WO2015024396A1 - 保湿祛皱及舒敏功效的植物组合物及其制备方法 - Google Patents
保湿祛皱及舒敏功效的植物组合物及其制备方法 Download PDFInfo
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- WO2015024396A1 WO2015024396A1 PCT/CN2014/077149 CN2014077149W WO2015024396A1 WO 2015024396 A1 WO2015024396 A1 WO 2015024396A1 CN 2014077149 W CN2014077149 W CN 2014077149W WO 2015024396 A1 WO2015024396 A1 WO 2015024396A1
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- Prior art keywords
- oat
- water
- composition
- chinese medicine
- traditional chinese
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Classifications
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
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- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/86—Violaceae (Violet family)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8968—Ophiopogon (Lilyturf)
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- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
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- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
Definitions
- the invention relates to the field of cosmetic technology development, and particularly relates to a plant composition for moisturizing wrinkle and sedative effect and a preparation method thereof.
- the skin has sufficient water content and is the basis for physiological functions such as skin barrier, absorption, and metabolism. Adequate hydration is beneficial to the enzyme reaction and promotes the ripening of the stratum corneum, maintaining the elasticity of the stratum corneum; facilitating the expansion of the stratum corneum cells themselves, the density of the structure is reduced, and the permeability of the material is increased; Regular reflections create a bright luster, while a dry, scaly stratum corneum reflects light in a non-specular reflection, making the skin gray. Therefore, most women today focus their skin care on skin moisturizing, and moisturizing has become a top priority in skin care products.
- the ability of the skin to retain water is primarily related to the stratum corneum, which acts as a barrier to water loss.
- stratum corneum there are water-soluble moisturizing substances such as amino acid or salt, sugar (NMF, also known as natural moisturizing factor), and oily components such as cell lipids and sebum (the epidermal sebum present in the epidermis of the skin), among which natural Moisture factor accounts for 30%, and oily ingredients account for 11%.
- NMF also known as natural moisturizing factor
- oily components such as cell lipids and sebum (the epidermal sebum present in the epidermis of the skin), among which natural Moisture factor accounts for 30%, and oily ingredients account for 11%.
- These oily ingredients combine with natural moisturizing factors, or surround natural moisturizing factors to prevent them from flowing out, and play an appropriate role in controlling water volatilization.
- Aquaporin 3 is the most abundant water channel in the skin. RT-PCR method was used to find that AQP3 is most abundant in human epidermal keratinocytes and stratum corneum. AQP3 is of great significance for the water retention of keratinocytes.
- the phenotypic observation of the AQP3 knockout hairless mouse showed that the development of the mouse was basically normal, but the water content of the epidermal keratinocytes of AQP3 knockout mice was significantly reduced, and the amount of water loss when exposed to dryness was also significantly higher than normal. Mouse.
- Health, natural is the slogan of beauty business. China has a long history of manufacturing and using natural botanical cosmetics, rich practical experience and technological advantages, and China's vast land and natural resources, in its unique geographical environment and climate conditions, grow a number of unique natural plants. These plants have long been used for beauty because of their different characteristics, which moisturize the skin and prevent various harmful factors from affecting the skin.
- This patent is based on this, from the development of natural plants to moisturizing effect, and has anti-aging and anti-sensitive cosmetic additives, extracting from a variety of plants, through film formation, water retention, maintenance of normal aquaporin Physiological functions and other ways, synergistically, to achieve a full range of moisturizing anti-aging anti-sensitive effects. It is an object of the present invention to provide a botanical composition for moisturizing wrinkle and soothing efficacy and a process for the preparation thereof.
- the traditional Chinese medicine composition provided by the invention has an active ingredient which is an aqueous extract of the following raw material mixture - Dendrobium, Pansy, Ophiopogon japonicus, Tremella and oat raw materials;
- the oat raw material is oatmeal, oat flour or oat bran.
- the oat granules are oat granules with or without bran;
- the oat flour is whole seed oat flour or oat flour; the whole seed oat flour is crushed after the whole seed oats are crushed
- the mass ratio of the raw materials of Dendrobium, Pansy, Ophiopogon japonicus, Tremella and oatmeal is the mass ratio of the raw materials of Dendrobium, Pansy, Ophiopogon japonicus, Tremella and oatmeal.
- the mass ratio of the water to the sarcophagus in the water extraction step is 160-220: 5-15, specifically 160: 12, 220: 5, 200: 12, 160-200: 12, 200-220: 5-12, 160-220: 5-12.
- the aqueous extract is prepared according to the method comprising the following steps:
- the stone scorpion, pansy, Ophiopogon japonicus, tremella, oat raw material and water are mixed and extracted according to the ratio; in the extraction step, the temperature is 60-100 ° C, specifically 65 ° C, 80 ° C, 100 ° C, 65 ° C - 100 ° C, 80 ° C - 100 ° C or 65-80 ° C; time is 1-4 hours, specifically 2 hours.
- the method for preparing the composition provided by the present invention comprises the steps of: mixing the sarcophagus, pansy, Ophiopogon japonicus, tremella, oat raw material and water according to the formula according to claim 2 or 3, and then extracting Water extract.
- the temperature is 60-100 ° C, specifically 80 ° C; the time is 1-4 hours, and the specific time is 1, 2, 4, 1-2 hours or 2-4 hours.
- the method further comprises the step of - after the extraction step, the resulting aqueous extract is allowed to stand, filtered, and the filtrate is collected.
- the temperature is room temperature
- the time is 1-4 hours, specifically 1, 2, 4 hours
- the mesh number of the filtering holes is 100-200 mesh.
- the invention adopts the combination of Dendrobium, Pansy, Ophiopogon, oat raw material and Tremella fuciformis, and then extracts it for external use, and maintains the moisturizing effect and delays skin aging and anti-sensitivity by holding water and maintaining the physiological function of normal aquaporin. And anti-cracking effect.
