WO2024031762A1 - 一种护肤组合物和制备方法及其用途 - Google Patents

一种护肤组合物和制备方法及其用途 Download PDF

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WO2024031762A1
WO2024031762A1 PCT/CN2022/116848 CN2022116848W WO2024031762A1 WO 2024031762 A1 WO2024031762 A1 WO 2024031762A1 CN 2022116848 W CN2022116848 W CN 2022116848W WO 2024031762 A1 WO2024031762 A1 WO 2024031762A1
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parts
preparation
skin
care composition
skin care
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English (en)
French (fr)
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王莲婧
赵玲玲
张华�
姚昀
梁绮莹
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健合香港有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/01—Instant products; Powders; Flakes; Granules
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/15—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/455—Nicotinic acids, e.g. niacin; Derivatives thereof, e.g. esters, amides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/728—Hyaluronic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/75—Rutaceae (Rue family)
    • A61K36/752—Citrus, e.g. lime, orange or lemon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/31—Extraction of the material involving untreated material, e.g. fruit juice or sap obtained from fresh plants
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to the field of medical technology, and in particular to a skin care composition, a preparation method and its use.
  • Aging is a direct manifestation of the slowdown of human body functions and is also a natural process.
  • the main causes of aging are excessive oxidation of the human body, reduction of cell lifespan, aging of proteins, and microcirculation disorders.
  • As people age, their metabolism slows down, resulting in slower cell metabolism and slower growth.
  • the skin cannot replenish nutrients in time, leading to skin aging.
  • Another example is the increase in natural environmental pollution and atmospheric damage, which causes an increase in ultraviolet rays irradiating the ground, causing skin aging.
  • Whitening mainly works by inhibiting the activity of tyrosinase in the skin, which can significantly reduce the content of melanin in cells, thereby achieving the whitening effect.
  • Hydrating and moisturizing refers to preventing the evaporation of skin water, moisturizing the skin, improving microcirculation and enhancing skin moisture.
  • the present invention provides a composition with skin care and anti-aging effects, and has broad market prospects in the preparation of medicines and health foods.
  • the technical problem to be solved by the present invention is to provide a skin care composition, a preparation method and its use.
  • the skin care composition has skin care and anti-aging effects.
  • a skin care composition consisting of the following components by weight:
  • Vitamin C in the present invention is L-ascorbic acid.
  • the blood orange powder is an extract of Tarocco, Moro and/or Sanginello blood oranges.
  • the preparation method of blood orange powder includes: taking blood oranges and cold pressing at low temperature to obtain blood orange juice;
  • the blood orange juice is filtered, concentrated, a carrier is added, and dried;
  • the low-temperature cold pressing temperature is 0-5°C;
  • the concentration step low-temperature vacuum drying is used for concentration, the temperature is 35-45°C, and the vacuum degree is 0.095-0.1MPa;
  • the carrier includes: maltodextrin, inulin or resistant dextrin; the mass percentage of the carrier in the blood orange powder is 40-60%;
  • the drying is spray drying.
  • the collagen peptide is extracted from cowhide or pigskin
  • the average molecular weight of the collagen peptide is less than 2000 Da.
  • a method for preparing a skin care composition weighing the raw materials of each component according to the formula and mixing; or
  • the temperature of the water is 70-90°C;
  • the stirring condition is a rotation speed of 80-110 r/min, stir for 240-360 seconds until completely dissolved, and continue stirring at a rotation speed of 40-50 r/min for 20-50 minutes.
  • the sterilization condition is high-temperature instant sterilization at 110-123°C for 5-10 seconds.
  • a preparation using the skin care composition or the skin care composition prepared by the method as an active ingredient is provided.
  • excipients or carriers permitted by the formulation are also included.
  • the form of the preparation includes liquid preparation and solid preparation;
  • the preparation forms include injections, tablets, capsules, granules, powders or ointments.
  • the skin care composition, the skin care composition prepared by the method or the preparation has any of the following uses:
  • col1a1a gene i. Use in the preparation of drugs or foods that regulate the expression level of col1a1a gene and/or col1a1b gene.
  • a skin care composition consisting of the following components by weight: 1-50 parts of blood orange powder, 10-1000 parts of collagen peptide, and 0.1-50 parts of vitamin C; the above-mentioned skin care composition , blood orange powder, collagen peptides and vitamin C synergistically play a role in skin care. It has been experimentally proven to effectively increase skin moisture, reduce skin melanin, reduce skin fine lines, and has the effects of whitening, hydrating and delaying aging.
