WO2004078155A1 - Utilisation d'un extrait total de flavonoide d'une feuille de bambou dans des compositions cosmetiques comme facteur anti-vieillissement - Google Patents

Utilisation d'un extrait total de flavonoide d'une feuille de bambou dans des compositions cosmetiques comme facteur anti-vieillissement Download PDF

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WO2004078155A1
WO2004078155A1 PCT/CN2004/000136 CN2004000136W WO2004078155A1 WO 2004078155 A1 WO2004078155 A1 WO 2004078155A1 CN 2004000136 W CN2004000136 W CN 2004000136W WO 2004078155 A1 WO2004078155 A1 WO 2004078155A1
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skin
bamboo
bamboo leaf
flavonoids
cosmetics
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PCT/CN2004/000136
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Chinese (zh)
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Ying Zhang
Xiaoqin Wu
Boyi Lu
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Zhejiang University (Hangzhou) Leaf Bio-Technology Co., Ltd.
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Publication of WO2004078155A1 publication Critical patent/WO2004078155A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/005—Antimicrobial preparations
    • 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)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
    • A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K8/9771—Ginkgophyta, e.g. Ginkgoaceae [Ginkgo family]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9794—Liliopsida [monocotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/10—Washing or bathing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/74—Biological properties of particular ingredients
    • A61K2800/75—Anti-irritant

Definitions

  • the invention belongs to the field of daily cosmetics.
  • the invention relates to the application of bamboo leaf total flavones as anti-aging skin care factors in cosmetics.
  • bamboo leaf total flavones As a natural plant-derived anti-aging skin care factor, bamboo leaf total flavones have anti-free radical, antioxidant, anti-radiation, antibacterial, bacteriostatic and whitening, anti-wrinkle, and spotting effects. They can be used in daily cosmetics. application. Background technique
  • Cosmetics are very popular commodities, and their consumption follows the trend of the society and has distinctive characteristics of the times. With the deepening of medical research, consumers are more cautious about using cosmetics made of chemical compounds. In recent years, cosmetics have begun to naturalize raw materials, forming a world boom in the development of natural resources. With the improvement of the quality of life that comes with economic development, how to delay the aging process has become the hottest topic.
  • Skin is an important part of maintaining human life and health. Direct contact with the outside world is the first barrier for the body, and it is extremely susceptible to damage caused by environmental factors. In addition to aging factors caused by age, stress, pollution, sunlight and ultraviolet radiation are all important causes of wrinkles or wrinkles on the skin.
  • Skin aging is mainly manifested in two aspects: (1) the skin gradually loses its elasticity and forms wrinkles; (2) the appearance of the skin gradually produces various pigmentations such as melasma and age spots.
  • free radicals free radical damage to the skin is an important factor in the process of skin aging.
  • Excessive reactive oxygen free radicals in the body interact with unsaturated fatty acids to generate substances such as malondialdehyde (MDA).
  • MDA malondialdehyde
  • Free radicals can also cause the collagen fibers and elastic fibers in the skin to cross-link and denature, become brittle, and lose elasticity.
  • the elastic fibers When there is insufficient moisture in the skin, it is easy for the elastic fibers to break and form wrinkles.
  • the deeper causes of skin aging also include slower renewal metabolism of epidermal keratinocytes and reduced proliferation and division of dermal fibroblasts. Compared with young people, the skin of the elderly is thinner, and the skin keratinocytes and fibroblasts are significantly reduced. Therefore, promoting the proliferation of aging skin cells is one of the goals of many skin care products. If the chemical can reduce the generation of skin free radicals, increase the activity of SOD, and have a certain proliferation effect on skin cells, it can accelerate the regeneration of skin tissue, increase the production of collagen fibers, protect the skin's moisture, and effectively slow down the skin. Aging [Huang Hansheng, Anti-aging Skin Care Products, Daily Chemical Industry Translation Series, 1994 (2): 35-38].
  • Cosmeceuticals meaning functional nutritional cosmetics.
  • the age characteristics of the aging population and the special industry background in which cosmetics are located provide a broad space for the application of plant extracts with physiological activities such as anti-free radical, anti-oxidation, anti-radiation, antibacterial, and anti-inflammatory. Coupled with the chain reaction brought by Mad Cow Disease in recent years, animal-derived skin care factors can easily cause people's psychological panic, so the demand for phytochemicals is increasing. There are more than 50 types of plant-based makeup and beauty products launched at home and abroad according to the types of plants added. The specific effects can be summarized as anti-wrinkle, whitening, freckle, itching, moisturizing, weight loss and so on.