- Figure 1 is a graph showing the percentage of water content on the surface of the skin as a function of time.
- Figure 2 is a graph showing the amount of water loss on the surface of the skin as a function of time.
- Figure 3 shows the effect of the composition on DPPH removal.
- Figure 4 shows the inhibitory effect of the composition on hyaluronidase.
- the present invention will be further described below in conjunction with specific embodiments, but the invention is not limited to the following examples.
- the method is a conventional method unless otherwise specified.
- the raw materials can be obtained from an open commercial route unless otherwise specified.
- Example 1 The raw materials can be obtained from an open commercial route unless otherwise specified.
- the traditional Chinese medicine composition was prepared as follows:
- Example 2 Two layers of filter paper were placed in a Buchner funnel, and about 0.5 cm of diatomaceous earth was sandwiched between the filter papers, and the filtrate obtained by coarse filtration was vacuum-filtered, and the filtrate was collected to obtain a traditional Chinese medicine composition A provided by the present invention.
- Example 2 Two layers of filter paper were placed in a Buchner funnel, and about 0.5 cm of diatomaceous earth was sandwiched between the filter papers, and the filtrate obtained by coarse filtration was vacuum-filtered, and the filtrate was collected to obtain a traditional Chinese medicine composition A provided by the present invention.
- Example 2 Two layers of filter paper were placed in a Buchner funnel, and about 0.5 cm of diatomaceous earth was sandwiched between the filter papers, and the filtrate obtained by coarse filtration was vacuum-filtered, and the filtrate was collected to obtain a traditional Chinese medicine composition A provided by the present invention.
- Example 2 Two layers of filter paper were placed in a Buchner funnel, and about 0.5 cm of diatomaceous earth was sandwiched between the filter papers
- the traditional Chinese medicine composition was prepared according to the following steps -
- the traditional Chinese medicine composition is prepared according to the following steps -
- Example 4 Example 1, Example 2, Example 3 Evaluation of moisturizing effect of traditional Chinese medicine compositions A, B, and C
- the test sample is a cream, which is prepared according to the following method - the traditional Chinese medicine composition of any of Examples 1-3 is added to the phase B of the blank cream base to obtain a cream, wherein the quality of the traditional Chinese medicine composition in the cream is 100 The content of each part is 5%.
- the traditional Chinese medicine composition A obtained in the first embodiment corresponds to the cream 1
- the traditional Chinese medicine composition B obtained in the second embodiment corresponds to the cream 2
- the traditional Chinese medicine composition C obtained in the third embodiment corresponds to the cream 3.
- a blank cream base was also used as a blank control.
- the preparation method of the blank cream base used is as follows - according to the formulation obtained in Table 1, the raw materials of phase A are heated, uniformly mixed, and heated to 82; the raw materials of phase B are heated, uniformly mixed, and heated to 82 ° C; Homogenize at 2000r/min, phase A slowly into phase B, homogenize for 5min; after homogenization is completed, stir and cool; 40° when phase C is added, stir evenly, that is, a blank cream matrix is obtained.
- the test site should be at room temperature (25 ⁇ 1) V and relative humidity (40 ⁇ 5)%. Subjects should maintain the body in a stable state before testing. After wiping the subject's arms with about 35 ounces of water, the test was started after sitting for 30 minutes in the test environment.
- the affected area (4 C mx4 C m) was marked on the inside of the subject's arm 5 cm from the base of the palm, and multiple areas (interval lcm) were simultaneously marked on the same arm.
- the test samples were randomly distributed. The blank value of each test area was measured first, and then the test sample was applied in a single application at a dose of (2.0 ⁇ 0.1) mg/cm 2 . After coating, the multi-purpose skin moisture tester and the skin water disperse meter were respectively measured for 0.5 h. Skin hydration status values and water loss loss values of the test area and the blank control area at lh, 2 h, and 4 h.
- test averaged the data measured 5 times in the same sample area and used the SPSS statistical software for t-test analysis.
- the change of moisture content is reflected in the test period.
- Hydration rate (sample group moisture content - initial moisture content) / initial moisture content
- Transepidermal water loss reflects the change of water dispersion in the experimental area over time during the test period. It can characterize the water retention function of the test sample. The smaller the value, the less the water dispersion loss, the water retention capacity. The stronger; on the contrary, the weaker the ability to lock water.
- the traditional Chinese medicine composition provided by the invention can increase the moisture content of the skin, reduce the water dispersion loss, has a good water-locking ability, and has a significant moisturizing effect on the skin.
- Example 5 Example 1, Example 2, Example 3 obtained Chinese medicine composition A, B, C for AQP3 mRNA The influence of the expression volume
- Aquaporins are a group of membrane transport proteins involved in the transmembrane transport of water, glycerol and some small molecules.
- AQP3 is expressed in a variety of tissues and organs including the skin. Normal epidermal KC expresses AQP3 only in the basal cell layer.
- AQP3 provides a channel for epithelial cell membranes, maintains intracellular osmotic pressure and cell volume, and plays a role in water transport.
- the glycerol transport function of AQP3 plays a key role in skin hydration, elastic retention and repair function after injury.
- composition of this study can play a role in repairing the skin barrier and increasing the water content of the stratum corneum through a moisturizing route, and may also exert these effects by regulating the level of AQP3.
- traditional Chinese medicine composition A, B, C solution to C3H mice The expression of AQP3 mRNA in mouse skin was detected by real-time quantitative PCR and compared with normal saline.
- Example 1 sample group A A total of 20 experimental mice were divided into Example 1 sample group A, Example 2 sample group B, and Example 3 sample group C, negative control group, 5 rats in each group.