  • a skin care composition consisting of the following components by weight: 1-50 parts of blood orange powder, 10-1000 parts of collagen peptide, 0.1-50 parts of vitamin C, 0.1-50 parts of sodium hyaluronate 50 parts, and 0.01-20 parts of nicotinamide; the above-mentioned skin care composition exerts a skin care effect through the synergy between each component, and has significant whitening, hydrating and anti-aging effects.
  • the blood orange powder in the skin care composition of the present invention is preferably from unique varieties of blood oranges on Sicily, Italy, including Tarocco, Moro and Sanguinello.
  • the blood orange powder is rich in anthocyanins, flavonoids, and vitamin C.
  • Anthocyanins have extremely strong antioxidant capabilities and can directly scavenge free radicals produced by human body metabolism and exert antioxidant effects.
  • the vitamin C content is 99wt% or more
  • the sodium hyaluronate content is 95wt% or more
  • the citric acid content is 99wt% or more
  • the nicotinamide content is 99wt% or more.
  • the collagen peptides in the following examples are extracted from pig skin, have an average molecular weight of less than 2000 Da, and are commercially available products.
  • the blood orange powder in the following examples is prepared by the following preparation method: take blood oranges from Tarocco, Moro and/or Sanginello (blood oranges from Tarocco are selected in the present invention), and process them at low temperature ( 0-5°C) to obtain blood orange juice, then filter, use low-temperature vacuum drying for concentration, temperature 35-45°C, vacuum degree 0.095-0.1MPa, and then add carriers (maltodextrin, inulin or resistant dextrin , in the present invention, maltodextrin is selected, and the carrier accounts for 50% of the mass percentage of the finally prepared blood orange powder), and then spray-dried.
  • a preparation method take blood oranges from Tarocco, Moro and/or Sanginello (blood oranges from Tarocco are selected in the present invention), and process them at low temperature ( 0-5°C) to obtain blood orange juice, then filter, use low-temperature vacuum drying for concentration, temperature 35-45°C, vacuum degree 0.095-0.1MPa, and then add carriers (mal
  • Example 1 Group blood orange (kg) Collagen peptide(kg) Vitamin C (kg) Sodium hyaluronate Nicotinamide
  • Example 1 1 1000 0.1 (k/) (k/)
  • Example 2 50 10 50 / / Example 3 25 500 50 / /
  • Example 4 1 1000 0.1 0.1 10
  • Example 5 50 10 50 50 0.01
  • Example 6 25 500 50 25 20
  • Packaging Install the aluminum foil film on the packaging machine. After debugging is completed, unload the materials for packaging. The net weight should be controlled at 3 grams per bag.
  • the samples are all prepared into 2.00 mg/mL mother liquor with standard dilution water and are ready for use.
  • the above-mentioned samples include: the skin care compositions prepared in Examples 1-6 (see Table 1), and the control group includes nicotinamide (positive control), vitamin C, collagen peptide, sodium hyaluronate, and the independent raw materials in the formula of the present invention. Blood orange powder.
  • Zebrafish were all raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt added to each 1L of reverse osmosis water, conductivity of 450 to 550 ⁇ S/cm; pH of 6.5 to 8.5; hardness of 50 to 100 mg/L CaCO 3 )
  • the experimental animal use license number is: SYXK (Zhejiang) 2012-0171. Feeding and management comply with the requirements of international AAALAC certification (certification number: 001458).
  • Wild-type AB strain zebrafish is reproduced through natural pair mating. Zebrafish aged 6 hours after fertilization (6hpf) were used for MTC determination of whitening efficacy and its efficacy evaluation.
  • Pierce TM Coomassie (Bradford) protein detection kit (lot number WE317426, Thermo Fisher Scientific (China) Co., Ltd., the United States); RIPA lysis buffer (lot number 0205212120315, Shanghai Beyotime Biotechnology Co., Ltd., China); PMSF (Batch No. ST505, Shanghai Biyuntian Biotechnology Co., Ltd., China); Levodopa (Batch No. I170522, Shanghai Aladdin Biochemical Technology Co., Ltd., China).
  • Examples 1-6 all have whitening effect, which is specifically manifested by inhibiting tyrosinase activity. See Table 3 for details.
  • the samples are all prepared into 2.00 mg/mL mother liquor with standard dilution water and are ready for use.
  • Collagen peptide uses Verisol collagen, white powder, batch number 893660, GELITA, store in a cool place and away from light. Use standard dilution water to prepare a 2.00mg/mL stock solution and prepare it for immediate use. Niacinamide, vitamin C, sodium hyaluronate, blood orange powder.