  • the extracts of these natural plants have the characteristics of small side effects and good curative effects, and the cosmetic products made have not only maintained the characteristics of the cosmetics, but also have good therapeutic and health effects.
  • Flavonoids are widely distributed in the plant kingdom. So far, more than 2,000 species have been discovered, most of which exist in the form of glycosides and a few exist in the free form. Their physiological effects and biological activities are diverse. Diverse. Due to its conjugate structure, it shows strong absorption to both ultraviolet and visible light, and is highly stable in the visible and ultraviolet regions. Naturally derived flavonoids also have antibacterial, anti-photosensitivity, anti-oxidation, detoxification, and whitening effects. They can remove free radicals in the skin, avoid damage to cells by free radicals, promote skin metabolism, improve skin elasticity, and delay skin.
  • phospholipids can treat dermatitis; strong penetration on the skin, can inhibit hyaluronidase activity, and the use of hyaluronic acid can prolong the physiological activity of the substance; can inhibit and eliminate skin pigmentation (such as age spots).
  • a small amount of luteolin (0.01%) in the nail polish can keep the nail glaze surface smooth.
  • Rutin has anti-oxidant, anti-inflammatory and anti-stomatitis virus effects.
  • Rutin and limonene in a 50: 1 combination have a good inhibitory effect on active oxygen, can strongly absorb ultraviolet rays, can be used for sunscreen, whitening, hair care cosmetics [Wang Jianxin, Natural Active Cosmetics, China Light Industry Press, 1997: 214 ].
  • Adding 0.1 to 0.2% soy isoflavones to whitening cosmetics can prevent skin aging, blackening and brown spots; adding 2% to hair cream can absorb ultraviolet rays and maintain hair gloss.
  • Delphinium in anthocyanins has strong anti-oxidant and anti-inflammatory effects, and can inhibit bacterial infections caused by skin deficiency caused by excessive sebum secretion in scalp or skin.
  • the addition of cyanogenin in cosmetic water can provide better consistency and smoothness, and has deodorizing and astringent effects.
  • Flavonoids because of It has a variety of activities and its non-toxic and harmless safety and has a wide application prospect in medicines, foods and cosmetics [Huang Xingyu, Ding Jianquan, the role and mechanism of quercetin and its derivatives in cosmetics , Spices, Fragrances, Cosmetics, 1998 (3): 17-18].
  • Ginkgo flavonoids are powerful active oxygen free radical scavengers, which can protect skin cells from the effects of peroxidation, thereby prolonging the life span of skin cells and enhancing their anti-aging ability. Ginkgolides can also accelerate metabolism, improve blood circulation, and enhance Cell viability; In addition, Ginkgo biloba extract also has a broad-spectrum bactericidal effect, and has a significant inhibitory effect on common skin-infecting pathogens such as Staphylococcus aureus, Pseudomonas aeruginosa, and Floccus epidermatis The effect can be displayed at very low concentrations. This highly effective antibacterial characteristic is extremely advantageous for its use as a functional cosmetic ingredient.
  • tea extracts have been added to toothpastes and shampoos for more than a decade.
  • Mitsui Agriculture and Forestry Corporation of Japan has launched a variety of products containing tea polyphenols (tea catechins) for food and cosmetics.
  • Flavonoids in tea are mainly flavanols and flavonols, and their main functions are anti-oxidant, anti-cancer, anti-microbial, and deodorant.
  • Catechin belongs to baicalol, which accounts for 20-30% of the dry weight of green tea.
  • the main flavonols in tea include quercetin, tetrahydroxyflavonoids, and myricetin flavonoids, which account for about 2 ⁇ 3% of the water-soluble extract of tea.
  • catechins delay tooth decay (anti-caries) and improve breath freshness.Therefore, it can be added to toothpaste, mouthwash, chewing gum and breath freshener, and can also be applied to other daily necessities In many shampoos, moisturizing creams, perfumes, and sunscreens, tea extracts are contained. It is believed that they can calm the skin and act as an antioxidant to protect the skin from free radical attacks. [Jia Xudong, Tea Flavonoid Function And its application, Foreign Medical Hygiene Section, 2001, 28 (6): 369-371].
  • the main components of puerariae include isoflavones, triterpenoid saponins, and alkaloids, among which the most concerned are isoflavones such as puerarin and daidzein.
  • Isoflavones in Kudzu root can significantly inhibit the catalytic activity of tyrosinase, interrupt the melanin oxidation process, inhibit the occurrence and formation of melanin, and thereby prevent pigmentation such as melasma and sun spots. Therefore, Pueraria lobata is praised by the international cosmetics industry as another skin decolorizing component derived from green plants, and is used in freckle cosmetics by Japan, which is leading in cosmetic technology. [Tang Chunhong, Chen Qi, Current status of research and development of nutritional health functions of Kudzu at home and abroad, China Food Additives, 2002, (6): 56 ⁇ 58].