- Sample group The obtained Chinese medicine composition of any of Examples 1-3 and deionized water were mixed at a mass ratio of 5:95 to obtain an aqueous solution of the traditional Chinese medicine composition, and the aqueous solution of the traditional Chinese medicine composition was separately applied to the experimental mice. , apply once a day;
- Control group Apply saline once a day, that is, to maintain normal physiological state of the skin.
- RNA in keratinocytes was extracted with Trizol and reverse transcribed into cDNA by real-time quantitative PCR SYBRGREEN method (QIAGEN 204143).
- the primer sequence and conditions of PCR are as follows: aqp3 sense : 5'-TTGGTGGCTGGCCAAGTGTC-3'; aqp3
- Antisense 5' -GTCTGTGCC AGTGC ATAGAT-3 ' ;
- B- actin sense 5' -GTCTGTGCC AGTGC ATAGAT-3 ' ;
- the traditional Chinese medicine composition has the effect of up-regulating the expression of AQP3 mRNA, which enhances skin hydration by stimulating the transport of stratum corneum water or glycerol, and moisture-binding molecules (such as hyaluronic acid) in the skin. Increased production, reduced skin dryness and shedding.
- Example 6 Example 1, Example 2, Example 3 Anti-aging function evaluation of the traditional Chinese medicine compositions A, B, and C
- DPPH (1, l-disphenyl-2-picryl-hydrazyl), chemically named 1, 1 diphenyl-2-trinitrophenylhydrazine, is a stable organic free radical. Quantitative analysis by spectrophotometry Example 1, Example 2, Example 3 The traditional Chinese medicine composition A, B, C were used to remove free radicals.
- the obtained traditional Chinese medicine composition of any of Examples 1-3 and deionized water are mixed at a mass ratio of 5:95 to obtain an aqueous solution of the traditional Chinese medicine composition;
- Example 1 the traditional Chinese medicine composition A obtained in Example 1 corresponds to sample 1;
- the traditional Chinese medicine composition B obtained in Example 2 corresponds to sample 2 ;
- the obtained Chinese medicine composition C of Example 3 corresponds to sample 3; the traditional Chinese medicine composition in the test group was replaced with a vitamin C derivative, and the obtained aqueous solution was used as a positive control; the measurement result is shown in Fig. 3.
- the experimental results show that when the mass percentage of the traditional Chinese medicine composition obtained in each example is 5.0% in its aqueous solution, the uniformity has the ability to scavenge DPPH, and has an effective concentration for reducing hydroxyl radicals, alkyl radicals or oxidative radicals. It interrupts the action of lipid peroxidation chain reaction and thus has a good effect of delaying skin photoaging.
- the effect of the traditional Chinese medicine composition on the activity of MMP-1 was examined by the fluorogenic method.
- the experimental method refers to "two kinds of Explain the experimental methods described in the book Establishment and Application of the In-situ Screening Method for Deteriorating Effect Additives (To Jixiang, 2007).
- the obtained traditional Chinese medicine composition of any of Examples 1-3 and deionized water are mixed at a mass ratio of 5:95 to obtain an aqueous solution of the traditional Chinese medicine composition;
- Example 1 Composition A 81.63
- Example 3 Composition C 60.98 The results show that each of the composition compositions has an inhibitory effect on MMP-1 induced by infrared radiation, which can reduce excessive degradation of collagen and elastin caused by light, and maintain the skin. Resilience, reduce the role of skin photoaging.
- Example 7, Example 1, Example 2, Example 3 The anti-allergic effect of the obtained traditional Chinese medicine composition A, B, C
- Hyaluronic acid is a component of the tissue matrix that restricts the diffusion of water and other extracellular substances. After hydrolysis by hyaluronidase, the cells become non-tacky, and the new medium of cell degranulation and synthesis oozes out, exerting biological effects. Lead to the occurrence of immediate allergic reactions. Therefore, the degree of inhibition of hyaluronidase by the commonly used samples reflects the alleviation and improvement of the type I hypersensitivity reaction.
- Example 1 The degree of inhibition of hyaluronidase by the traditional Chinese medicine compositions obtained in Example 1, Example 2, and Example 3, B and C was measured according to the following method - the Chinese medicine composition A and the deionized water obtained in Example 1 were 5:95. The mass ratio is mixed to obtain a sample solution
- the traditional Chinese medicine composition B obtained in Example 2 and deionized water were mixed at a mass ratio of 5:95 to obtain a sample solution.
- Example 3 The traditional Chinese medicine composition C obtained in Example 3 and deionized water were mixed at a mass ratio of 5:95 to obtain a sample solution.
- the test was set up three times.
- the method of processing each repetition is as follows:
- the B tube is added as a control blank solution, corresponding to column B in Table 4.
- the D tube is added as a sample blank solution, corresponding to column D in Table 4.
- the E tube is added as a positive control solution, corresponding to column E in Table 4.
- the C1-C3 tubes are all added with reagents in the order of column C in Table 4;
- the sample solution added by the C1 tube is the sample solution C1;
- the sample solution added by the C2 tube is the sample solution C2 ;
- the sample solution added by the C3 tube is the sample solution C3;
- Hyaluronidase inhibition rate of positive control [ (A-B ) - (E-D)] / (A-B) x lOO%
- A is the ABS value of the A pipe
- B is the ABS value of the B tube
- C is the ABS value of the C tube
- D is the ABS value of the D tube
- E is the ABS value of the E tube; wherein the ABS value of the C tube is determined as follows -
- the hyaluronidase inhibition rate of the composition A obtained in Example 1 was 75.32%;
- the hyaluronidase inhibition rate of the composition B obtained in Example 2 was 59.89%;
- the hyaluronic acid inhibition rate of the composition C obtained in Example 3 was 67.26%.