  • Zebrafish were all raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt added to each 1L of reverse osmosis water, conductivity of 450 to 550 ⁇ S/cm; pH of 6.5 to 8.5; hardness of 50 to 100 mg/L CaCO 3 )
  • the experimental animal use license number is: SYXK (Zhejiang) 2012-0171. Feeding and management comply with the requirements of international AAALAC certification (certification number: 001458).
  • Wild-type AB strain zebrafish is reproduced through natural pair mating. Zebrafish aged 4 dpf were used for evaluation of anti-wrinkle and firming efficacy.
  • Absolute ethanol (batch number 20210107, Sinopharm Chemical Reagent Co., Ltd., China); FastQuant RT Kit (With gDNase) kit (catalog number KR106, TIANGEN, China); RNA-Quick Purification Kit (RNA rapid extraction kit) (catalog number RN001 , YiShan Biotech, China); iTaq Universal SYBR Green Supermix (Cat. No. 1725124, Bio-rad, USA).
  • RNA Take 2.00 ⁇ g of total RNA from the zebrafish sample, follow the instructions of the cDNA first-strand synthesis kit, synthesize 20.0 ⁇ L cDNA, and detect the expression of ⁇ -actin, col1a1a, and col1a1b genes by q-PCR.
  • ⁇ -actin was used as the internal reference for gene expression to calculate the relative RNA expression of col1a1a and col1a1b genes.
  • Statistical processing results are expressed as mean ⁇ SE.
  • Statistical analysis was performed using SPSS26.0 software, and p ⁇ 0.05 indicated that the difference was statistically significant. 3.Test results
  • Examples 1-6 have better effects than individual vitamin C, collagen peptides and blood orange powder, and have significant differences. Therefore, the groups causing significant differences were compared with Examples 1-6, and different letters indicate significant differences (P ⁇ 0.05). It is proved that although the formula only has a lower concentration of collagen peptides, it can show more significant effects than high-concentration collagen peptides.
  • This experimental example examines the efficacy of the skin care composition of the present invention in whitening and anti-aging.
  • Example 7 Take 3g every day
  • Example 8 Take 30mL every day
  • the number of volunteers in each of the above embodiment groups was 30. Take it continuously for 8 weeks, and conduct relevant tests at weeks 0, 4, and 8 to analyze the changes before and after.
  • Test parts and test items cheek moisture, forearm moisture, skin elasticity in crow’s feet, and melanin content in the whole face.
  • Test moisture use (C+K, Germany), measure the moisture of both cheeks and forearms, and take the average of three measurements;
  • Test method for skin elasticity of crow's feet Use MPA580 (C+K, Germany) to measure the skin elasticity of crow's feet, and take the average of three measurements;
  • the experimental data were subjected to repeated measures analysis of variance test and generalized estimating equation (GEE) regression analysis on week 0 (baseline), fourth week, and eighth week using SPSS 21.0.
  • GOE generalized estimating equation
  • taking the skin care composition of the present invention can significantly increase the moisture of the cheeks, the moisture of the forearms, and the skin elasticity of the crow's feet, significantly reduce the melanin content of the whole face, and achieve better whitening, hydrating, and anti-wrinkle effects. Remarkable effect.