  • Licorice flavonoids in an oil-soluble extract from licorice root have an inhibitory activity against acne on the skin.
  • the development of its water-soluble preparation has also been successful, and the prevention and treatment of acne and acne have been proven on the human body: for testosterone-5a reductase-induced acne and androgen-induced acne (anti-androgen receptor) Both have a preventive effect; it also has a preventive effect on acne caused by acne bacteria, and also has an effect on acne caused by lipase and phospholipase [Yamamoto Jin, information on new raw materials for cosmetics-water-soluble preparation of licorice flavonoids, 2001, ⁇ 12-13]. ⁇
  • flavonoids such as hesperidin
  • hesperidin can prevent erythema and skin cancer caused by UV-induced skin cell lipid peroxidation, which means that it can be used as an ideal raw material for sunscreen cosmetics.
  • the effective ingredients of grass coral contain flavonoid glycosides, coumarins, organic acids, etc.
  • Jiangxi grass coral group company has tested its extracts in products such as shampoo, facial cleanser, shower gel, sun cream and toothpaste [ ⁇ , ⁇ Coral flow extract and its application in cosmetics, Daily Chemical Industry, 2000 (4): 61-64].
  • the purpose of the present invention is to provide a bamboo-derived anti-aging skin-care factor: bamboo leaf total flavonoids.
  • bamboo leaf total flavonoids In a first aspect of the present invention, there is provided an application of bamboo leaf total flavones in cosmetics, wherein the bamboo leaf total flavones are applied in cosmetics as an anti-aging skin care factor.
  • a cosmetic product which contains 0.001 to 10 wt% (more preferably 0.005 to 1 wt% of bamboo leaf total flavones.
  • the cosmetics are nutritional beauty lotions (creams), anti-wrinkle whitening honey, sunscreens, facial cleansers, shower gels, and shampoos.
  • Figure 1 is the infrared spectrum of bamboo leaf total flavonoids (compressed by potassium bromide);
  • Figure 2 is the UV spectrum of bamboo leaf total flavonoids (dissolved in spectrally pure methanol). detailed description
  • bamboo plants not only have high economic value, but also have extensive ecological and social benefits. With its unique biology, ecology, and multi-purpose characteristics, bamboo is increasingly being valued by people, and it is playing an increasingly important role in China's sustainable development strategy. .
  • bamboo leaf extract is Zhang It is a botanical flavonoid preparation developed in the 1990s. Its invention patent "a health beer with bamboo leaf flavonoid extract (ZL 98 1 04563.4)” and “extract flavonoid compound extract from bamboo leaves or The powder production method (ZL 98 1 04564.2) was authorized by the China National Patent Office in 2000 and 2001, respectively. A large number of studies have shown that bamboo leaf flavonoids have excellent biological effects such as anti-free radical, anti-oxidation, anti-aging, antibacterial, antiviral, protection of cardiovascular and cerebrovascular, prevention and treatment of senile degenerative diseases.
  • the functional factors of bamboo leaf extract are mainly C-glycoside flavonoids.
  • the four main bamboo leaf glycoside flavones are Orientin, Homoorientin, Vitexin, and Isolithin Glycoside (Is 0V it eX in).
  • glycoside flavones Compared with oxoflavones, glycoside flavones have the following outstanding advantages: (1) stable structure, not easy to be degraded; (2) can penetrate deep into the lesion site, and directly exert therapeutic effects; (3) enhanced hydrophilicity, which is beneficial to Development of food, medicine and cosmetics.
  • the international academic community has been paying attention to flavonoids since the 1990s, and this field is the latest research front.
  • mice can significantly enhance the resistance of mice to non-specific stimulation (normal pressure hypoxia test, p ⁇ 0.01) and anti-fatigue ability (swim test, p ⁇ 0.01); learning to normal mice Ability to promote to a certain extent (electric labyrinth method, p ⁇ 0.05) ; significantly induce the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) in aged mice Effect; significantly inhibits the generation of plasma peroxidized lipids (LPO) in elderly mice, and reduces the content of lipofuscin (LF) in the liver; by measuring changes in hydroxyproline content in mouse skin and tail tendon, low Positive effects on collagen were shown at the dose, indicating anti-oxidation, inhibition of lipid peroxidation products, elimination of lipofuscin, and protection of collagen [ZHANG Ying, TANG Lili, Anti-aging effect of Maojin bamboo leaf extract Experimental Research, Bamboo Research Transactions,
  • the present invention also by means of experimental weak chemiluminescence and fluorescence spectrophotometry, etc., a comparative study of tea polyphenols, bamboo leaf extract of Ginkgo biloba, and three plant Total Flavonoids Flavonoids Clear 02-- ⁇ Its activity and 6Q Co -y irradiation and CuS0 4 -Phen-Vc- H 2 0 2 -DNA system protects DNA damage caused by plutonium.