- the hyaluronidase inhibition rate of the E tube i.e., the positive control, was 92.12%.
- Example 8 Example 1, Example 2, Example 3 obtained Chinese medicine composition A, B, C anti-cracking effect
- the group A is applied to the composition cream of the embodiment 1;
- composition cream the preparation method of the blank cream base used is as follows:
- the raw materials of phase A are heated, mixed uniformly, and heated to 82 ° C; the raw materials of phase B are heated, mixed uniformly, heated to 82 ° C; in phase B at 2000 r / min Quality, slowly phase A into phase B, homogenize for 5min; after homogenization is completed, stir and cool down; add 40 phase at 40 ,, stir evenly, that is, get blank cream base.
- Example 1 A 21 7 2 23 7 0 22 7 1
- Example 2 Composition B 15 12 3 25 5 0 18 10 2
- Example 3 Composition C 17 11 2 25 5 0 20 10 0 Blank control 11 15 4 20 9 1 17 7 6
- the results of the survey confirmed that the traditional Chinese medicine composition has the following characteristics:
- the active ingredients of Chinese herbal medicine contained in the product can enhance the body's self-defense ability and stimulate the growth of granulation tissue.
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Abstract
一种保湿祛皱及舒敏的中药组合物及其制备方法。该中药组合物的活性成分为石斛、三色堇、麦冬、银耳和燕麦原料的水提液。其中所述燕麦原料为燕麦粒、燕麦粉或燕麦麸皮。所述组合物中石斛、三色堇、麦冬、银耳和燕麦的质量比为5-15:2-10:2-10:1-5:1-5,优选为12:6:3:2:2。石斛、三色堇、麦冬、燕麦原料和银耳组合提取后外用,通过持水、维持正常水通道蛋白生理功能等途径,达到保湿作用同时兼具延缓皮肤衰老、抗敏感及防皲裂的功效。
Description
说 明 书 保湿祛皱及舒敏功效的植物組合物及其制备方法 技术领域
本发明涉及化妆品技术开发领域, 具体涉及一种保湿祛皱及舒敏功效的植物组合 物及其制备方法。
背景技术
现代人皮肤瘙痒、 起皱、 敏感等皮肤干燥现象严重。 皮肤水含量充足, 是皮肤屏 障、 吸收、 代谢等生理功能的基础。 充分的水合作用有利于酶反应并可促进角质层成 熟, 保持角质层的弹性;有利于角质层细胞自身发生膨胀, 结构的致密程度降低, 物质 的渗透性增加; 含水较多的角质层有规则的反射可形成明亮的光泽, 而干燥、 有鳞屑 的角质层以非镜面反射的形式反射光线, 使皮肤灰暗。 因此, 如今绝大部分的女性将 护肤重点集中到了皮肤保湿上, 保湿功效也成为护肤品中的重中之重。
皮肤保持水的能力主要和角质层有关, 角质层对水损失起了屏障作用。 角质层存 在氨基酸或盐、糖等水溶性的保湿物质 (natural moisturizing factor, 简称 NMF, 也称自 然保湿因子)和细胞脂质、皮脂等油性成分(存在于皮肤表皮的称表皮皮脂),其中天然 保湿因子占 30%, 油性成分占 11%, 这些油性成分与天然保湿因子相结合, 或包围天 然保湿因子防止其流出, 对水分挥发起着适当的控制作用。 此外, 皮肤中还存在着若 干水通道影响皮肤水分运输。 水通道蛋白 3 (AQP3 )是皮肤中表达最多的水通道, 使 用 RT-PCR方法发现 AQP3在人表皮角质细胞和角质层中含量都最为丰富。 AQP3对 角质细胞的水保持具有十分重要的意义。对 AQP3基因敲除的无毛小鼠表型观察发现, 该小鼠发育基本正常, 但 AQP3基因敲除小鼠的表皮角质细胞含水量显著降低, 暴露 于干燥情况下水损失数量也明显高于正常小鼠。
"健康、 天然"是美容事业的口号。 中国有着制造和使用天然植物化妆品的悠久 历史, 丰富的实践经验和技术优势, 而且中国地大物博, 在其独特的地理环境及气候 条件下, 生长着许多各具特色的天然植物。 这些植物因其不同的特性,长期以来用于美 容,具有滋润肌肤,防止外界各种有害因素对肌肤伤害的作用。
