepare red ginseng.
本發明之人參花萃取物可能包括以100重量份作為萃取原料之乾燥人參花為基礎之以下量之人參皂苷Re：至少2.30重量份、至少2.32重量份、至少2.34重量份、至少2.36重量份、至少2.38重量份、至少2.40重量份、至少2.42重量份、至少2.44重量份、至少2.46重量份、至少2.48重量份、至少2.49重量份、至少2.50重量份、至少2.52重量份、至少2.54重量份、至少2.56重量份、至少2.58重量份、至少2.60重量份、至少2.62重量份、至少2.63重量份、至少2.64重量份、至少2.66重量份、至少2.68重量份、至少2.70重量份、至少2.72重量份、至少2.74重量份、至少2.76重量份、至少2.78重量份、至少2.8重量份、至少2.82重量份、至少2.84重量份、至少2.86重量份、至少2.88重量份、至少2.90重量份、至少2.92重量份、至少2.94重量份、至少2.96重量份、2.98重量份或至少3.0重量份。The ginseng flower extract of the present invention may comprise the following amount of ginsenoside Re based on 100 parts by weight of dry ginseng flower as an extraction raw material: at least 2.30 parts by weight, at least 2.32 parts by weight, at least 2.34 parts by weight, at least 2.36 parts by weight, At least 2.38 parts by weight, at least 2.40 parts by weight, at least 2.42 parts by weight, at least 2.44 parts by weight, at least 2.46 parts by weight, at least 2.48 parts by weight, at least 2.49 parts by weight, at least 2.50 parts by weight, at least 2.52 parts by weight, at least 2.54 parts by weight, At least 2.56 parts by weight, at least 2.58 parts by weight, at least 2.60 parts by weight, at least 2.62 parts by weight, at least 2.63 parts by weight, at least 2.64 parts by weight, at least 2.66 parts by weight, at least 2.68 parts by weight, at least 2.70 parts by weight, at least 2.72 parts by weight, At least 2.74 parts by weight, at least 2.76 parts by weight, at least 2.78 parts by weight, at least 2.8 parts by weight, at least 2.82 parts by weight, at least 2.84 parts by weight, at least 2.86 parts by weight, at least 2.88 parts by weight, at least 2.90 parts by weight, at least 2.92 parts by weight, At least 2.94 parts by weight, at least 2.96 parts by weight, 2.98 parts by weight or at least 3.0 parts by weight.
本發明之人參花萃取物可能包括以100重量份作為萃取原料之乾燥人參花為基礎之以下者：至少0.25重量份、至少0.26重量份、至少0.27重量份、至少0.28重量份、至少0.29重量份或至少0.30重量份之人參皂苷Rb1，至少0.20重量份、至少0.21重量份、至少0.22重量份、至少0.23重量份、至少0.24重量份或至少0.25重量份之人參皂苷Rb2，或至少0.20重量份、至少0.22重量份、至少0.24重量份、至少0.26重量份、至少0.28重量份、至少0.30重量份、至少0.31重量份、至少0.32重量份、至少0.33重量份或至少0.34重量份之人參皂苷Rc。而且，本發明之人參花萃取物可能包括以100重量份作為萃取原料之乾燥人參花為基礎之以下者：至少0.60重量份、至少0.62重量份、至少0.64重量份、至少0.66重量份、至少0.68重量份、至少0.70重量份、至少0.72重量份、至少0.74重量份、至少0.76重量份、至少0.78重量份、至少0.80重量份、至少0.81重量份、至少0.82重量份、至少0.83重量份或至少0.84重量份之人參皂苷(ginsenoside) Rd，或至少0.25重量份、至少0.26重量份、至少0.27重量份、至少0.28重量份、至少0.29重量份、至少0.30重量份或至少0.31重量份之人參皂苷Rg1。在本發明之一具體實施例中，發現於24小時、水溫30°C、加壓萃取壓力80 Mpa及減壓萃取壓力一大氣壓之條件(實施例1至3)下以該加壓減壓萃取製程製備之該人參花萃取物具有較高產量且以100 g人參花粉末為基礎之人參皂苷Re之絕對量增加且相較於以一般萃取製程(比較實施例1至3)製備之人參花萃取物含有較多之人參皂苷(測試實施例1至3及圖1)。The ginseng flower extract of the present invention may comprise, based on 100 parts by weight of dry ginseng flower as an extraction raw material, at least 0.25 parts by weight, at least 0.26 parts by weight, at least 0.27 parts by weight, at least 0.28 parts by weight, at least 0.29 parts by weight. Or at least 0.30 parts by weight of ginsenoside Rb1, at least 0.20 parts by weight, at least 0.21 parts by weight, at least 0.22 parts by weight, at least 0.23 parts by weight, at least 0.24 parts by weight or at least 0.25 parts by weight of ginsenoside Rb2, or at least 0.20 parts by weight, At least 0.22 parts by weight, at least 0.24 parts by weight, at least 0.26 parts by weight, at least 0.28 parts by weight, at least 0.30 parts by weight, at least 0.31 parts by weight, at least 0.32 parts by weight, at least 0.33 parts by weight or at least 0.34 parts by weight of ginsenoside Rc. Moreover, the ginseng flower extract of the present invention may comprise at least 0.60 parts by weight, at least 0.62 parts by weight, at least 0.64 parts by weight, at least 0.66 parts by weight, at least 0.68, based on 100 parts by weight of dry ginseng flowers as an extraction raw material. Parts by weight, at least 0.70 parts by weight, at least 0.72 parts by weight, at least 0.74 parts by weight, at least 0.76 parts by weight, at least 0.78 parts by weight, at least 0.80 parts by weight, at least 0.81 parts by weight, at least 0.82 parts by weight, at least 0.83 parts by weight or at least 0.84 Parts by weight of ginsenoside Rd, or at least 0.25 parts by weight, at least 0.26 parts by weight, at least 0.27 parts by weight, at least 0.28 parts by weight, at least 0.29 parts by weight, at least 0.30 parts by weight or at least 0.31 parts by weight of ginsenoside Rg1. In one embodiment of the present invention, the pressure is reduced under conditions of 24 hours, a water temperature of 30 ° C, a pressure extraction pressure of 80 MPa, and a vacuum extraction pressure of one atmosphere (Examples 1 to 3). The ginseng flower extract prepared by the extraction process has a higher yield and the absolute amount of ginsenoside Re based on 100 g of ginseng flower powder is increased and compared with the ginseng flower prepared by the general extraction process (Comparative Examples 1 to 3). The extract contained more ginsenosides (Test Examples 1 to 3 and Figure 1).
本發明之皮膚屏障(角質層)係由角質細胞和細胞間脂質所組成。其保護皮膚不受外界刺激、防止皮膚水氣蒸發及扮演皮膚健康之重要角色。The skin barrier (stratum corneum) of the present invention consists of keratinocytes and intercellular lipids. It protects the skin from external stimuli, prevents skin moisture from evaporating and plays an important role in skin health.