  • Tea polyphenols content 99%, batch number 991227, provided by Professor Yang Xianqiang, College of Agriculture and Biotechnology, Zhejiang University; Ginkgo biloba extract: total flavonoid glycoside content ⁇ 24%, batch number 20000041, produced by Ningbo Chinese Medicine Pharmaceutical Factory; total bamboo leaf flavonoids: total Flavonoid glycoside content ⁇ 24%, batch number 980625, provided by Professor Zhang Ying, School of Biological System Engineering and Food Science, Zhejiang University.
  • the above samples were respectively prepared into a stock solution of 1 mg / mL with three distilled water, and were set aside.
  • Calf thymus DNA, hypoxanthine, xanthine oxidase, luminol, phenanthroline (CPhen), and fluorescent probe (EB) are products of Sigma, USA; potassium nitrate, copper sulfate, PBS and CBS Drugs and reagents are analytically pure; yeast polysaccharide suspensions are made from active dry yeast; Vc and hydrogen peroxide solutions are now in use.
  • RF-5000 fluorescence spectrophotometer produced by Shimadzu Corporation, Japan
  • BPCL-II ultra-weak luminometer manufactured by the Institute of Biophysics, Chinese Academy of Sciences
  • ⁇ ⁇ irradiation source was provided by the irradiation center of the Institute of Nuclear Agriculture, Zhejiang University.
  • Vc-Cu 2+ -H 2 O 2 -yeast polysaccharide system was used to generate rhenium, and the hypoxanthine-xanthine oxidase-lumino system was used to generate 0 2 _. It was inhibited by samples with appropriate concentration range, ultra-weak light emission. Measure the luminescence value of the blank and test system, calculate the inhibition rate, and further calculate the half-values of ⁇ and 0 2 _ for different samples by linear regression equation. Inhibition concentration (IC 5. ). '
  • Flavonoids may compete with EB for binding sites on DNA molecules and pull down EB molecules embedded in DNA base pairs. In this way, the higher the concentration of the sample, the fewer EB molecules bound to the DNA, and the EB -The lower the fluorescence intensity of DNA.
  • Table 3 shows that irradiation can have direct and indirect effects on the DNA solution.
  • Direct effects can cause DNA molecule hydrogen bond destruction, double helix unraveling, base shedding, single and double-strand breaks, etc.
  • Indirect effects are mainly due to free radicals generated during irradiation, especially tritium, which attack the DNA strands and cause DNA molecules Damage. As a result, the binding sites of EB and DNA are reduced, and the fluorescence intensity value is also reduced.
  • Table 4 show the effects of different kinds and different concentrations of samples on the EB-DNA fluorescence intensity at different doses. Flavonoids can effectively scavenge free radicals, and therefore have a significant protective effect on the free radical damage of DNA molecules caused by irradiation. Tea polyphenols, ginkgo biloba extract, and bamboo leaf total flavones can maintain the fluorescence intensity of EB-DNA at a high level at different concentrations in the range of 20 ⁇ 160 ⁇ ⁇ / ⁇ , indicating that they have protection DNA irradiation damage effects and show concentration dependence. From the perspective of the protective effects of the three extracts, tea polyphenols were slightly stronger than the ginkgo extracts and bamboo leaf total flavones, and the latter two were close.
  • the chemiluminescence system of CuSO 4 -Phen-Vc-H 2 O 2 -DNA is in an alkaline medium, and phenanthroline (Phen) can react with H 2 O 2 to generate chemiluminescence under the catalysis of metal ions.
  • the emission wavelength is in the range of 445 ⁇ 450nm. According to relevant literature reports, 0 2 ⁇ is the main factor that causes Phen to generate chemiluminescent substances, and it is in the front peak position on the luminescence kinetic curve.
  • the plutonium generated in the system causes DNA damage and breakage, and produces a slow chemiluminescence that is delayed from the light emission of Phen itself.
  • the maximum emission wavelength is between 380 ⁇ 420nm. This luminescence is a characteristic reaction of free guanine. The top is in the rear peak position.
  • the peak value of the front peak and the peak of the rear peak decreased significantly, and its semi-inhibitory concentration was about 0.2 g / mL.