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Abstract

涉及医药技术领域，具体涉及一种护肤组合物和制备方法及其用途，包括如下重量份的组分组成：血橙粉1-50份，胶原蛋白肽10-1000份，和维生素C 0.1-50份；血橙粉、胶原蛋白肽和维生素C协同发挥护肤的作用，经过实验验证，该组合物可以高效地增加皮肤水分、减少皮肤黑色素、减少皮肤细纹，具有美白、补水和延缓衰老的作用。

Description

一种护肤组合物和制备方法及其用途 技术领域
本发明涉及医药技术领域，具体涉及一种护肤组合物和制备方法及其用途。
背景技术
衰老是人体机能变缓的直接表现，也是一种自然过程，衰老的主要原因为人体的过度氧化、细胞寿命的降低、蛋白质的老化和微循环障碍等。比如随着年龄的增长，人的新陈代谢会变慢，进而细胞代谢较慢、生长速度慢，不能及时补充皮肤的营养，导致皮肤老化。再如大自然环境污染和大气层破坏程度的增大使得照射到地面的紫外线不断增加，引起皮肤衰老。
美白主要是通过抑制肌肤中络氨酸酶活性，能明显减少细胞内黑色素的含量，从而达到美白效果。
补水保湿是指防止肌肤水份的蒸发，滋润肌肤、改善微循环和增强肌肤湿润度等方面。
为改善人的皮肤情况，本发明提供一种具有护肤、抗衰老的作用的组合物，在制备药物、保健食品方面具有广阔的市场前景。
发明内容
因此，本发明要解决的技术问题在于提供一种护肤组合物和制备方法及其用途，所述护肤组合物具有护肤、抗衰老的作用。
为此，本发明提供了如下的技术方案：
一种护肤组合物，由如下重量份的组分组成：
血橙粉1-50份，胶原蛋白肽10-1000份，和维生素C 0.1-50份。
可选的，由如下重量份的组分组成：
血橙粉1份，胶原蛋白肽1000份，和维生素C 0.1份；或
血橙粉50份，胶原蛋白肽10份，和维生素C 50份；或
血橙粉25份，胶原蛋白肽500份，和维生素C 50份。
可选的，由如下重量份的组分组成：
血橙粉1-50份，胶原蛋白肽10-1000份，维生素C 0.1-50份，透明质酸钠0.1-50份，和烟酰胺0.01-20份；
可选的，由如下重量份的组分组成：
血橙粉1份，胶原蛋白肽1000份，和维生素C 0.1份，透明质酸钠0.1份，烟酰胺10份；或
血橙粉50份，胶原蛋白肽10份，和维生素C 50份，透明质酸钠50份，烟酰胺0.01份；或
血橙粉25份，胶原蛋白肽500份，和维生素C 50份，透明质酸钠25份，烟酰胺20份。
本发明中的维生素C为L-抗坏血酸。
可选的，所述的血橙粉为塔罗科、莫罗和/或桑吉耐洛的血橙的提取物。
可选的，所述血橙粉的制备方法，包括：取血橙，低温冷榨，得到血橙汁；
将所述血橙汁过滤，浓缩，添加载体，干燥；
可选的，所述低温冷榨温度为0-5℃；
可选的，所述浓缩步骤中采用低温真空干燥进行浓缩，温度35～45℃，真空度0.095～0.1MPa；
可选的，所述载体包括：麦芽糊精、菊粉或抗性糊精；所述载体占血橙粉的质量百分比为40-60％；
可选的，所述干燥为喷雾干燥。
可选的，所述胶原蛋白肽为由牛皮或猪皮中提取得到的；
可选的，所述胶原蛋白肽的平均分子量小于2000Da。
一种护肤组合物的制备方法，按照配方称取各组分的原料，混合；或
按照配方称取各组分的原料，加入水中，溶解，搅拌，灭菌；
可选的，所述水的温度为70-90℃；
可选的，所述搅拌条件为80-110r/min的转速，搅拌240～360秒，直至完全溶解，以40-50r/min的转速继续搅拌20-50分钟。
可选的，所述灭菌条件为110～123℃条件下高温瞬时灭菌5～10s。
一种制剂，以所述的护肤组合物或所述的方法制备得到的护肤组合物为活性成分。
可选的，还包括制剂允许的赋形剂或载体。
可选的，所述制剂的形式包括液体制剂和固体制剂；
可选的，所述制剂的形式包括注射剂、片剂、胶囊剂、颗粒剂、粉剂或膏剂。
所述的护肤组合物、所述的方法制备的护肤组合物或所述的制剂具有如下任意一项的用途：
a.在制备具有护肤作用的药物或食品中的用途；或
b.在制备具有美白皮肤作用的药物或食品中的用途；或
c.在制备具有皮肤补水作用的药物或食品中的用途；或
d.在制备具有延缓衰老作用的药物或食品中的用途；或
e.在制备具有减少皮肤黑色素作用的药物或食品中的用途；或
f.在制备具有减少皮肤细纹作用的药物或食品中的用途；或
f.在制备具有减少皮肤细纹、紧致皮肤和/或提高皮肤弹性作用的药物或食品中的用途；或