本發明之抗微生物胜肽係一與皮膚表面之抗微生物屏障有關之抗微生物胜肽。當抗微生物胜肽基因之表現量增加，該皮膚屏障功能可被進一步強化。本發明之抗微生物胜肽基因可能為一或多之基因，其係選自於由人類β防禦素2(hBD2)、人類β防禦素3(hBD3)及牛皮癬素(psoriasin)所組成之群組。hBD2及hBD3屬於哺乳動物之防禦素家族。β-防禦素具有一抗微生物胜肽功能因此顯示出對大量革蘭氏陰性(gram-negative)或革蘭氏陽性(gram-positive)菌、真菌等之抗性。牛皮癬素(psoriasin)(也被稱為S100A7)屬於S100蛋白質家族。其係由皮膚上皮細胞分泌並沉積以幫助形成抗微生物屏障。在本發明之一具體實施例中，發現當以在24小時、水溫30°C、加壓萃取壓力80 Mpa及減壓萃取壓力一大氣壓之條件(實施例3)製備之該人參花萃取物處理角質細胞，hBD2、hBD3及牛皮癬素(psoriasin)基因之表現量會高於其在未處理組(控制組)、熱水萃取 (比較實施例4)及在大氣壓以一般萃取製程製備之萃取物之表現量(比較實施例3)。因此，發現本發明之包括至少一具體量之人參皂苷Re之人參花萃取物藉由增加抗微生物胜肽之表現來提升皮膚屏障功能(測試實施例4及圖2)。The antimicrobial peptide of the present invention is an antimicrobial peptide associated with the antimicrobial barrier of the skin surface. The skin barrier function can be further enhanced when the amount of expression of the antimicrobial peptide gene is increased. The antimicrobial peptide gene of the present invention may be one or more genes selected from the group consisting of human beta defensin 2 (hBD2), human beta defensin 3 (hBD3), and psoriasin. . hBD2 and hBD3 belong to the mammalian family of defensins. The beta-defensin has an anti-microbial peptide function and thus exhibits resistance to a large number of gram-negative or gram-positive bacteria, fungi, and the like. Psoriasin (also known as S100A7) belongs to the S100 protein family. It is secreted and deposited by skin epithelial cells to help form an antimicrobial barrier. In a specific embodiment of the present invention, the ginseng flower extract prepared under the conditions of 24 hours, a water temperature of 30 ° C, a pressure extraction pressure of 80 MPa, and a vacuum extraction pressure of one atmosphere (Example 3) was found. Treatment of keratinocytes, hBD2, hBD3 and psoriasin genes will be more abundant than in the untreated group (control group), hot water extraction (Comparative Example 4) and extracts prepared at atmospheric pressure in a general extraction process. The amount of performance (Comparative Example 3). Thus, it was found that the ginseng flower extract of the present invention comprising at least one specific amount of ginsenoside Re enhances the skin barrier function by increasing the performance of the antimicrobial peptide (Test Example 4 and Figure 2).
任何方法可能被使用以製備本發明之人參花萃取物，只要其能夠製備包括以100重量份作為萃取原料之乾燥人參花為基礎之至少2.30重量份之人參皂苷Re之人參花萃取物。而且，該方法可能使用一加壓減壓萃取製程。該加壓減壓萃取製程可能為一進行加壓製程或一減壓製程用以在萃取製程中改變壓力。具體地，本發明之人參花萃取物可能藉由交替進行加壓製程及減壓製程之壓力循環萃取製程來製備，且其可能連續進行。該壓力循環萃取製程可能藉由將水、有機溶劑或其混合溶劑加入至人參花及於規律間隔反復進行加壓製程及減壓製程來進行。更具體地，該加壓減壓萃取製程可能反復進行18小時或其以上、19小時或其以上、20小時或其以上、21小時或其以上、22小時或其以上、23小時或其以上、或24小時或其以上及30小時或其以下、29小時或其以下、28小時或其以下、27小時或其以下、26小時或其以下、25小時或其以下、或24小時或其以下。該加壓減壓萃取製程之萃取溫度可能為25°C或更高、26°C或更高、27°C或更高、28°C或更高、29°C或更高、或30°C或更高及35°C或更低、34°C或更低、33°C或更低、32°C或更低、31°C或更低、或30°C或更低。在該加壓萃取製程期間中該壓力可能為60 MPa或更高、65MPa或更高、70 MPa或更高、75 MPa或更高、或80 MPa或更高及100 MPa或更低、95 MPa或更低、90 MPa或更低、85 MPa或更低、或80 MPa或更低。而且，在該加壓萃取製程期間中該壓力可能為一大氣壓力。Any method may be used to prepare the ginseng flower extract of the present invention as long as it can prepare a ginseng flower extract comprising at least 2.30 parts by weight of ginsenoside Re based on 100 parts by weight of dry ginseng flower as an extraction raw material. Moreover, the process may use a pressurized vacuum extraction process. The pressurized decompression extraction process may be a pressurization process or a depressurization process to change the pressure during the extraction process. Specifically, the ginseng flower extract of the present invention may be prepared by a pressure cycle extraction process in which a pressurization process and a depressurization process are alternately performed, and it may be continuously performed. The pressure cycle extraction process may be carried out by adding water, an organic solvent or a mixed solvent thereof to the ginseng flower and repeating the pressurization process and the pressure reduction process at regular intervals. More specifically, the pressurized vacuum extraction process may be repeated for 18 hours or more, 19 hours or more, 20 hours or more, 21 hours or more, 22 hours or more, 23 hours or more, Or 24 hours or more and 30 hours or less, 29 hours or less, 28 hours or less, 27 hours or less, 26 hours or less, 25 hours or less, or 24 hours or less. The extraction temperature of the pressurized vacuum extraction process may be 25 ° C or higher, 26 ° C or higher, 27 ° C or higher, 28 ° C or higher, 29 ° C or higher, or 30 ° C or higher and 35 ° C or lower, 34 ° C or lower, 33 ° C or lower, 32 ° C or lower, 31 ° C or lower, or 30 ° C or lower. The pressure may be 60 MPa or higher, 65 MPa or higher, 70 MPa or higher, 75 MPa or higher, or 80 MPa or higher, and 100 MPa or lower, 95 MPa during the pressurized extraction process. Or lower, 90 MPa or lower, 85 MPa or lower, or 80 MPa or lower. Moreover, the pressure may be an atmospheric pressure during the pressurized extraction process.
在本發明之一具體實施例中，發現該用於製備具有最佳產量及人參皂苷Re之最高絕對量之人參花萃取物之加壓減壓萃取製程最佳條件係24小時處理時間、水溫30°C及加壓萃取壓力80 MPa(測試實施例2)。In a specific embodiment of the present invention, it is found that the optimum conditions for the preparation of the ginseng flower extract having the highest yield and the highest absolute amount of ginsenoside Re are the 24-hour treatment time and the water temperature. 30 ° C and a pressure extraction pressure of 80 MPa (Test Example 2).