  • the decrease in the luminescence peak is due to the elimination of ⁇ and the like that initiated the chain reaction by the antioxidant, which reduces the DNA damage caused by ⁇ and the like. It is believed that the flavonoid acts as a preventive antioxidant.
  • the appearance time of the front peak value has almost no change, and the latter peak value lags behind, and with the increase of the antioxidant concentration, the peak backward time is prolonged.
  • ICR mice (clean grade), weighing 18 ⁇ 21g, males, were provided by Zhejiang Experimental Animal Center.
  • the total flavonoids in bamboo leaves is ⁇ 50%. It is a freeze-dried tan-like crystalline powder, stored in a desiccator, and prepared with distilled water before use.
  • 2% croton oil Formulated with 2% croton oil, 20% absolute ethanol, 5% distilled water, and 73% diethyl ether when in use; Indomethacin: API, gifted by Ningbo Pharmaceutical Factory, 1% CMC when used -Na suspension formulation; Xylene: Hangzhou Chemical Reagent Factory; MH broth and MH agar were purchased from the Logistics Inspection Factory of Zhejiang Military Region; 0.22 ⁇ m bacterial filter membrane was produced by the American company Milipore; newborn calf serum: super No mycoplasma, purchased from Hangzhou Sijiqing Bioengineering Materials Research Institute.
  • Shell / JB 3200 CO 2 culture phase product of the American Sheldon company; DG-3022A enzyme-linked immunoassay, produced by East China Electron Tube Factory; COIC biological inverted microscope, manufactured by Chongqing Optical Instrument Factory; SC-1 horizontal laminar flow purification Workbench, manufactured by Suzhou Institute of Clean Technology; CQ250 ultrasonic cleaner, produced by the 726 Ultrasonic Instrument Factory of the Seventh Hospital of China Shipbuilding Corporation.
  • Streptococcus pyogenes is performed according to the drug liquid dilution method: 10% of inactivated calf serum is added to the sterilized MH broth, and then the sample is added to make a concentration of 25mg / mL, and the bacteria filter is installed in a sterilized large syringe. Sterile filtration treatment. The sterile filtrate was diluted twice with sterile MH serum medium, 1 mL per tube. Add 0.05mL of young Streptococcus pyogenes cultured for 4 to 6 hours (total bacteria content is 10 4 cfu) and incubate in a 35 ° C incubator for 20 to 24 hours. Tube as its MIC value.
  • bacteria are carried out by double dilution of the drug: after dissolving each vial of MH agar, autoclaving and dissolving, cooling to about 55 ° C, adding different amounts of samples to make them 25.0, 12.5, 6.25, 3.125, respectively. , 1.56, 0.78 and 0.39 mg / mL concentration, set a MH agar without drug as a positive growth control dish after adding bacteria. Then add 6 kinds (see 3.1.1 strain source) bacterial solution of turbidimetric concentration to each dish (total bacterial content is 10 4 cfu), and use the inoculation ring to paint small garden spots (both are 4 ⁇ 6h juvenile bacteria), cover the dish after drying, and incubate at 35 ° C for 16 ⁇ 20h. Use the lowest sample concentration tube with no bacterial growth as its MIC value. Table 7 Antibacterial effect of total flavonoids in bamboo leaves
  • mice were randomly divided into 5 groups according to body weight, 12 in each group. The settings were as follows: 1 Control group: equal volume of normal saline, 0.8mL / 20g; 2 Indomethacin group: indomethacin 10mg / kg; 3 high-dose group: 10mg / mL BLTF-01 at a dose of 400 mg / kg; 4 Medium dose group: 5 mg / mL BLTF-01 at a dose of 200 mg / kg; 5 Low dose group: 2.5 mg / mL BLTF-01 at a dose of 100 mg / kg.
  • mice were po-administered once a day for 7 days (indomethacin was administered only once), and lh after the last administration, mice were coated with 2% croton oil 0.02mL / head in the right ear, causing inflammation, and sacrificed after 4h, along the auricle Cut the left and right ears, punch the left and right ear pieces with a 9mm ring drill, weigh them, and record the difference between the weights of the left and right ear pieces as the swelling rate.
  • mice were given 200 mg / kg and 400 mg / kg of bamboo leaf total flavonoids by gavage for 7 consecutive days, and had significant (* p ⁇ 0.05) and extremely significant (p ⁇ 0.01) on croton oil-induced auricle swelling.
  • the inhibitory effect was significant and showed a significant dose-dependent relationship (Table 8).