本专利正是基于此, 从天然植物中开发以保湿功效为主, 并且具有抗衰老与抗敏 感作用的化妆品添加剂, 对多种植物提取复配, 通过成膜、 持水、 维持正常水通道蛋 白生理功能等途径, 协同增效, 到达全方位的保湿抗衰老抗敏感效果。 本发明的目的是提供一种保湿祛皱及舒敏功效的植物组合物及其制备方法。
本发明提供的中药组合物, 其活性成分为如下原料混合物的水提液- 石斛、 三色堇、 麦冬、 银耳和燕麦原料;
说 明 书 其中, 所述燕麦原料为燕麦粒、 燕麦粉或燕麦麸皮。
上述组合物中, 燕麦粒为带麸皮或不带麸皮的燕麦粒;
所述燕麦粉为全籽燕麦粉或燕麦精粉; 所述全籽燕麦粉为将全籽燕麦粉碎后过
100目筛而得。
所述石斛、 三色堇、 麦冬、 银耳和燕麦原料的质量比为
5-15: 2-10: 2-10: 1-5: 1-5, 具体为 12: 6: 3: 2: 2、 5: 8: 2: 1: 1、 12: 3: 6: 5: 4、 5-12: 3-8: 1-5: 1-4、 12: 3-6: 2-5: 2-4;
所述水提液中, 水提步骤所用水与石斛的质量比为 160-220: 5-15, 具体为 160: 12、 220: 5、 200: 12、 160-200: 12、 200-220: 5-12、 160-220: 5-12。
所述水提液为按照包括如下歩骤的方法制备而得:
将所述石斛、 三色堇、 麦冬、 银耳、 燕麦原料和水按配比混匀后提取而得; 所述提取歩骤中, 温度为 60-100°C, 具体为 65°C、 80°C、 100°C、 65°C-100°C、 80°C-100°C或 65-80°C ; 时间为 1-4小时, 具体为 2小时。
本发明提供的制备所述组合物的方法, 包括如下步骤- 将所述石斛、 三色堇、 麦冬、 银耳、 燕麦原料和水按照权利要求 2或 3所述配比 混匀后于提取得到水提液。
上述方法所述提取歩骤中, 温度为 60-100°C, 具体为 80°C ; 时间为 1-4小时, 具 体为 1、 2、 4、 1-2小时或 2-4小时。
所述方法还包括如下步骤- 在所述提取歩骤之后, 将所得水提液静置后过滤, 收集滤液。
其中, 所述静置步骤中, 温度为室温, 时间为 1-4小时, 具体为 1、 2、 4小时; 过滤歩骤中, 滤孔的目数为 100-200目。
另外, 上述本发明提供的组合物在制备具有如下任一一种功能产品中的应用, 也 属于本发明的保护范围-
1 ) 保湿;
2) 抗衰老;
3 ) 抗过敏;
4) 防皲裂。
本发明选用石斛、 三色堇、 麦冬、 燕麦原料、 银耳五种植物组合提取后外用, 通 过持水、维持正常水通道蛋白生理功能等途径,达到保湿作用同时兼具延缓皮肤衰老、 抗敏感及防皲裂的功效。
附图说明
图 1为皮肤表面的含水量百分比随时间的变化曲线。
说 明 书 图 2为皮肤表面的水份散失量随时间的变化曲线。
图 3为组合物对 DPPH的清除效果。
图 4为组合物对透明质酸酶的抑制效果。
具体实施方式
下面结合具体实施例对本发明作进一歩阐述, 但本发明并不限于以下实施例。 所 述方法如无特别说明均为常规方法。 所述原材料如无特别说明均能从公开商业途径而 得。 实施例 1、
按照如下歩骤制得中药组合物:
1 ) 将石斛、 麦冬、 银耳剪碎, 三色堇、 燕麦麸皮粉碎后, 按照质量配比石斛- 三色堇: 麦冬: 银耳: 燕麦麸皮: 水 =12: 6: 3: 2: 2: 160混匀后放置于电热恒温水 浴锅中, 于 80°C提取 2小时, 将提取结束的溶液从恒温水浴锅中取出, 在室温下静置 冷却 2h至常温后, 用 200目纱布进行粗过滤, 滤出药渣, 得滤液。在布氏漏斗里铺两 层滤纸,滤纸中间夹 0.5cm左右硅藻土,将粗过滤所得滤液进行真空抽滤,收集滤液, 得到本发明提供的中药组合物 A。 实施例 2、
按照如下歩骤制得中药组合物-
1 ) 将石斛、 麦冬、 银耳剪碎, 三色堇、 燕麦麸皮粉碎后, 按照配比石斛: 三色 堇- 麦冬: 银耳: 燕麦麸皮: 水 =5: 8: 2: 1: 1: 220, 混匀后放置于电热恒温水浴锅 中, 于 100°C提取 4小时, 将提取结束的溶液从恒温水浴锅中取出, 在室温下静置冷 却 4h至常温后, 用 100目纱布进行粗过滤, 滤出药渣, 得滤液。在布氏漏斗里铺两层 滤纸, 滤纸中间夹 0.5cm左右硅藻土, 将粗过滤所得滤液进行真空抽滤, 收集滤液, 得到本发明提供的中药组合物 B。 实施例 3、
按照如下步骤制得中药组合物-
1 ) 将石斛、 麦冬、 银耳剪碎, 三色堇、 燕麦麸皮粉碎后, 按照配比石斛: 三色 堇: 麦冬: 银耳: 燕麦麸皮: 水 =12: 3: 6: 5: 4: 200, 混匀后放置于电热恒温水浴 锅中, 于 65°C提取 1小时, 将提取结束的溶液从恒温水浴锅中取出, 在室温下静置冷 却 lh至常温后, 用 200目纱布进行粗过滤, 滤出药渣, 得滤液。在布氏漏斗里铺两层 滤纸, 滤纸中间夹 0.5cm左右硅藻土, 将粗过滤所得滤液进行真空抽滤, 收集滤液,
说 明 书 得到本发明提供的中药组合物 C。 实施例 4、 实施例 1、 实施例 2、 实施例 3所得中药组合物 A、 B、 C的保湿效果 评价
1.保湿效果——人体皮肤角质层水合率试验 /角质层水分散失试验
测试样品为膏霜, 均按照如下方法配制而得- 将实施例 1-3任一所得中药组合物加入到空白膏霜基质的 B相得到膏霜, 其中中 药组合物在膏霜中的质量百分含量均为 5%。