本發明之組成物可能包括以該組成物總重為基礎之以下量之人參花萃取物：0.0001 wt%至99 wt%，具體為0.1 wt%至99 wt%，更具體為1 wt%至50 wt%。The composition of the present invention may comprise the following amount of ginseng flower extract based on the total weight of the composition: 0.0001 wt% to 99 wt%, specifically 0.1 wt% to 99 wt%, more specifically 1 wt% to 50. Wt%.
在此態樣中，該組成物可能為一藥學組成物、一化妝品組成物或一食品組成物。In this aspect, the composition may be a pharmaceutical composition, a cosmetic composition or a food composition.
該藥學組成物可被配製成固體、半固體或液態之口服或腸胃外劑型，其係藉由將無機或有機載體加入至該作為有效成分之組成物。The pharmaceutical composition can be formulated into a solid, semi-solid or liquid oral or parenteral dosage form by adding an inorganic or organic carrier to the composition as an active ingredient.
口服劑型之實施例包含錠劑(tablets)、丸劑(pills)、顆粒劑(granules)、軟或硬膠囊(capsules)、散劑(powders)、細顆粒(fine granules)、粉劑(dusts)、乳劑(emulsions)、糖漿劑(syrups)、微粒劑(pellets)等。腸胃外給藥劑型之實例包含注射液劑(injections)、滴劑(drops)、軟膏劑(ointments)、洗劑(lotions)、噴霧劑(sprays)、懸浮液(suspensions)、乳劑(emulsions)、栓劑(suppositories)等。本發明之有效成分可容易地根據傳統方法配製。表面活性劑、賦形劑、著色劑、香料、防腐劑、穩定劑、緩衝劑、懸浮劑及其他常用佐劑可被適當地使用。Examples of oral dosage forms include tablets, pills, granules, soft or hard capsules, powders, fine granules, dusts, emulsions ( Emulsions), syrups, pellets, and the like. Examples of parenteral dosage forms include injections, drops, ointments, lotions, sprays, suspensions, emulsions, Suppositories and the like. The active ingredient of the present invention can be easily formulated according to a conventional method. Surfactants, excipients, colorants, perfumes, preservatives, stabilizers, buffers, suspending agents, and other conventional adjuvants can be suitably used.
本發明之藥學組成物對於提升皮膚屏障有優異之效果，因此可被有用地應用在治療及預防皮膚屏障損傷造成之皮膚病。皮膚屏障損傷造成之皮膚病包含但不限於異位性皮膚炎(atopic dermatitis)、乾皮症(xeroderma)、牛皮癬(psoriasis)、魚鱗癬(ichthyosis)、痤瘡(acne)等。The pharmaceutical composition of the present invention has an excellent effect on lifting the skin barrier, and thus can be usefully used for treating and preventing skin diseases caused by skin barrier damage. Skin diseases caused by skin barrier damage include, but are not limited to, atopic dermatitis, xeroderma, psoriasis, ichthyosis, acne, and the like.
該藥學組成物可經由口、腸胃外、直腸、局部、透皮、靜脈內、肌內、腹膜內、皮下等施予。The pharmaceutical composition can be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, and the like.
此外，該有效成分之劑量將隨著受治療對象之年齡性別及體重、疾病或病況之嚴重程度、施予途徑及處方者判斷而改變。基於此等因素決定劑量係在本領域技術人員之知識範圍內。典型劑量為0.001 mg/kg/日至2000 mg/kg/日，更具體為0.5 mg/kg/日至1500 mg/kg/日。In addition, the dose of the active ingredient will vary depending on the age, sex and weight of the subject, the severity of the disease or condition, the route of administration, and the judgment of the prescriber. Determining the dosage based on such factors is within the knowledge of those skilled in the art. Typical dosages are from 0.001 mg/kg/day to 2000 mg/kg/day, more specifically from 0.5 mg/kg/day to 1500 mg/kg/day.
可能以任何適合用於局部施用之配方提供該化妝品組成物。 舉例而言，可能以下列者提供：一液體，將一油相分散在一水相中獲得之乳劑，將一水相分散在一油相中獲得之乳劑，一懸浮液，一固體，一凝膠，一粉末，一糊劑，一泡沫或一氣霧劑組成物。此等配方之組成物可能根據該領域中之傳統方法製備。The cosmetic composition may be provided in any formulation suitable for topical application. For example, it may be provided by a liquid, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, and a solid. Glue, a powder, a paste, a foam or an aerosol composition. The compositions of such formulations may be prepared according to conventional methods in the art.
除了上述材料外，該化妝品組成物可能含有能提供協同效應而不會負面影響主要效果之其他成分。本發明之化妝品組成物可能包括一材料，其選自於由以下所組成之群組：維生素，高分子量肽，高分子量多醣及神經鞘脂(sphingolipids)。而且，本發明之化妝品組成物可能包括濕潤劑、潤滑劑、表面活性劑、紫外線吸收劑、防腐劑、殺菌劑、抗氧化劑、pH調節劑、有機及無機色素、香料、冷卻劑或止汗劑。此等成分之量可容易地由本領域技術人員在對本發明目的及效果沒有負面影響之範圍內決定。該成分之含量以該組成物之總重為基礎可能為0.001至5 wt%，具體為0.01至3 wt%。In addition to the above materials, the cosmetic composition may contain other ingredients that provide a synergistic effect without adversely affecting the primary effect. The cosmetic composition of the present invention may comprise a material selected from the group consisting of vitamins, high molecular weight peptides, high molecular weight polysaccharides, and sphingolipids. Moreover, the cosmetic composition of the present invention may include a wetting agent, a lubricant, a surfactant, a UV absorber, a preservative, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a coolant or an antiperspirant. . The amounts of such ingredients can be readily determined by those skilled in the art without departing from the scope and effect of the invention. The content of the component may be 0.001 to 5 wt%, specifically 0.01 to 3 wt%, based on the total weight of the composition.
在此態樣中，該藥學組成物或化妝品組成物可能為一皮膚外用組成物，舉例而言，一霜劑或軟膏劑。In this aspect, the pharmaceutical composition or cosmetic composition may be a skin external composition, for example, a cream or ointment.
該食品組成物可能為一健康食品組成物。其可能為一需要發酵之發酵食品組成物，諸如茶葉、乳製品、泡菜及釀造食品。該食品組成物之配方未受特別限制，但其可能為錠劑(tablets)、丸劑(pills)、軟或硬膠囊(capsules)、顆粒劑(granules)、飲料(drinks)、焦糖(caramels)、能量棒(diet bars)、茶包(tea bags)等。除了有效成分外，各配方之食品組成物可能進一步包括常用於相關領域之成分。該成分可能由該領域技術人士無困難地根據所欲之配方或目的選擇或添加。其他成分之添加可能會產生協同效應。該有效成分劑量之決定係在本領域技術人士之知識範圍內。該有效成分之劑量可能隨著各種因素改變，其包含受試者之年齡及健康條件、併發症等。The food composition may be a healthy food composition. It may be a fermented food composition that requires fermentation, such as tea, dairy, kimchi, and brewed food. The formulation of the food composition is not particularly limited, but it may be tablets, pills, soft or hard capsules, granules, drinks, caramels. , diet bars, tea bags, etc. In addition to the active ingredients, the food compositions of the respective formulations may further include ingredients commonly used in related fields. This ingredient may be selected or added by a person skilled in the art without difficulty depending on the desired formula or purpose. The addition of other ingredients may have a synergistic effect. The determination of the active ingredient dose is within the knowledge of those skilled in the art. The dosage of the active ingredient may vary with various factors including the subject's age and health conditions, complications, and the like.