  • mice were randomly divided into 5 groups according to body weight, with 10 mice in each group. The setting was the same as 3.2.2.1. Animals were administered once a day for 7 consecutive days (indomethacin was administered only once), and 30 minutes after the last administration, the mice's right ear was coated with xylene 0.05 mL / head, causing inflammation, After 15 minutes, the rats were sacrificed, the left and right ears were cut along the auricle, and the left and right ear pieces were punched out with a 9mm ring drill, weighed, and the difference between the weights of the left and right ear pieces was recorded as the swelling rate. Compare with the control group to judge the curative effect.
  • mice were given a total of 400 mg / kg of bamboo leaf total flavones by gavage for 7 consecutive days, which had a significant (* p ⁇ 0.05) inhibitory effect on auricle swelling caused by xylene. (Table 9).
  • bamboo leaf total flavonoids brownish yellow powder with a total flavonoid content of 28.7%.
  • the sample was dissolved in DMSO, filtered through a 0.22 ⁇ m filter to remove bacteria, and diluted with serum-free DMEM to the test concentration. The final concentrations were 0.5%, 0.05%, and 0.005%, and stored at -4 ° C. Serum-free DMEM was used as a control.
  • DMEM medium keratinocyte (K-SFM) medium (Gibco BRL), insulin (American Sigma), trypsin (American Difco) calf serum (NBS, Hangzhou Sijiqing Company), tetramethyl coupling Azole (MTT, Fluka Corporation, USA), dimethyl sulfoxide (DMSO, Shanghai Institute of Biochemistry); cortisone, penicillin, streptomycin, Ficoll, 96-well plate and 24-well plate, and 35mm culture dish and 25cm2 culture flask (Coming, USA).
  • Microplate reader BIO-TEK
  • pH meter clean bench, incubator, 722 spectrophotometer.
  • the number of cells was adjusted to 1 ⁇ 10 4 cells / ml with DMEM medium containing 15% NBS, and inoculated into culture flasks and 35 mm petri dishes.
  • the 96-well plate was seeded with 2 ⁇ 10 5 cells per well. After the cells were cultured in a sub-fusion state, the solution was changed and test substances of various concentrations were added. Each concentration was 4 samples. After adding the drug, 10 ⁇ MTT was added to each well for 24 hours, the culture was continued for 4 hours, and the supernatant was discarded. After adding 100 ⁇ of the lysate, it was detected on a microplate reader at 570 mm and 630 nm.
  • MDA was determined by TBA colorimetry, SOD was determined by nitrite reduction, and the kit was provided by Nanjing Jiancheng Biological Company.
  • the density of B 16 cells was adjusted to 1.0 ⁇ 10 4 cells.
  • Each 35 mm culture dish was inoculated with 1.0 mL of a cell suspension and cultured for 24 h.
  • test substances of various concentrations were added, the cells were harvested at 24 h, washed twice with PBS, 2.0 M1 PBS (containing 0.1% Triton-X) was added, and the cell debris solution was removed at 37 ° C for 30 min. 1mL for protein determination.
  • Add 2.0 mL of PBS containing 0.1% L-DOPA to the remaining solution, incubate at 37 ° C for 1 h, and determine the colorimetry at 400 nm.
  • the cell density was adjusted to 1.0 x 10 4 cells, and the cells were seeded in a culture flask for 24 hours, the test substance was added, the L solution was changed at 48 h, and the cells were harvested at 72 h. The cells were washed with PBS and trypsinized. The collected cells were lysed in 2 mL of PBS, and 0.2 mL was taken out to determine the protein content. To the remaining 0.9 mL of the cell fluid, 1.8 mL of 4 mol / L NaOH was added, and the mixture was heated at 100 ° C for 20 minutes, and the colorimetric measurement was performed at 400 nm.
  • test substance concentration is 10%.
  • Skin irritation test Take 0.2ml of test substance and apply it on the skin. Once a day for 1 h each time for 14 consecutive days.
  • Eye irritation test 0.1 ml of the test substance was dropped into the conjunctival sac, and the eyes of the animals were tested at 1, 24, 48, 72 h, and on the 4th and 7th days after exposure. an examination.
  • Table 10 shows that the total flavonoids of bamboo leaves promote skin cell proliferation in a dosage range of 0.005% to 0.05%, wherein the proliferation effect on skin keratinocytes is significant at a concentration of 0.005% (p ⁇ 0.05) and at a concentration of 0.05% Very significant (p ⁇ 0.01); there was also a significant difference in the proliferation of skin fibroblasts compared with the control group (at 0.005%, p ⁇ 0.05).
  • B16 melanoma tumor cell tyrosinase inhibition test results show that the total flavonoids of bamboo leaves have no significant effect on the enzyme activity; and from the results of melanin synthesis tests, it has been shown that the concentration of melanin is significant at 0.005% ⁇ 0.05% Inhibition (see Table 12).