h.在制备抑制酪氨酸酶活性的药物或食品中的用途；或
i.在制备调控col1a1a基因和/或col1a1b基因的表达水平的药物或食品中的用途。
本发明技术方案，具有如下优点：
1.本发明提供的一种护肤组合物，由如下重量份的组分组成：血橙粉1-50份，胶原蛋白肽10-1000份，和维生素C 0.1-50份；上述的护肤组合物，由血橙粉、胶原蛋白肽和维生素C协同发挥护肤的作用，经过实验验证可以高效地内增加皮肤水分、减少皮肤黑色素、减少皮肤细纹，具有美白、补水和延缓衰老的作用。
2.本发明提供的一种护肤组合物，由如下重量份的组分组成：血橙粉1-50份，胶原蛋白肽10-1000份，维生素C 0.1-50份，透明质酸钠0.1-50份，和烟酰胺0.01-20份；上述的护肤组合物通过各组分之间协同发挥护肤的作用，具有显著的美白、补水和延缓衰老的作用。
3.本发明提供的一种护肤组合物，本发明所述护肤组合物中的血橙粉优选自意大利西西里岛上的特有品种的血橙，包括塔罗科、莫罗和桑吉耐洛。所述血橙粉富含花色苷、黄酮类、维生素C成分，花色苷具有极强的抗氧化能力，能直接清除人体代谢产生的自由基，发挥抗氧化作用。
具体实施方式
提供下述实施例是为了更好地进一步理解本发明，并不局限于所述最佳实施方式，不对本发明的内容和保护范围构成限制，任何人在本发明的启示下或是将本发明与其他 现有技术的特征进行组合而得出的任何与本发明相同或相近似的产品，均落在本发明的保护范围之内。
实施例中未注明具体实验步骤或条件者，按照本领域内的文献所描述的常规实验步骤的操作或条件即可进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规试剂产品。
下述实施例中维生素C含量99wt％以上、透明质酸钠含量95wt％以上、柠檬酸含量99wt％以上、烟酰胺含量99wt％以上。
下述实施例中的胶原蛋白肽为从猪皮中提取得到，平均分子量小于2000Da，为市售产品。
下述实施例中的血橙粉，采用如下制备方法制备：取塔罗科、莫罗和/或桑吉耐洛的血橙(在本发明中选择塔罗科的血橙)，经低温(0-5℃)冷榨，得到血橙汁，然后过滤，采用低温真空干燥进行浓缩，温度35～45℃，真空度0.095～0.1MPa，然后添加载体(麦芽糊精、菊粉或抗性糊精，本发明中选择麦芽糊精，载体占最终制备的血橙粉质量百分比50％)后，喷雾干燥。
实施例1-实施例6的护肤组合物的配方见下表1。
表1.护肤组合物实施例与对照组
组别 血橙(kg) 胶原蛋白肽(kg) 维生素C(kg) 透明质酸钠 烟酰胺
实施例1 1 1000 0.1 (k/) (k/)
实施例2 50 10 50 / /
实施例3 25 500 50 / /
实施例4 1 1000 0.1 0.1 10
实施例5 50 10 50 50 0.01
实施例6 25 500 50 25 20
实施例1-实施例6的护肤组合物的制备方法：按照配方称取原料，混合均匀，即得。实施例7本发明护肤组合物粉剂制剂的制备
1)称量、备料：称取实施例3所述组合物的原料、抗性糊精10kg、甜菊糖苷3kg、柠檬酸1kg备用。
2)混合：将上述原料混合25分钟。
3)包装：将铝箔膜安装在包装机上，待调试完毕后下料进行包装，净重控制在3克每袋。
实施例8本发明护肤组合物液体制剂的制备
1)称量、备料：称取实施例6所述组合物的原料、甜菊糖苷1kg、柠檬酸5kg称量备用。
2)混合：将上述原料分别溶解于温水中(80℃)，100r/min的转速，搅拌300秒，直至完全溶解，以50r/min的转速继续搅拌30分钟。121℃条件下高温瞬时灭菌5s，即得。
3)灌装：灭菌后液体在洁净区内直接灌装进包装瓶中。
实验例1美白功效
1.检测材料
1.1.样品配制信息
样品均用标准稀释水配制成2.00mg/mL母液，现配现用。
上述样品包括：实施例1-6制备的护肤组合物(见表1)、对照组包括本发明组方中单独的原料烟酰胺(阳性对照)、维生素C、胶原蛋白肽、透明质酸钠、血橙粉。
1.2.实验动物
斑马鱼均饲养于28℃的养鱼用水中(水质：每1L反渗透水中加入200mg速溶海盐，电导率为450～550μS/cm；pH为6.5～8.5；硬度为50～100mg/L CaCO 3)，实验动物使用许可证号为：SYXK(浙)2012-0171。饲养管理符合国际AAALAC认证(认证编号：001458)的要求。