在另一態樣中，本發明可關於一用於預防、改善或治療皮膚屏障受損所致之皮膚病之方法，其包括將一有效量之人參花萃取物給予需要預防、改善或治療皮膚屏障受損所致之皮膚病之受試者，以100重量份作為萃取原料之乾燥人參花為基礎其包括至少2.30重量份之人參皂苷Re。在本發明一態樣中，該方法之給藥可根據此處所述之給藥方法及給藥劑量進行。In another aspect, the invention may be directed to a method for preventing, ameliorating or treating a skin disorder caused by impaired skin barrier comprising administering an effective amount of a ginseng flower extract to a skin in need of prevention, amelioration or treatment. The subject of the skin disease caused by the damage of the barrier comprises at least 2.30 parts by weight of ginsenoside Re based on 100 parts by weight of dry ginseng flower as an extraction raw material. In one aspect of the invention, administration of the method can be carried out according to the methods of administration and dosages administered herein.
在另一態樣中，本發明可能關於人參花萃取物用於製備能提升皮膚屏障功能之化妝品組成物之用途，以100重量份作為萃取原料之乾燥人參花為基礎其包括至少2.30重量份之人參皂苷Re。In another aspect, the present invention may be directed to the use of a ginseng flower extract for the preparation of a cosmetic composition capable of enhancing the barrier function of the skin, comprising at least 2.30 parts by weight based on 100 parts by weight of dry ginseng flower as an extraction raw material. Ginsenoside Re.
在另一態樣中，本發明可能關於人參花萃取物用於製備能提升皮膚屏障功能之食品組成物之用途，以100重量份作為萃取原料之乾燥人參花為基礎其包括至少 2.30重量份之人參皂苷Re。In another aspect, the present invention may be directed to the use of a ginseng flower extract for the preparation of a food composition capable of enhancing skin barrier function, comprising at least 2.30 parts by weight based on 100 parts by weight of dry ginseng flower as an extraction material. Ginsenoside Re.
在另一態樣中，本發明可能關於能提升皮膚屏障功能之人參花萃取物，以100重量份作為萃取原料之乾燥人參花為基礎其包括至少 2.30重量份之人參皂苷Re。In another aspect, the present invention may be directed to a ginseng flower extract capable of enhancing skin barrier function comprising at least 2.30 parts by weight of ginsenoside Re based on 100 parts by weight of dry ginseng flower as an extraction raw material.
在另一態樣中，本發明提供一用於製備該組成物之方法，其包括以下步驟：將作為主要溶劑之水或醇加入至人參花然後反復一加壓減壓萃取製程以執行主要萃取。而且，該製備方法進一步包括以下步驟：以含氯之有機溶劑分餾該生成之主要萃取物，去除該含氯之有機溶劑層，然後以作為輔助溶劑之醇萃取該水層。當該主要溶劑為醇，該方法可能包括在以一含氯之有機溶劑分餾前將該主要萃取物溶於水中。In another aspect, the present invention provides a method for preparing the composition comprising the steps of: adding water or an alcohol as a main solvent to a ginseng flower and repeating a pressurized decompression extraction process to perform main extraction . Moreover, the preparation method further comprises the steps of fractionating the main extract formed with a chlorine-containing organic solvent, removing the chlorine-containing organic solvent layer, and then extracting the aqueous layer with an alcohol as an auxiliary solvent. When the primary solvent is an alcohol, the process may include dissolving the primary extract in water prior to fractional distillation with a chlorine-containing organic solvent.
該醇可能為一具有1至5個碳原子之低級醇，其包含甲醇、乙醇、丁醇、丙醇及異丙醇等。而且，其可能為一有機溶劑，舉例而言，如乙酸乙酯之極性溶劑及如之如己烷或二氯甲烷之非極性溶劑或其混合物。具體地，該醇可能為10至90% (v/v)乙醇，更具體為80% (v/v)乙醇。本發明之主要溶劑可能為水或醇，而該輔助溶劑可能為醇。具體地，該主要溶劑可能為水，而該輔助溶劑可能為乙醇。The alcohol may be a lower alcohol having 1 to 5 carbon atoms, which comprises methanol, ethanol, butanol, propanol, isopropanol and the like. Moreover, it may be an organic solvent, for example, a polar solvent such as ethyl acetate and a non-polar solvent such as hexane or dichloromethane or a mixture thereof. Specifically, the alcohol may be from 10 to 90% (v/v) ethanol, more specifically 80% (v/v) ethanol. The primary solvent of the present invention may be water or an alcohol, and the auxiliary solvent may be an alcohol. Specifically, the primary solvent may be water and the auxiliary solvent may be ethanol.
該含氯之有機溶劑(其係一含氯之有機溶劑)可能為一或多選自於由二氯甲烷、氯仿、氯甲烷及四氯化碳所組成之群組。具體地，其可能為二氯甲烷。The chlorine-containing organic solvent, which is a chlorine-containing organic solvent, may be one or more selected from the group consisting of dichloromethane, chloroform, methyl chloride, and carbon tetrachloride. Specifically, it may be dichloromethane.
該主要溶劑、輔助溶劑及含氯之有機溶劑可能彼此相同或不同。The main solvent, the auxiliary solvent, and the chlorine-containing organic solvent may be the same or different from each other.
當該加壓減壓萃取製程係如上述方式進行，可最小化該人參花之熱變性從而最小化人參皂苷之變性。因此，相較於不加壓減壓而使用習知簡單萃取裝置萃取人參花之方式，該萃取產量可被提高10%或更多。When the pressurized vacuum extraction process is carried out as described above, the thermal denaturation of the ginseng flower can be minimized to minimize the denaturation of ginsenoside. Therefore, the extraction yield can be increased by 10% or more as compared with the method of extracting ginseng flowers by a conventional simple extraction device without pressure reduction.
此外，使用含氯之有機溶劑(如二氯甲烷)能夠去除葉綠素，而以有機溶劑(如乙醇)二次萃取該水層可最大化人參皂苷之含量。In addition, the use of a chlorine-containing organic solvent such as methylene chloride can remove chlorophyll, and the secondary extraction of the aqueous layer with an organic solvent such as ethanol can maximize the content of ginsenoside.
以下將參考以下實施例及測試實施例來描述本發明。然而，以下實施例及測試實施例僅用於說明目的以便於理解本發明，而本發明之範圍並未受限於此。The invention will be described below with reference to the following examples and test examples. However, the following examples and test examples are for illustrative purposes only, and the scope of the present invention is not limited thereto.