  • the skin irritation test and eye irritation test of bamboo leaf total flavones were negative, showing no skin and eye irritation.
  • the skin irritation test and eye irritation test are to understand whether foreign substances have irritating and damaging effects on the skin or cause inflammatory lesions of the mucosa in the case of contact with the skin or mucous membranes. They are mandatory inspection items for the safety evaluation of cosmetics and their raw materials.
  • the test results showed that the skin and eye conjunctival irritation response of total bamboo leaf flavonoids were negative, indicating that under external skin application conditions, it would not cause irritation to human skin and mucous membranes.
  • Tests show that bamboo leaves total flavonoids have good biological effects on skin cells, can promote the proliferation of skin keratinocytes and fibroblasts, increase SOD activity, and reduce oxidative damage, suggesting that bamboo leaves total flavones can be used to delay skin aging.
  • the active substance is applied to cosmetics.
  • the skin whitening effect test found that bamboo leaves total flavonoids can reduce the amount of melanin synthesized by melanocytes, suggesting that it may have a whitening effect on the skin.
  • the total flavones of bamboo leaves referred to in the present invention are flavonoid preparations with different accuracy obtained from the leaves of Graminae, Bambusoideae: and Phyllostochys Sieb. Et Zucc varieties, and the production process thereof It has been involved in two previous invention patents of Zhang Ying (patent numbers ZL 98 1 04564.2 and ZL 98 1 04563.4). It should be noted that the bamboo leaf total flavonoids referred to in this patent can be either products obtained by using the above patent process, or based on this, further use of high-tech technologies such as adsorption-desorption, membrane separation, supercritical fluid extraction and the like The obtained bamboo leaf flavonoid product is refined by a combined method.
  • total flavonoids in bamboo leaves is yellow or brownish-yellow powder (also available in the form of extract), and the total flavonoid glycoside content (on a dry basis) can vary between 10 and 90%, among which humengin, isohumuloside
  • the content of the four main glucosides, vitexin, and isovitein is more than 50% of the total flavonoid glycosides.
  • the infrared spectrum of the potassium bromide tablet shows that bamboo leaves total flavonoids have characteristic absorption near 3400, 2900, 1610, 1520, 1080cm- 1 and so on (see attached picture 1); after dissolving them in pure methanol Scanning in the wavelength range of 200 ⁇ 600mn, the ultraviolet spectrum shows that there are two main absorption peaks in the 240 ⁇ 400nm region, of which there is a strong absorption peak between 240 ⁇ 280nm, and once between 300 ⁇ 350nm Strong absorption peak, which meets the typical characteristics of flavonoids (see Figure 2).
  • Identification of chemical reagents for total flavonoids in bamboo leaves Take 0.5g of the sample and dissolve it in 100mL of 95% ethanol. 1 Take 1mL of the above solution, and force 2 ⁇ 3 drops of 1% FeCl 3 -ethanol solution. It should be dark blue or blue-violet. 2 Take 1mL of the above solution, add 2 ⁇ 3 drops of 1% A1C1 3 -ethanol solution, it should be bright yellow. + Take 0.5g of BLTF, add 10mL of ether, ultrasonic-assisted extraction for 30s, and filter.
  • anti-free radical, antioxidant, anti-aging, anti-radiation, antibacterial, antibacterial, anti-inflammatory, whitening, anti-wrinkle functions of bamboo flavonoids it also meets the safety requirements as a cosmetic additive, and can be used as The functional factors of anti-aging skin care cosmetics are applied in various daily cosmetics such as nutrient moisturizers, sunscreen skin care agents, anti-wrinkle whitening agents, cleansing milks, shampoos, shower gels and so on.
  • the skin-care factor of the present invention can be appropriately blended with components commonly used in cosmetics to make products for various uses. These other components can include various vitamins and their derivatives, hyaluronic acid, surfactants, wetting agents, chelating agents, pH adjusters, excipients, other UV absorbers, preservatives, dyes, fragrances, etc. .
  • the cosmetics of the present invention can be made into various forms, such as oily cosmetics, emulsion cosmetics, and water-based cosmetics.
  • the cosmetics of the present invention have cosmetic skin care effects such as whitening, anti-wrinkle, sun protection, anti-inflammatory, mottled spots and cowpea.