其中, 实施例 1所得中药组合物 A对应膏霜 1, 实施例 2所得中药组合物 B对应 膏霜 2, 实施例 3所得中药组合物 C对应膏霜 3。
另以所用空白膏霜基质作为空白对照。
所用空白膏霜基质的制备方法如下- 按照表 1所得配方, 将 A相各原料加热, 混合均匀, 加热到 82 ; 将 B相各原 料加热, 混合均匀, 加热到 82°C ; 在 B相中于 2000r/min的转速下均质, 将 A相缓慢 倒入 B相中, 均质 5min; 均质完成后, 搅拌降温; 40Ό时加入 C相, 搅拌均匀, 即 得空白膏霜基质。
表 1、 空白膏霜基质配方
测试前经专业人员培训过的健康受试者 30名 (男女各 15名), 年龄在 20~30岁,
说 明 书 所有受试者均无皮肤或系统性疾患史, 受试部位无异常, 且受试期间不涂抹任何与实 验无关的药物或者化妆品。
测试场所室温 (25±1 ) V , 相对湿度 (40±5 ) %, 测试前受试者应保持机体为稳 定状态。使用 35Ό左右的清水擦拭受试者双臂后, 于测试环境下静坐 30min后开始测 试。
在受试者手臂内侧距手掌基部 5cm处, 标记受测区域 (4Cmx4Cm), 同一手臂可 同时标记多个区域 (间隔 lcm) , 测试样品均随机分布。 先测量各测试区域的空白值, 然后测试样按 (2.0±0.1 ) mg/cm2的用量进行单次涂布, 涂布后釆用多功能皮肤水分 测试仪和皮肤水分散失仪分别测量 0.5h、 l h 、 2 h和 4 h受试区域和空白对照区域的 皮肤水合状态值、 水分散失量值。
测试将受试者涂抹同一样品区域内 5次测得的数据取平均值, 采用 SPSS统计软 件进行 t检验分析。
实验结果
①皮肤水分含量变化
水分含量变化反映在测试周期内, 实验区域水分含量随时间变化规律其值越大, 水分含量越大, 反之, 水分含量越小。
水合率 = (样品组水分含量一起始水分含量 )/起始水分含量
所得结果如图 1所示。
结果显示: 各实施例所得中药组合物均具有很好的抗干燥保湿作用, 其中以实施 例 1 中药组合物 A 的抗干燥保湿效果最好。
②皮肤水分散失变化
皮肤水分散失量 (Transepidermal water loss,TEWL) 反映了测试周期内, 实验区 域水分散失随时间的变化规律, 它可以表征测试样品的锁水功能, 其值越小, 水分散 失越少, 锁水能力越强; 反之, 锁水能力越弱。
失水率 (%H样品组水分散失量一起始水分散失量 )/起始水分散失量 X 100 水分散失结果见图 2。
结果显示: 4h 内, 各实施例所得中药组合物均具有锁水效果, 且中药组合物 A 的失水率最低, 锁水效果最显著。
综上所诉, 本发明提供的中药组合物能使皮肤水分含量升高, 水分散失降低, 具 有很好的锁水能力, 对皮肤具有显著的保湿效果。 实施例 5、实施例 1、实施例 2、实施例 3所得中药组合物 A、 B、 C对 AQP3 mRNA
说 明 书 表达量的影响
水通道蛋白 (aquaporins, AQPs)是一组与水、甘油及部分小分子物质跨膜转运相关 的细胞膜转运蛋白。 AQP3表达于包括皮肤在内的多种组织和器官。 正常表皮 KC仅 在基底细胞层表达 AQP3。 AQP3为上皮细胞膜提供一个孔道,保持细胞内渗透压和细 胞容积, 在水的转运中发挥作用。 AQP3 的甘油转运功能对皮肤的水合作用, 弹性保 持及损伤后的修复功能起着关键性作用。 本研究的组合物可通过保湿途径发挥修复皮 肤屏障和提高角质层水含量作用外, 可能还通过调节 AQP3水平而发挥这些作用, 我 们对 C3H鼠在应用中药组合物 A、 B、 C溶液后,采用实时定量 PCR检测鼠皮中 AQP3 mRNA的表达含量, 并与生理盐水做对照。
实验方法:
( 1 ) 分组: 实验小鼠共 20只, 分为实施例 1样品组 A、 实施例 2样品组 B、 实 施例 3样品组 C、 阴性对照组, 每组 5只。
样品组: 将实施例 1-3任一所得中药组合物和去离子水以 5: 95的质量比混匀而 得中药组合物的水溶液, 将该中药组合物的水溶液分别涂抹在实验小鼠上, 每天涂抹 1次;
对照组: 每天涂抹生理盐水 1次, 即保持正常生理状态的皮肤。
(2)先用脱毛膏将小鼠上腹部毛发去除, 按分组分别每天在每只小鼠腹部涂抹 1 次, 吸收后放入词养盒中, 30天后处死取皮肤。
(3 ) 皮肤用 dispase分离表皮, 通过胰酶分离成单个角质形成细胞。 用 Trizol抽 提角质形成细胞中的 RNA, 并逆转录成 cDNA后通过实时定量 PCR SYBRGREEN法 (QIAGEN 204143) 。 PCR 的 引 物 序 列 及 条 件 如 下 : aqp3 sense : 5'-TTGGTGGCTGGCCAAGTGTC-3';aqp3
antisense: 5' -GTCTGTGCC AGTGC ATAGAT-3 ' ; B— actin sense :
5'-TGTATGCCTCTGGTCGTACC-3';B— actin antisense :
5' -C AGGTCC AGACGC AGGATG-3 '。
94°C2 min; 94°C40 s; 62°C30 s; 72°C20 s; 35个循环。
(4)统计学方法结果用 One— way ANOVA t检验。
不同方法处理后 AQP3 mRNA表达量如表 2:
表 2、 不同添加量组合物溶液对小鼠皮肤 AQP3表达量的影响
] AQP3/B-actin
实施例 1样品组 A ~~ (5.004+0.015) E-03*
实施例 2样品组 B ( 4.08+0.05 ) E-03
说 明 书
实施例 3样品组 C (4.31+0.17) E-03
阴性对照组 (3.89±0.26) E-03
*表示与空白对照相比, P<0.05
实验结果显示,正常小鼠皮肤中,中药组合物有上调 AQP3 mRNA表达量的作用, 通过激发角质层水或甘油的运输, 增强皮肤水合作用, 皮肤中水分结合分子 (如透明 质酸) 的产生增强、 皮肤干燥脱落现象减轻。 实施例 6、 实施例 1、 实施例 2、 实施例 3所得中药组合物 A、 B、 C的抗衰老功 能评价
现代研究表明, 日光辐射能够引发皮肤光老化。 日光中的紫外辐射能够引发皮肤 氧化应激反应, 产生过量的自由基, 造成细胞损伤; 而红外辐射能够引起皮肤真皮细 胞中基质金属蛋白酶 -1 (MMP-1 ) 的高表达, 进而加速真皮层中弹性蛋白和胶原蛋白 的降解, 令肌肤弹性丧失、 纹理加深。 本专利中涉及的中药组合物具有清除自由基与 抑制 MMP-1的功效, 能够延缓因日光中紫外与红外辐射造成的皮肤光老化现象。
1 ) 自由基清除功效
DPPH (1, l-disphenyl-2-picryl-hydrazyl),化学命名为 1, 1二苯基 -2-三硝基苯肼, 是一种稳定的有机自由基。采用分光光度法进行定量分析实施例 1、实施例 2、实施例 3所得的中药组合物 A、 B、 C对自由基的清除情况。
将实施例 1-3任一所得中药组合物和去离子水以 5: 95的质量比混匀, 得到中药 组合物的水溶液;
以该中药组合物的水溶液作为试验组;
其中, 实施例 1所得中药组合物 A对应样品 1 ;
实施例 2所得中药组合物 B对应样品 2;
实施例 3所得中药组合物 C对应样品 3; 将试验组中的中药组合物替换为维生素 C衍生物, 所得水溶液作为阳性对照; 测定结果如图 3。
实验结果表明,各实施例所得中药组合物在其水溶液中的质量百分含量为 5.0%时, 均巳具有对 DPPH清除能力, 具有降低羟自由基、 烷自由基或氧化自由基的有效浓度 和打断脂质过氧化链反应的作用, 从而具有良好的延缓皮肤光老化的作用。
2) 基质金属蛋白酶 -1抑制作用
采用荧光底物法检测中药组合物对 MMP-1活性的影响。 实验方法参照 《两种化
说 明 书 妆品延缓衰老功效添加剂体外筛选方法的建立及应用研究》(来吉祥, 2007)所述实验 方法。
将实施例 1-3任一所得中药组合物和去离子水以 5: 95的质量比混匀, 得到中药 组合物的水溶液;
对上述水溶液进行测定, 所得结果如表 3 ;
表 3、 组合物对 MMP-1抑制效果
名称 抑制率 (%)
1 实施例 1组合物 A 81.63
2 实施例 2组合物 B 47.64
3 实施例 3组合物 C 60.98 结果显示, 各实施例组合物, 均具有对由红外辐射诱导产生的 MMP-1 的抑制作 用, 能够减少由于光照引起的胶原蛋白与弹性蛋白的过度降解, 保持皮肤弹性, 减轻 皮肤光老化的作用。 实施例 7、 实施例 1、 实施例 2、 实施例 3所得中药组合物 A、 B、 C的抗敏功效 评价
透明质酸是组织基质中具有限制水分及其它细胞外物质扩散作用的成分, 经透明 质酸酶水解后, 细胞间则成为非粘性, 细胞脱颗粒和合成的新介质渗出, 发挥生物效 应, 导致速发型过敏反应的发生。 因此常用样品对透明质酸酶的抑制程度来反应样品 对 I型超敏反应的缓解和改善作用。
按照下述方法测定实施例 1、 实施例 2、 实施例 3所得中药组合物 、 B、 C对透 明质酸酶的抑制程度- 将实施例 1所得中药组合物 A和去离子水以 5: 95的质量比混匀得到样品溶液
C1 ;
将实施例 2所得中药组合物 B和去离子水以 5: 95的质量比混匀得到样品溶液
C2;
将实施例 3所得中药组合物 C和去离子水以 5: 95的质量比混匀得到样品溶液
C3;
试验设三次重复。 每次重复的处理方法如下:
取 7个试管, 分别标记为 A、 B、 Cl-C3、 D和 E;
按照表 4各对应列的顺序添加试剂, 并进行相应步骤的操作:
说 明 书 其中, A管添加的为对照溶液, 对应表 4中 A列;
B管添加的为对照空白溶液, 对应表 4中 B列;
D管添加的为样品空白溶液, 对应表 4中 D列;
E管添加的为阳性对照溶液, 对应表 4中 E列;
C1-C3管均按照表 4中 C列的顺序添加试剂;
C1管添加的样品溶液为样品溶液 C1 ;
C2管添加的样品溶液为样品溶液 C2;
C3管添加的样品溶液为样品溶液 C3;
测定上述各管在波长为 555nm处的吸光度值, 也即 ABS值后, 按照如下抗过敏 活性计算公式计算样品溶液对透明质酸酶的抑制率- 样品的透明质酸酶抑制率 =[ (A-B ) -(C-D)]/(A-B)xlOO%
阳性对照的透明质酸酶抑制率=[ (A-B ) -(E-D)]/(A-B)xlOO%
上述两公式中, A均为 A管的 ABS值;
B为 B管的 ABS值;
C为 C管的 ABS值 ;
D为 D管的 ABS值;
E为 E管的 ABS值; 其中, C管的 ABS值的测定方法如下-
1 ) 取 0.1 mL0.25 mmol/L CaCl2溶液和 0.5mL透明质酸酶液 37°C水浴保温培养 20min;