[比較實施例] 使用一般萃取製程製備人參花萃取物[Comparative Example] Preparation of Ginseng Flower Extract by General Extraction Process
[比較實施例1及2] 使用一般萃取製程製備人參花萃取物粉末 以純水清洗由韓國尚州及安東收集之人參花，以熱空氣乾燥或自然乾燥，然後粉末化以獲得細顆粒之人參花粉末。然後在低溫下儲存該粉末。然後，將10L水加入至0.5kg人參花粉末，於30°C萃取該混合物。該所得之萃取物過濾及濃縮至固體含量50%。進行二氯甲烷(MC)分餾用以自該所得之濃縮物去除葉綠素。然後取該水層，向其加入乙醇直至成為80％乙醇。浸漬12小時以去除糖後，過濾溶液丟棄沉澱物。然後，濃縮濾液直到去除乙醇以製得125g人參花萃取物粉末(比較實施例1及2)。[Comparative Examples 1 and 2] The ginseng flower extract powder was prepared by a general extraction process, and the ginseng flower collected from Shangzhou and Anton, Korea, was washed with pure water, dried by hot air or naturally dried, and then powdered to obtain fine granules of ginseng. Flower powder. The powder is then stored at low temperature. Then, 10 L of water was added to 0.5 kg of ginseng flower powder, and the mixture was extracted at 30 °C. The resulting extract was filtered and concentrated to a solids content of 50%. Dichloromethane (MC) fractionation is performed to remove chlorophyll from the resulting concentrate. The aqueous layer was then taken and ethanol was added thereto until it became 80% ethanol. After immersing for 12 hours to remove the sugar, the filtrate was filtered to discard the precipitate. Then, the filtrate was concentrated until ethanol was removed to obtain 125 g of ginseng flower extract powder (Comparative Examples 1 and 2).
[比較實施例3] 根據一般萃取製程製備人參花萃取物以分析人參皂苷及確定抗微生物胜肽之表現 用與比較實施例1及2相同方法在24小時、水溫30°C及大氣壓力之條件下處理人參花(紫莖變種(violet-stem variant))以製備125g人參花萃取物(比較實施例3)。[Comparative Example 3] Preparation of ginseng flower extract according to a general extraction process to analyze ginsenosides and determination of antimicrobial peptide performance The same method as in Comparative Examples 1 and 2 was carried out at 24 hours, water temperature of 30 ° C and atmospheric pressure. Ginseng flowers (violet-stem variant) were treated under conditions to prepare 125 g of ginseng flower extract (Comparative Example 3).
[比較實施例4] 根據熱水萃取法製備人參花萃取物以確定抗微生物胜肽之表現 將10L水加入至0.5kg人參花(紫莖變種)粉末，於95°C萃取該混合物。過濾該所得之萃取物及濃縮至固體含量50%。然後，進行以上與比較實施例1及2之相同方法(如二氯甲烷分餾、添加乙醇、過濾及濃縮)以製備125g人參花萃取物(比較實施例4)。[Comparative Example 4] Preparation of ginseng flower extract according to hot water extraction method to determine the performance of the antimicrobial peptide 10 L of water was added to 0.5 kg of ginseng flower (purple stem) powder, and the mixture was extracted at 95 °C. The resulting extract was filtered and concentrated to a solid content of 50%. Then, the same procedures as in Comparative Examples 1 and 2 (e.g., dichloromethane fractionation, ethanol addition, filtration, and concentration) were carried out to prepare 125 g of ginseng flower extract (Comparative Example 4).
[實施例] 根據加壓減壓萃取製程製備人參花萃取物[Examples] Preparation of Ginseng Flower Extract by Pressurized Vacuum Extraction Process
[實施例1及2] 根據加壓減壓萃取製程製備人參花萃取物粉末 以純水清洗由韓國尚州及安東收集之人參花，以熱空氣乾燥或自然乾燥，然後粉末化以獲得細顆粒之人參花粉末。然後在低溫下儲存該粉末。然後，將10L水加入至0.5kg人參花粉末，然後以反復加壓減壓萃取製程進行萃取24小時，各萃取使用壓力循環萃取裝置進行15分鐘。該加壓萃取壓力為80Mpa及該減壓萃取壓力係大氣壓力。該萃取溫度被設定為30°C。過濾及濃縮藉以上製程獲得之該人參花萃取物至固體含量50%。自該所得之濃縮物進行二氯甲烷(MC)分餾以去除葉綠體。然後取該水層，向其加入乙醇直到成為80%乙醇。浸漬12小時以去除糖後，過濾溶液丟棄沉澱物。然後，濃縮濾液直到去除乙醇以製得125g人參花萃取物粉末(實施例1及2)[Examples 1 and 2] Preparation of Ginseng Flower Extract Powder by Pressurized Vacuum Decompression Extraction Process Ginseng flowers collected from Shangzhou and Anton, Korea, were washed with pure water, dried by hot air or naturally dried, and then powdered to obtain fine particles. Ginseng flower powder. The powder is then stored at low temperature. Then, 10 L of water was added to 0.5 kg of ginseng flower powder, and extraction was carried out for 24 hours under repeated pressure and reduced pressure extraction processes, and each extraction was carried out for 15 minutes using a pressure cycle extraction apparatus. The pressurized extraction pressure was 80 MPa and the reduced pressure extraction pressure was atmospheric pressure. The extraction temperature was set to 30 °C. The ginseng flower extract obtained by the above process was filtered and concentrated to a solid content of 50%. The resulting concentrate was subjected to fractional distillation in dichloromethane (MC) to remove chloroplasts. The aqueous layer was then taken and ethanol was added thereto until it became 80% ethanol. After immersing for 12 hours to remove the sugar, the filtrate was filtered to discard the precipitate. Then, the filtrate was concentrated until ethanol was removed to prepare 125 g of ginseng flower extract powder (Examples 1 and 2).
[實施例 3] 根據加壓減壓萃取製程製備人參花萃取物以分析人參皂苷及確定抗微生物胜肽之表現 用與實施例1及2相同製備方法在24小時、水溫30°C及加壓萃取壓力為80Mpa及減壓萃取壓力為大氣壓力之條件下處理人參花(紫莖變種)以製備由加壓減壓萃取製程獲得之125g人參花萃取物(實施例3)，用以分析由該加壓減壓萃取製程獲得之萃取物之人參皂苷。[Example 3] Preparation of ginseng flower extract according to a pressurized decompression extraction process to analyze ginsenosides and determination of the performance of antimicrobial peptides The same preparation methods as in Examples 1 and 2 were carried out at 24 hours, water temperature 30 ° C and addition. The ginseng flower (purple stem variety) was treated under the conditions of pressure extraction pressure of 80 MPa and pressure extraction pressure at atmospheric pressure to prepare 125 g ginseng flower extract obtained by the pressure-pressure extraction process (Example 3) for analysis. The ginseng saponin of the extract obtained by the pressurized decompression extraction process.