野生型AB品系斑马鱼，以自然成对交配繁殖方式进行。年龄为受精后6小时(6hpf)的斑马鱼用于美白功效MTC测定及其功效评价。
1.3.仪器、耗材与试剂
解剖显微镜(SZX7，OLYMPUS，日本)；CCD相机(VertA1，上海土森视觉科技有限公司，中国)；精密电子天平(CP214，OHAUS，美国)；6孔板(Nest Biotech，中国)；高速冷冻离心机(Heraeus Fresco17，ThermoFisher，德国)；多功能酶标仪(SPARK，TECAN，瑞士)。
Pierce TM考马斯(Bradford)蛋白检测试剂盒(批号WE317426，赛默飞世尔科技(中国)有限公司，美国)；RIPA裂解液(批号0205212120315，上海碧云天生物技术 有限公司，中国)；PMSF(批号ST505，上海碧云天生物技术有限公司，中国)；左旋多巴(批号I170522，上海阿拉丁生化科技股份有限公司，中国)。
2.检测方法
2.1.MTC测定
随机选取6hpf野生型AB品系斑马鱼于6孔板中，每孔30尾。分别水溶给予样品组(终浓度见表2)，同时设置正常对照组，每孔容量为3mL。每天换液。28℃
处理4天后，确定各样品对正常斑马鱼的MTC。
2.2美白功效评价
随机选取6hpf野生型AB品系斑马鱼于6孔板中，每孔30尾。分别水溶给予样品(终浓度见表3)，阳性对照如胶原蛋白肽和烟酰胺1000μg/mL浓度，同时设置正常对照组，每孔容量为3mL。每天换液。平行设置三次实验。28℃处理4天后，采用多巴胺氧化法，在475nm波长处测定吸光度值，以该指标的统计学分析结果评价样品对斑马鱼酪氨酸酶活性的抑制功效。统计学处理结果采用mean±SE表示。用SPSS26.0软件进行统计学分析，p<0.05表明差异具有统计学意义。
3.检测结果
3.1.MTC
详见表2。
表2.美白功效浓度摸索实验结果(n＝30)
Figure PCTCN2022116848-appb-000001
Figure PCTCN2022116848-appb-000002
3.2.美白功效评价
在本实验条件下，实施例1-6均具有美白功效，具体表现为抑制酪氨酸酶活性。详见表3。
表3.美白功效实验结果(n＝3)
Figure PCTCN2022116848-appb-000003
与正常对照组比较，*p<0.05，**p<0.01，***p<0.001
由上述表3可知，在提高美白方面，与正常对照组比较，实施例1-6、维生素C、烟酰胺具有显著性差异。证明组方虽然只有较低浓度的维生素C或烟酰胺，但可以表现同高浓度维生素C和烟酰胺相当甚至更显著的效果。
实验例2抗皱紧致功效(I型胶原蛋白基因)
1.检测材料
1.1.样品配制信息
样品均用标准稀释水配制成2.00mg/mL母液，现配现用。
阳性对照：胶原蛋白肽采用Verisol胶原蛋白，白色粉末，批号893660，嘉利达，阴凉避光储存。用标准稀释水配制成2.00mg/mL母液，现配现用。烟酰胺、维生素C、透明质酸钠、血橙粉。
1.2.实验动物
斑马鱼均饲养于28℃的养鱼用水中(水质：每1L反渗透水中加入200mg速溶海盐，电导率为450～550μS/cm；pH为6.5～8.5；硬度为50～100mg/L CaCO 3)，实验动物使用许可证号为：SYXK(浙)2012-0171。饲养管理符合国际AAALAC认证(认证编号：001458)的要求。
野生型AB品系斑马鱼，以自然成对交配繁殖方式进行。年龄为4dpf的斑马鱼用于抗皱紧致功效评价。
1.3.仪器、耗材与试剂
解剖显微镜(SZX7，OLYMPUS，日本)；CCD相机(VertA1，上海土森视觉科技有限公司，中国)；精密电子天平(CP214，OHAUS，美国)；6孔板(Nest Biotech，中国)；普通PCR扩增仪(T100，BIO-RAD，新加坡)；荧光定量PCR仪(CFX Connect，BIO-RAD，新加坡)；高速冷冻离心机(Heraeus Fresco17，ThermoFisher，德国)；紫外-可见光分光光度计(Nanodrop2000，Thermo，奥地利)；微孔板迷你离心机(BE-6100，海门市其林贝尔仪器制造有限公司，中国)；低位裙边96孔板(透明)(HSP9601，Bio-rad，美国)；光学粘性封膜B(MSB1001，Bio-rad，美国)。