  • the main advantages of the present invention are: It provides a broad-sourced, safe, efficient, and economically applicable anti-aging skin-care factor—total flavonoids from bamboo leaves, and systematically studies its characteristics related to the physiological functions of the skin, showing that it has anti-freedom Base, anti-oxidation, anti-radiation, antibacterial, bacteriostatic, promote skin cell proliferation, prevent skin oxidative damage, reduce melanin synthesis, delay skin aging and other beauty effects, and can give the product a unique fragrance of bamboo, such as nourishing and nourishing Agents, sunscreen skincare agents, anti-wrinkle whitening agents, cleansing milk, shampoo, shower gel and other daily cosmetics fields have broad application prospects.
  • This product has antibacterial, cosmetic, improving blood circulation, promoting metabolism, enhancing skin vitality, moisturizing the epidermis, inhibiting the secretion of abnormal sebum, and activating cells and anti-tissue toxicity.
  • This product has anti-wrinkle, whitening, moisturizing, and spotting effects.
  • This product has sun protection, anti-aging and skin care effects.
  • Vitamin E and its derivatives 0.05 ⁇ 0.5
  • This product has bacteriostatic, inhibited secretion of sebum secretion, anti-inflammatory, cowpea and so on.
  • This product can inhibit harmful bacteria on the skin surface, activate skin cells, condition the skin and prevent skin cancer, and make the body with a pleasant fragrance for a long time after bath.
  • this product can also prevent the loss of keratin during shampooing, prevent the yellowing and bifurcation of the hair, prevent itching of the scalp, and protect the hair. Hair has a light fragrance for a long time.

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Abstract

Cette invention porte sur l'utilisation de l'extrait total d'un flavonoïde d'une feuille de bambou dans des compositions cosmétiques comme facteur anti-vieillissement. L'objet de cette invention est de produire un facteur naturel anti-vieillissement. Le flavonoïde extrait des feuilles de bambou est efficace comme anti-radicaux libres, antioxydant, anti-radiation, antimicrobien, inhibiteur de bactéries, facilite la prolifération des cellules de la peau, prévient les lésions oxydatives de la peau, réduit la formation de mélanine, ralentit le processus de vieillissement de la peau, sans créer d'irritations et de réactions allergiques sur la peau et la muqueuse. Cette invention permet d'obtenir des compositions cosmétiques au parfum unique et frais de bambou et peut être utilisée dans différentes variétés de compositions cosmétiques telles que des produits nourrissants pour la peau, des écrans solaires, des antirides et des produits blanchissant la peau, des produits démaquillants et des lotions pour le bain, etc.
PCT/CN2004/000136 2003-03-03 2004-02-23 Utilisation d'un extrait total de flavonoide d'une feuille de bambou dans des compositions cosmetiques comme facteur anti-vieillissement WO2004078155A1 (fr)

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WO2013086649A2 (fr) 2011-12-13 2013-06-20 Universidad De Chile Flavonols utilisés comme agonistes de la coenzyme q (ubiquinone et ubiquinol) pour moduler l'activité des complexes de la chaîne de transport d'électrons au niveau mitochondrial
WO2018236304A3 (fr) * 2017-06-09 2019-03-28 Actv Biyoteknoloji Laboratuvar Sanayi Ve Ticaret Anonim Sirketi Matière première à base de plante efficace contre les imperfections cutanées et procédé pour sa production

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CN102309436A (zh) * 2011-05-19 2012-01-11 大连九羊食品有限公司 一种含有羊乳脂的面霜及其制备方法
CN102885728A (zh) * 2011-07-21 2013-01-23 新乡博凯生物技术有限公司 一种以金银花总黄酮为主的天然护肤剂原料及其制备方法
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CN103385896B (zh) * 2013-07-23 2015-03-25 钟春燕 一种细菌纤维素复合黄酮类化合物的制备方法
CN103599009B (zh) * 2013-10-25 2016-06-08 青岛文创科技有限公司 一种控油洗面奶
CN104042506B (zh) * 2014-06-17 2017-01-11 广州环亚化妆品科技有限公司 一种拟缺香茶菜提取物及其制备方法和在化妆品中的应用
CN104523472B (zh) * 2014-12-03 2017-12-22 烟台新时代健康产业日化有限公司 一种竹叶黄酮复合脂质体、制备方法及其应用
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WO2013086649A2 (fr) 2011-12-13 2013-06-20 Universidad De Chile Flavonols utilisés comme agonistes de la coenzyme q (ubiquinone et ubiquinol) pour moduler l'activité des complexes de la chaîne de transport d'électrons au niveau mitochondrial
WO2018236304A3 (fr) * 2017-06-09 2019-03-28 Actv Biyoteknoloji Laboratuvar Sanayi Ve Ticaret Anonim Sirketi Matière première à base de plante efficace contre les imperfections cutanées et procédé pour sa production

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