[測試實施例1] 比較萃取製程間人參皂苷Re之產率及絕對量 對獲得之人參花萃取物粉末(比較實施例1及2與實施例1及2)進行HLPC分析，其條件如下： 人參皂苷Re之HPLC分析條件 - 管柱: CAPCELL PAK C18(SHISEIDO) - 偵測器(測量波長): UV吸收偵測器(203 nm) - 注射量，流速: 20 uL, 1.0 mL/min - 移動相: HPLC梯度條件(A: D.I水, B: 50% BeCN) 表1
下表2顯示人參花萃取物粉末之人參皂苷Re產率及絕對量之結果(比較實施例1及2與實施例1及2)。以100g人參花粉末為基礎計算人參皂苷Re之絕對量。 表2: 比較萃取條件間人參皂苷Re之產率及絕對量
如表2所示，相較於由一般萃取製程萃取之相同種類人參花萃取物(比較實施例1及2)，發現由加壓減壓萃取製程萃取之該人參花萃取物(實施例1及2)產率分別增加11%及10%。而且，相較於由一般萃取製程萃取之相同種類人參花萃取物(比較實施例1及2)，發現由加壓減壓萃取製程萃取之該人參花萃取物之人參皂苷Re量(實施例1及2)以100g人參花粉末為基礎分別增加0.66g及0.73g。As shown in Table 2, the ginseng flower extract extracted by the pressurized vacuum extraction process was found as compared with the same kind of ginseng flower extract extracted by the general extraction process (Comparative Examples 1 and 2) (Example 1 and 2) The yield increased by 11% and 10%, respectively. Moreover, the amount of ginsenoside Re of the ginseng flower extract extracted by the pressurized vacuum extraction process was found as compared with the same kind of ginseng flower extract extracted by the general extraction process (Comparative Examples 1 and 2) (Example 1 And 2) 0.66 g and 0.73 g, respectively, based on 100 g of ginseng flower powder.
[測試實施例2] 確定加壓減壓萃取製程之最佳條件 在測試實施例1中，發現該加壓減壓萃取製程在人參皂苷Re之產率及絕對量方面很優異。為了確認該加壓減壓萃取製程之最佳條件，人參皂苷Re之產率及絕對量(g; 以100g人參花粉末為基礎)係依序與改變之處理時間、溫度及壓力條件相比較。[Test Example 2] Determination of optimum conditions for the pressurized decompression extraction process In Test Example 1, the pressurized decompression extraction process was found to be excellent in the yield and absolute amount of ginsenoside Re. In order to confirm the optimum conditions of the pressurized decompression extraction process, the yield and absolute amount of ginsenoside Re (g; based on 100 g of ginseng flower powder) were sequentially compared with the changed treatment time, temperature and pressure conditions.
確認最佳時間條件
使用與實施例1及2製備方法相同之方法分別於1小時、6小時、12小時及24小時之30°C水溫、80MPa加壓萃取壓力及大氣壓力之減壓萃取壓力來製備根據加壓減壓萃取製程之人參花萃取物粉末。在與測試實施例1相同分析條件下進行HPLC分析。結果如表3所示。 表3: 比較人參花萃取物之人參皂苷Re隨時間改變之產率及絕對量
確認最佳溫度條件
使用與實施例1及2製備方法相同之方法以水溫分別自20°C至30°C及40°C改變於24小時、80 MPa加壓萃取壓力、大氣壓力之減壓萃取壓力條件下根據加壓減壓萃取製程製備人參花萃取物粉末。在與測試實施例1相同之分析條件下進行HPLC分析。結果如表4所示。 表4: 比較人參花萃取物之人參皂苷Re隨水溫改變之產率及絕對量
確認最佳壓力條件
使用與實施例1及2製備方法相同之方法在24小時、30°C水溫、大氣壓力之減壓萃取壓力條件下，利用自大氣壓力(與製備控制組、比較實施例1及2相同)至10MPa、20MPa、40MPa及80MPa改變之加壓萃取壓力，根據加壓減壓萃取製程製備人參花萃取物粉末。在與測試實施例1相同之分析條件下進行HPLC分析。結果如表5所示。 表5: 比較人參花萃取物之人參皂苷Re隨加壓萃取壓力改變之產率及絕對量
如表3至5所示，發現在24小時、30°C水溫、80MPa加壓萃取壓力條件下獲得人參皂苷Re之最佳產率及最高絕對量。As shown in Tables 3 to 5, it was found that the optimum yield and the highest absolute amount of ginsenoside Re were obtained under the conditions of 24 hours, 30 ° C water temperature and 80 MPa pressure extraction pressure.
[測試實施例3] 分析人參皂苷 測試實施例2中確認該加壓減壓萃取製程之最佳條件。為了分析由該加壓減壓萃取製程獲得之該萃取物之人參皂苷，在此測試中分析一般萃取製程製備之該人參花萃取物(比較實施例3)之人參皂苷含量及由該加壓減壓萃取製程製備之人參花萃取物(實施例3)。[Test Example 3] Analysis of ginsenoside The optimum conditions for the pressurized decompression extraction process were confirmed in Test Example 2. In order to analyze the ginsenoside of the extract obtained by the pressurized decompression extraction process, the ginsenoside content of the ginseng flower extract (Comparative Example 3) prepared by the general extraction process was analyzed and reduced by the pressurization in this test. Ginseng flower extract prepared by a pressure extraction process (Example 3).
在與測試實施例1相同分析條件下對比較實施例3及實施例3之人參花萃取物進行HPLC分析。結果如圖1所示。下表6顯示由以上結果獲得之在該人參花萃取物中人參皂苷各成分之絕對量(g; 以100g人參花粉末為基礎)。 [表6] 比較分析由一般萃取製程及該加壓減壓萃取製程製備之該人參花萃取物之人參皂苷各成分之絕對量(單位:g)
所表6所示，發現由加壓減壓萃取製程獲得之該人參花萃取物(實施例 3)包含較由一般萃取製程獲得之該人參花萃取物(比較實施例3)多約30至112%之人參皂苷成分。也發現實施例3之人參花萃取物所含之人參皂苷總重顯著高於比較實施例3。As shown in Table 6, it was found that the ginseng flower extract obtained by the pressurized vacuum extraction process (Example 3) contained about 30 to 112 more than the ginseng flower extract obtained by the general extraction process (Comparative Example 3). % of ginsenosides. The total weight of ginsenoside contained in the ginseng flower extract of Example 3 was also found to be significantly higher than that of Comparative Example 3.
[測試實施例4] 確認抗微生物胜肽基因之表現 為了確認在測試實施例2最佳條件下製備之該人參花萃取物對抗微生物胜肽基因表現之效果，進行以下測試。於該測試中使用之該抗微生物胜肽基因為hBD2(人類β防禦素2)、hBD3(人類β防禦素3)及牛皮癬素(psoriasin)。[Test Example 4] Confirmation of the performance of the antimicrobial peptide gene In order to confirm the effect of the ginseng flower extract prepared under the optimum conditions of Test Example 2 against the performance of the microbial peptide gene, the following test was conducted. The antimicrobial peptide genes used in this test were hBD2 (human beta defensin 2), hBD3 (human beta defensin 3), and psoriasin.