无水乙醇(批号20210107，国药集团化学试剂有限公司，中国)；FastQuant RT Kit(With gDNase)试剂盒(货号KR106，TIANGEN，中国)；RNA-Quick Purification Kit(RNA快速提取试剂盒)(货号RN001，YiShan Biotech，中国)；iTaq Universal SYBR Green Supermix(货号1725124，Bio-rad，美国)。
2.检测方法
随机选取4dpf野生型AB品系斑马鱼于6孔板中，每孔30尾。分别水溶给予样品 (终浓度见表4)，如阳性对照Verisol胶原蛋白肽250μg/mL浓度，同时设置正常对照组，每孔容量为3mL。平行设置三次实验。28℃处理24h后，使用RNA快速提取试剂盒提取各组斑马鱼总RNA，利用紫外-可见光分光光度计对总RNA浓度和纯度进行测定。取2.00μg斑马鱼样品总RNA，按照cDNA第一链合成试剂盒说明操作，合成20.0μL cDNA，通过q-PCR检测β-actin、col1a1a和col1a1b基因的表达。用β-actin作为基因表达的内参，计算col1a1a和col1a1b基因的RNA相对表达量。统计学处理结果采用mean±SE表示。用SPSS26.0软件进行统计学分析，p<0.05表明差异具有统计学意义。3.检测结果
3.1RNA提取结果及引物序列信息
在实验终点，提取斑马鱼总RNA，用紫外-可见光分光光度计测定RNA的浓度及A260/A280比值(表4)，A260/A280比值均在1.8-2.2之间，表明提取得到斑马鱼总RNA质量较好，可用于后续q-PCR实验。引物序列见表5。
表4.总RNA的浓度及A260/A280比值(n＝30)
Figure PCTCN2022116848-appb-000004
Figure PCTCN2022116848-appb-000005
表5.引物序列信息
Figure PCTCN2022116848-appb-000006
3.2col1a1a基因相对表达量和col1a1b基因相对表达量
详细见表6。
表6.各组col1a1a/col1a1b基因相对表达量检测结果(n＝30)
Figure PCTCN2022116848-appb-000007
Figure PCTCN2022116848-appb-000008
与正常对照组比较，*p<0.05，***p<0.001
由上述表6可知，实施例1-6与单独的维生素C、胶原蛋白肽和血橙粉相比均具有更优的效果，且具有显著性差异。因此将引起显著性差异的组别同实施例1-6进行了比较，不同字母表示具有显著性差异(P＜0.05)。证明组方虽然只有较低浓度的胶原蛋白肽，但可以表现同高浓度胶原蛋白肽更显著的效果。
实验例3美白、补水、抗皱功效测试
本实验例考察了本发明的护肤组合物在美白和抗衰老方面的功效
1.实验方法
1.1志愿者入选标准：18-49岁女性，上述女性志愿者的皮肤情况均为干性皮肤、有明显的干纹和细纹，皮肤缺乏弹性，皮肤暗沉，且有明显的色素沉着。上述志愿者在第0天经
Figure PCTCN2022116848-appb-000009
检测，双颊水分≤65AU，前臂水分＜45AU，根据SKINAGING ATLAS VOLUMN2-ASIAN TYPE(by R.Bazin&F.Flament,ver.2010,MED’COM Press)中公开的图谱评估皮肤老化现象，结果为鱼尾纹和毛孔评级≥2级。
1.2.服用方法：
实施例7：每天服用3g；
实施例8：每天服用30mL；
上述每个实施例组的志愿者人数分别为30例。连续服用8周，在第0,4,8周时进行相关检测，进行前后变化分析。
1.3测试部位及测试项目：双颊水分、前臂水分、鱼尾纹部位皮肤弹性、全脸黑色素含量。
测试水分：采用
Figure PCTCN2022116848-appb-000010
(C+K,Germany)，测量双颊及前臂的水分，测量三次取平均值；
鱼尾纹部位皮肤弹性测试方法：采用MPA580(C+K,Germany)，测量鱼尾纹部位的皮肤弹性，测量三次取平均值；
全脸黑色素含量：采用MPA580(C+K,Germany)，测量双侧脸颊部位的黑色素含量，测量三次取平均值；
实验数据采用SPSS 21.0对第0周(基线)、第四周、第八周进行重复测量方差分析 检验和广义估计方程(GEE)回归分析。
2.实验结果
详细见表7。
表7.各组美白、补水、抗皱功效实验结果
Figure PCTCN2022116848-appb-000011