2) 再加入处理后的样品溶液 0.5 mL, 继续 37°C水浴保温培养 20 min;
3 ) 再加入 0.5 mL透明质酸钠水溶液 37°C水浴保温 30 min, 常温放置 5 min;
4) 再加入 0.1 mL0.4 mol/L NaOH水溶液和 0.5 mL乙酰丙酮, 置于沸水浴中加热 15 min后立即用冰水进行冷却 5 min;
5 )再加入埃尔利希试剂 l.O mL并用 3.0 mL无水乙醇进行稀释, 常温放置 20 min 显色, 用分光光度计测定其吸光度值, 所得结果即为 C——样品溶液的 ABS值; 上述 A组至 E组的试验步骤亦见于表 4中。
表 4、 透明质酸酶抑制实验操作表
说 明 书
注: "+"为此项添加, "- "为此项不添加, 但用等体积的醋酸缓冲溶液代替; " * " 为用等体积的质量百分浓度为 5%的甘草酸二钾的水溶液代替。
所得结果如图 4所示。
其中, 实施例 1所得组合物 A的透明质酸酶抑制率为 75.32% ;
实施例 2所得组合物 B的透明质酸酶抑制率为 59.89% ;
实施例 3所得组合物 C的透明质酸酶抑制率为 67.26%。
E管也即阳性对照的透明质酸酶抑制率为 92.12%。
实验结果表明, 各实施例组合物, 均具有体外抑制透明质酸酶功效, 能够抑制过 敏介质的释放, 从而具有良好的抗过敏的作用。 实施例 8、 实施例 1、 实施例 2、 实施例 3所得中药组合物 A、 B、 C的防皲裂效 果
在表 5所示空白膏霜基质配方 B相中分别加入 3%的中药组合物 A混匀, 即得实 施例 1组合物膏霜。
按照与上相同的方法,仅将中药组合物 A替换为实施例 2所得中药组合物 B或实 施例 3所得中药组合物 C , 依次得到实施例 2组合物膏霜、 实施例 3组合物膏霜。
说 明 书 将上述三种膏霜进行人体足部皮肤涂抹试验, 时间为一个月 (冬季),试用人数 120 人, 每组 30人, 随机分为 4组, 分别为 A组、 B组、 C组、 空白对照组(空白膏霜基 质)。
其中, A组涂抹实施例 1组合物膏霜;
B组涂抹实施例 2组合物膏霜;
C组涂抹实施例 3组合物膏霜; 所用空白膏霜基质的制备方法如下:
按照表 5所得配方, 将 A相各原料加热, 混合均匀, 加热到 82°C ; 将 B相各原 料加热, 混合均匀, 加热到 82°C ; 在 B相中于 2000r/min的转速下均质, 将 A相缓慢 倒入 B相中, 均质 5min; 均质完成后, 搅拌降温; 40Ό时加入 C相, 搅拌均匀, 即 得空白膏霜基质。
表 5、 空白膏霜基质配方
使用者用后反映该产品不仅对皮肤有营养滋润作用, 且对皮肤的干裂伤口有良好 的愈合修复作用, 试用结果如表 6:
表 6 对中药组合物防皲裂效果人体应用评估表
粗糙改善程度 湿润度 柔软性
明显 一般 无 明显 一般 无 明显 一般 无 实施例 1组合物 A 21 7 2 23 7 0 22 7 1 实施例 2组合物 B 15 12 3 25 5 0 18 10 2 实施例 3组合物 C 17 11 2 25 5 0 20 10 0 空白对照 11 15 4 20 9 1 17 7 6
说 明 书 调査结果证实中药组合物有如下特点:
1.产品中所含中草药有效成分能使其增强机体自卫能力, 刺激肉芽组织生长。
2.可在局部形成保护膜、 缩短凝血时间, 减少出血, 从而有利于创面的愈合。
3.能有效地保持肌肤的水分而无甘油类护肤品的油腻感, 使用之后柔润舒适。 产品中所含中草药有效成分能使皮肤增强机体自卫能力, 可在局部形成保护膜、 减少出血, 从而有利于创面的愈合, 使用者反映良好。
Claims
1、 一种中药组合物, 其活性成分为如下原料混合物的水提液- 石斛、 三色堇、 麦冬、 银耳和燕麦原料;
其中, 所述燕麦原料为燕麦粒、 燕麦粉或燕麦麸皮。
2、 根据权利要求 1 所述的组合物, 其特征在于: 所述石斛、 三色堇、 麦冬、 银 耳和燕麦原料的质量比为 5-15: 2-10: 2-10: 1-5: 1-5。
3、 根据权利要求 1或 2所述的组合物, 其特征在于: 所述水提液中, 水提步骤 所用水与石斛的质量比为 160-220: 5-15。
4、根据权利要求 1-3任一所述的组合物, 其特征在于: 所述水提液为按照包括如 下步骤的方法制备而得:
将所述石斛、 三色堇、 麦冬、 银耳、 燕麦原料和水按配比混匀后提取而得; 所述提取歩骤中, 温度为 60-100°C ; 时间为 1-4小时。
5、 一种制备权利要求 1-4任一所述组合物的方法, 包括如下步骤:
将所述石斛、 三色堇、 麦冬、 银耳、 燕麦原料和水按照权利要求 2或 3所述配比 混匀后于提取得到水提液。
6、根据权利要求 5所述的方法,其特征在于:所述提取步骤中,温度为 60-100°C; 时间为 1-4小时。
7、 根据权利要求 5或 6所述的方法, 其特征在于: 所述方法还包括如下步骤: 在所述提取歩骤之后, 将所得水提液静置后过滤, 收集滤液。
8、 根据权利要求 7所述的方法, 其特征在于: 所述静置歩骤中, 温度为室温, 时间为 1-4小时;
过滤歩骤中, 滤孔的目数为 100-200目。
9、 权利要求 1-6任一所述组合物在制备具有如下任一一种功能产品中的应用:
1 ) 保湿;
2) 抗衰老;
3 ) 抗过敏;
4) 防皲裂。
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