在KGM-gold(Lonza, # 00192151)培養基中讓角質細胞生長，然後將25,000個角質細胞加入至6-井盤培養24小時。以95°C熱水萃取法獲得之人參花萃取物(比較實施例4)、以一般萃取法在一大氣壓下獲得之人參花萃取物 (比較實施例3)及以80MPa加壓減壓萃取獲得之該人參花萃取物(實施例3)在培養基中稀釋至40ppm然後施用至該角質細胞。控制組(無處理組)未經處理。在反應24小時後，將1mL之trizol加入至該等細胞以萃取RNA，於室溫進行均質化5分鐘。將該混合物移至一1.5 mL管，將0.1 mL氯仿加入其中。用手振動該混合物15秒，於室溫反應15分鐘然後於4°C以13,000 rpm離心15分鐘。將0.4 mL透明上清液移置一新管中，將0.5 mL異丙醇加入其中，用手振動該混合物15秒。於室溫反應8分鐘後，於4°C以13,000 rpm離心該混合物8分鐘。丟棄該上清液，將0.5 mL之75%乙醇加入至所獲得之沉澱物，接著於4°C以10,000 rpm離心5分鐘。丟棄該上清液，將22 μL去離子水加入至所獲得之沉澱物以溶解純RNA。使用分光光度計測量260nm吸光度以決定該RNA量。然後，將4μg之RNA、3.5 μL之10× RT緩衝液(Invitrogen, #18080-051)、2 μL之50 μM寡核苷酸dT、3.5μL之0.1M DTT、7μL之25mM MgCl2 、3μL之10mM dNTP、1μL之RNAaseOutTM 及1μL之Superscript III混合以合成cDNA。然後，將去離子水加入以達到35 μL。使用PCR儀器於50°C進行RT反應60分鐘。將165 μL去離子水加入至35 μL該所得之cDNA混合物以達到總計200 μL。為了進行定量即時聚合酶鏈鎖反應(real-time qPCR)，將5 μL之 cDNA混合物、4 μL去離子水、10 μL之Taqman 2X Universal PCR Master Mix (ABI, #4304437)及1 μL之Taqman primer(控制組基因: RPLP0，測試組基因: hBD2、hBD3、牛皮癬素(psoriasin))混合以達到總計20 μL。然後，將該溶液加入至一96-井PCR管之各井，然後將其加載至即時聚合酶鏈鎖反應儀器中(ABI, model: 7500fast)。該反應設定如下：1. 保持階段(Holding phase): 95°C (10分鐘); 2. 循環階段(Cycling phase): 95°C (15秒)及60°C (60秒)，反復40次循環。於此測試中使用之Taqman primer係一商購引子，其係由指示螢光分子(reporter fluorescent molecule)FAM及淬滅體(quencher)NFQ-MGB組成。hBD2、hBD3及牛皮癬素(psoriasin)基因表現量之比較結果係如圖2所示。Keratinocytes were grown in KGM-gold (Lonza, # 00192151) medium, and then 25,000 keratinocytes were added to the 6-well plate for 24 hours. The ginseng flower extract obtained by the hot water extraction method at 95 ° C (Comparative Example 4), the ginseng flower extract obtained by the general extraction method under atmospheric pressure (Comparative Example 3) and the pressure-extraction extraction at 80 MPa were obtained. The ginseng flower extract (Example 3) was diluted to 40 ppm in the medium and then applied to the keratinocytes. The control group (no processing group) was not processed. After 24 hours of reaction, 1 mL of trizol was added to the cells to extract RNA, which was homogenized for 5 minutes at room temperature. The mixture was transferred to a 1.5 mL tube and 0.1 mL of chloroform was added thereto. The mixture was shaken by hand for 15 seconds, reacted at room temperature for 15 minutes and then centrifuged at 13,000 rpm for 15 minutes at 4 °C. 0.4 mL of the clear supernatant was placed in a new tube, 0.5 mL of isopropanol was added thereto, and the mixture was shaken by hand for 15 seconds. After reacting for 8 minutes at room temperature, the mixture was centrifuged at 13,000 rpm for 8 minutes at 4 °C. The supernatant was discarded, and 0.5 mL of 75% ethanol was added to the obtained precipitate, followed by centrifugation at 10,000 rpm for 5 minutes at 4 °C. The supernatant was discarded, and 22 μL of deionized water was added to the obtained precipitate to dissolve the pure RNA. The absorbance at 260 nm was measured using a spectrophotometer to determine the amount of RNA. Then, 4 μg of RNA, 3.5 μL of 10× RT buffer (Invitrogen, #18080-051), 2 μL of 50 μM oligonucleotide dT, 3.5 μL of 0.1 M DTT, 7 μL of 25 mM MgCl 2 , 3 μL 10mM dNTP, 1μL of RNAaseOut TM [mu] L of Superscript III and mixed to synthesize cDNA. Then, deionized water was added to reach 35 μL. The RT reaction was carried out at 50 ° C for 60 minutes using a PCR instrument. 165 μL of deionized water was added to 35 μL of the resulting cDNA mixture to achieve a total of 200 μL. For quantitative real-time qPCR, 5 μL of cDNA mixture, 4 μL of deionized water, 10 μL of Taqman 2X Universal PCR Master Mix (ABI, #4304437) and 1 μL of Taqman primer (Control group gene: RPLP0, test group genes: hBD2, hBD3, psoriasin) were mixed to achieve a total of 20 μL. This solution was then added to each well of a 96-well PCR tube and loaded into an instant polymerase chain reaction instrument (ABI, model: 7500fast). The reaction was set as follows: 1. Holding phase: 95 ° C (10 minutes); 2. Cycling phase: 95 ° C (15 seconds) and 60 ° C (60 seconds), repeated 40 times cycle. The Taqman primer used in this test is a commercially available primer consisting of a reporter fluorescent molecule FAM and a quencher NFQ-MGB. The results of the comparison of hBD2, hBD3 and psoriasin gene expression are shown in Fig. 2.
如圖2所示，發現相較於未處理組(控制組)、熱水萃取(比較實施例4)及由一般萃取方式在一大氣壓下獲得之萃取物(比較實施例3)，由加壓減壓萃取製程以80MPa加壓萃取壓力獲得之人參花萃取物(實施例3)展現出抗微生物胜肽基因hBD2、hBD3及牛皮癬素(psoriasin)之增加表現量。因此，發現由加壓減壓萃取在最佳條件下製備之含有特定量之人參皂苷之人參花萃取物能藉由增加抗微生物胜肽表現來提升皮膚屏障功能。As shown in Fig. 2, it was found that the extract was obtained by pressurization (Comparative Example 3) compared to the untreated group (control group), hot water extraction (Comparative Example 4), and the general extraction method at atmospheric pressure (Comparative Example 3). The ginseng flower extract obtained by the pressure-reduction extraction process at a pressure of 80 MPa under pressure extraction (Example 3) exhibited an increased expression amount of the antimicrobial peptide genes hBD2, hBD3, and psoriasin. Therefore, it has been found that a ginseng flower extract containing a specific amount of ginseng saponin prepared under optimal conditions by pressurized decompression extraction can enhance the skin barrier function by increasing the performance of the antimicrobial peptide.