***与服用前比较，p<0.001
由上表7可知，相比服用前，服用本发明护肤组合物，可以显著提高双颊水分、前臂水分、鱼尾纹部位皮肤弹性，显著降低全脸黑色素含量，在美白、补水和抗皱功效方面获得了显著的效果。
显然，上述实施例仅仅是为清楚地说明所作的举例，而并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。

Claims (10)

  1. 一种护肤组合物，其特征在于，由如下重量份的组分组成：
    血橙粉1-50份，胶原蛋白肽10-1000份，和维生素C 0.1-50份。
  2. 根据权利要求1所述的护肤组合物，其特征在于，由如下重量份的组分组成：
    血橙粉1份，胶原蛋白肽1000份，和维生素C 0.1份；或
    血橙粉50份，胶原蛋白肽10份，和维生素C 50份；或
    血橙粉25份，胶原蛋白肽500份，和维生素C 50份。
  3. 一种护肤组合物，其特征在于，由如下重量份的组分组成：
    血橙粉1-50份，胶原蛋白肽10-1000份，维生素C 0.1-50份，透明质酸钠0.1-50份，和烟酰胺0.01-20份。
  4. 根据权利要求3所述的护肤组合物，其特征在于，由如下重量份的组分组成：
    血橙粉1份，胶原蛋白肽1000份，和维生素C 0.1份，透明质酸钠0.1份，烟酰胺10份；或
    血橙粉50份，胶原蛋白肽10份，和维生素C 50份，透明质酸钠50份，烟酰胺0.01份；或
    血橙粉25份，胶原蛋白肽500份，和维生素C 50份，透明质酸钠25份，烟酰胺20份。
  5. 根据权利要求1-4任一项所述的护肤组合物，其特征在于，所述的血橙粉为塔罗科、莫罗和/或桑吉耐洛的血橙的提取物。
  6. 根据权利要求5所述的护肤组合物，其特征在于，所述血橙粉的制备方法，包括：取血橙，低温冷榨，得到血橙汁；
    将所述血橙汁过滤，浓缩，添加载体，干燥；
    可选的，所述低温冷榨温度为0-5℃；
    可选的，所述浓缩步骤中采用低温真空干燥进行浓缩，温度35～45℃，真空度0.095～0.1MPa；
    可选的，所述载体包括：麦芽糊精、菊粉或抗性糊精；所述载体占血橙粉的质量百分比为40-60％；
    可选的，所述干燥为喷雾干燥。
  7. 根据权利要求1-6任一项所述的护肤组合物，其特征在于，所述胶原蛋白肽为由牛皮或猪皮中提取得到的；
    可选的，所述胶原蛋白肽的平均分子量小于2000Da。
  8. 一种如权利要求1-7任一项所述的护肤组合物的制备方法，其特征在于，按照配方称取各组分的原料，混合；或
    按照配方称取各组分的原料，加入水中，溶解，搅拌，灭菌；
    可选的，所述水的温度为70-90℃；
    可选的，所述搅拌条件为80-110r/min的转速，搅拌240～360秒，直至完全溶解，以40-50r/min的转速继续搅拌20-50分钟；
    可选的，所述灭菌条件为110～123℃条件下高温瞬时灭菌5～10s。
  9. 一种制剂，其特征在于，以权利要求1-7任一项所述的护肤组合物或权利要求8所述的方法制备得到的护肤组合物为活性成分；
    可选的，还包括制剂允许的赋形剂或载体；
    可选的，所述制剂的形式包括液体制剂和固体制剂；
    可选的，所述制剂的形式包括注射剂、片剂、胶囊剂、颗粒剂、粉剂或膏剂。
  10. 如权利要求1-7任一项所述的护肤组合物、权利要求8所述的方法制备的护肤组合物或权利要求9所述的制剂具有如下任意一项的用途：
    a.在制备具有护肤作用的药物或食品中的用途；或
    b.在制备具有美白皮肤作用的药物或食品中的用途；或
    c.在制备具有皮肤补水作用的药物或食品中的用途；或
    d.在制备具有延缓衰老作用的药物或食品中的用途；或
    e.在制备具有减少皮肤黑色素作用的药物或食品中的用途；或
    f.在制备具有减少皮肤细纹、紧致皮肤和/或提高皮肤弹性作用的药物或食品中的用途；或
    h.在制备抑制酪氨酸酶活性的药物或食品中的用途；或
    i.在制备调控col1a1a基因和/或col1a1b基因的表达水平的药物或食品中的用途。
PCT/CN2022/116848 2022-08-11 2022-09-02 一种护肤组合物和制备方法及其用途 WO2024031762A1 (zh)

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