以下根據本發明一態樣之組成物之製劑實施例將被描述。然而，其可被配製成各種其他形式。此等實施例僅用於說明目的而非有意限制本發明之範圍。The following formulation examples of the composition according to one aspect of the present invention will be described. However, it can be formulated in various other forms. The examples are for illustrative purposes only and are not intended to limit the scope of the invention.
[製劑實施例1] 保濕劑 根據習知方法以下表7所示組成物製備保濕劑。 表7
[製劑實施例2] 滋潤乳液(lotion) 根據習知方法以下表8所示組成物製備滋潤乳液。 表8
[製劑實施例3] 滋養霜 根據習知方法以下表9所示組成物製備滋養霜。 表9
[製劑實施例4] 髮膜(Hair pack) 根據習知方法以下表10所示組成物製備髮膜。 表10
[製劑實施例5] 軟膏性 根據習知方法以下表11所示之組成物製備一軟膏劑。 表11
製劑實施例6: 局部施用之藥品(貼布) 根據習知方法以下表12所示組成物製備局部施用之藥品(貼布)。 表12
[製劑實施例7] 製備散劑(powders) 實施例3之人參花萃取物 2 g 乳糖 1 g 以上成分被混合及填入一氣密囊袋以製備散劑。[Formulation Example 7] Preparation of powders Ginseng flower extract of Example 3 2 g Lactose 1 g The above ingredients were mixed and filled into an airtight pouch to prepare a powder.
[製劑實施例8] 製備錠劑 實施例3之人參花萃取物 100 mg 玉米澱粉 100 mg 乳糖 100 mg 硬脂酸鎂 2 mg 以上成分被混合及根據習知製備錠劑之方法製錠以製備成錠劑。[Formulation Example 8] Preparation of a tablet ginseng flower extract of Example 3 100 mg Corn starch 100 mg Lactose 100 mg Magnesium stearate 2 mg The above ingredients were mixed and prepared in accordance with a conventional method for preparing a tablet. Lozenges.
[製劑實施例9] 製備膠囊 實施例3之人參花萃取物 100 mg 玉米澱粉 100 mg 乳糖 100 mg 硬脂酸鎂 2 mg 以上成分被混合及根據習知製備膠囊之方法填入明膠膠囊以製備成膠囊。[Formulation Example 9] Preparation of capsules Ginseng flower extract of Example 3 100 mg Corn starch 100 mg Lactose 100 mg Magnesium stearate 2 mg The above ingredients were mixed and filled into gelatin capsules according to a conventional method of preparing capsules to prepare into capsule.
[製劑實施例10] 製備丸劑 實施例3之人參花萃取物 1 g 乳糖 1.5 g 甘油 1 g 木糖醇 0.5 g 以上成分被混合及根據習知方法製備成一重4g之丸劑。[Formulation Example 10] Preparation of Pills Ginseng Flower Extract of Example 3 1 g Lactose 1.5 g Glycerol 1 g Xylitol 0.5 g The above components were mixed and prepared into a single 4 g pellet according to a conventional method.
[製劑實施例11] 製備顆粒劑 實施例3之人參花萃取物 150 g 大豆萃取物 50 mg 葡萄糖 200 mg 澱粉 600 mg 以上成分被混合、加入100mg之30%乙醇及在60°C乾燥以形成一顆粒劑，而所得之顆粒劑被填入一囊袋(pouch)。[Formulation Example 11] Preparation of granules Ginseng flower extract of Example 3 150 g Soybean extract 50 mg Glucose 200 mg Starch 600 mg The above ingredients were mixed, added with 100 mg of 30% ethanol and dried at 60 ° C to form a Granules, and the resulting granules are filled into a pouch.
[製劑實施例12] 飲料 50 mg實施例3之人參花萃取物、10 g葡萄糖、0.6 g檸檬酸及25 g液體寡糖被混合及加入300 ml純水。200 ml所得混合物被填入各瓶中。在填入瓶中之後，將混合物在130℃下滅菌4至5秒以製備飲料。[Formulation Example 12] Beverage 50 mg of the ginseng flower extract of Example 3, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide were mixed and 300 ml of pure water was added. 200 ml of the resulting mixture was filled into each bottle. After filling in the bottle, the mixture was sterilized at 130 ° C for 4 to 5 seconds to prepare a beverage.
[製劑實施例13] 焦糖製劑 一焦糖製劑可藉由混合50 mg實施例3之人參花萃取物、1.8 g玉米糖漿、0.5 g脫脂牛奶、0.5 g大豆卵磷脂、0.6 g奶油、0.4 g植物氫化油、1.4 g糖、0.58 g人造黃油及20 mg食鹽來形成。[Formulation Example 13] Caramel Formulation One caramel preparation can be prepared by mixing 50 mg of the ginseng flower extract of Example 3, 1.8 g of corn syrup, 0.5 g of skim milk, 0.5 g of soybean lecithin, 0.6 g of cream, 0.4 g. It is formed by vegetable hydrogenated oil, 1.4 g of sugar, 0.58 g of margarine and 20 mg of salt.
無no
圖1係一圖表，其比較以一般萃取製程萃取之人參花萃取物之人參皂苷 (大氣壓條件)(比較實施例3)與該等以本發明一具體實施例之萃取製程萃取之人參花萃取物(加壓萃取壓力80 MPa)(實施例3)。 圖2係一圖表，其顯示在未處理組(控制組)、以熱水萃取法萃取之人參花萃取物(比較實施例4)、以一般萃取製程(大氣壓條件)萃取之人參花萃取物(比較實施例3)及以本發明一具體實施例之萃取製程(加壓萃取壓力80 MPa)萃取之人參花萃取物(實施例3)之間hBD2(圖2A)、hBD3(圖2B)及牛皮癬素(psoriasin)(圖2C)基因表現之比較結果。Figure 1 is a graph comparing ginsenosides (atmospheric conditions) of a ginseng flower extract extracted by a general extraction process (Comparative Example 3) with the ginseng flower extracts extracted by the extraction process of a specific embodiment of the present invention. (Pressure extraction pressure 80 MPa) (Example 3). Figure 2 is a graph showing ginseng flower extract extracted in an untreated group (control group), ginseng flower extract extracted by hot water extraction (Comparative Example 4), and a general extraction process (atmospheric pressure). Comparative Example 3) and hBD2 (Fig. 2A), hBD3 (Fig. 2B) and psoriasis extracted between the ginseng flower extract (Example 3) extracted by the extraction process (pressure extraction pressure 80 MPa) of one embodiment of the present invention The results of the comparison of gene expression of psoriasin (Fig. 2C).
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| JP5628797B2 (en) * | 2008-06-13 | 2014-11-19 | アモーレパシフィック コーポレイションＡｍｏｒｅｐａｃｉｆｉｃ Ｃｏｒｐｏｒａｔｉｏｎ | Skin external preparation composition containing ginseng flower or ginseng seed extract |
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