CN110731918B - Application of nitraria tangutorum bobr extract and skin external preparation containing nitraria tangutorum bobr extract - Google Patents
Application of nitraria tangutorum bobr extract and skin external preparation containing nitraria tangutorum bobr extract Download PDFInfo
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- CN110731918B CN110731918B CN201810803356.7A CN201810803356A CN110731918B CN 110731918 B CN110731918 B CN 110731918B CN 201810803356 A CN201810803356 A CN 201810803356A CN 110731918 B CN110731918 B CN 110731918B
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- tangutorum bobr
- nitraria tangutorum
- extract
- nitraria
- skin
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Classifications
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- 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
- 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
- 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
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Mycology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Engineering & Computer Science (AREA)
- Cosmetics (AREA)
Abstract
The invention provides application of a nitraria tangutorum bobr extract and a skin external preparation containing the nitraria tangutorum bobr extract. The application specifically relates to the application of the nitraria tangutorum bobr extract in preparing one or more of an antioxidant, a melanin synthesis inhibitor, an elastic fiber hydrolysis inhibitor and an anti-skin aging protein expression up-regulating agent. The Nitraria tangutorum bobr extract, especially Nitraria tangutorum bobr seed extract, has the effects of scavenging free radical activity, inhibiting tyrosinase activity, inhibiting elastase activity, improving anti-aging protein expression in human fibroblasts, regulating and controlling synthesis of collagen in dermis, and delaying aging.
Description
Technical Field
The invention relates to application of a nitraria tangutorum bobr extract and a skin external preparation containing the nitraria tangutorum bobr extract.
Background
With the continuous development and the gradual maturity of the skin care product market, more and more brands emerge, and the requirement of consumers for the efficacy of the skin care product is expected to see the practical and effective skin care effect, so the research level of the efficacy mechanism of the active substances of the skin care product needs to be improved, the explanation of the efficacy mechanism becomes deeper and deeper, the simple description of the images cannot be realized, the advanced biotechnology needs to be fused, the discovery of the skin biology is deepened continuously with the continuous development of the cytobiology technical level, and more targets and regulation mechanisms related to the functional structure of the skin are discovered.
The skin is the largest organ of the human body, covers the whole body, and mainly plays a role in protecting the body, removing sweat, sensing cold and heat, pressure and the like, so that various tissues and organs in the body are prevented from being invaded by physical, mechanical, chemical and pathogenic microorganisms.
Skin aging is classified into endogenous aging and exogenous aging. Intrinsic aging is natural aging, an aging process caused by irresistible physiological factors with increasing age, belongs to gene-level aging, and is obviously characterized by the appearance of wrinkles and the laxity of the skin. Extrinsic aging is mainly aging caused by environmental factors such as ultraviolet radiation, high temperature, cold, environmental pollution, nutritional deficiency, diseases, bad life habits, etc., wherein ultraviolet radiation is the most dominant factor, and is therefore called photoaging, which is manifested by roughness and dryness of exposed portions of the skin, increased thickness of wrinkles, irregular pigmentation, vasodilation, epidermal keratosis, dermal elastic fibrosis, accumulation of degradation products, etc., and can theoretically be prevented or slowed. The theory of free radical aging states that excessive free radicals are considered to be an important cause of skin photoaging, and because the skin is exposed to external environments such as ultraviolet rays, active oxygen generated by the ultraviolet rays and active oxygen in other external environments generate various free radicals in the skin, so that the skin is damaged by oxidative stress, and finally, the skin is aged.
In general, dermal aging is manifested by a reduction in thickness, a decrease in collagen and elastin synthesis, an increase in breakdown, and an increase in lytic enzyme activity. The dermis thickness decreases by 20-80% during aging, with gradual tissue and function degradation. Collagen is a protein with the largest content in the human body, can endow the skin with toughness, elasticity, a waterproof function and the like, is an important raw material substance for maintaining the shape and the structure of the skin and tissues and organs and repairing various damaged tissues, and is also a main component of extracellular matrix. The reduction of collagen synthesis is an important feature of aging skin. The most predominant collagen in the dermis is type I collagen (COL I), and type V Collagen (COLV), one of the most predominant collagens, is a collagen that forms regulatory fibrils, localized with the papillary dermis, the matrix surrounding the basement membrane, and copolymerized with COL I.
In the dermis, many proteins are first expressed in a precursor form and then converted to mature proteins by proteolytic processing for normal function. BMP-1 (bone morphogenetic protein-1) is a major protease in the assembly of extracellular matrix molecules, and is involved in this process, and mainly functions to cleave peptide chains of some proteins in the extracellular matrix (ECM), and modify the precursors of many biomolecules into structural proteins and active enzymes with biological functions, including many matrix molecules such as type I, type III, type V, type VII procollagen, Lipoxygenase (LOX), small leucine-rich proteoglycan (SLRP), etc. Simply, BMP-1 functionalizes collagen, ensuring that collagen is modified in an ideal manner during biosynthesis, resulting in the correct protein conformation.
Collagen is the gold standard in various indexes for resisting skin aging, but the efficacy and mechanism of most products on the cosmetic market are only about how to increase the collagen content (such as promoting generation, supplying and protecting). Recent studies have shown that the three-dimensional conformation and structure of collagen also have a significant effect on skin gloss. For the skin, collagen is a light propagation channel, and collagen fiber-mediated light scattering plays an important role in light propagation penetration inside the dermis. If the skin has high collagen content and normal and orderly arrangement structure, light can be directly projected into the skin, and the appearance of the skin has transparent luster; if the collagen is less, the structure is abnormal, and the arrangement is disordered, the diffuse reflection of the light path (the scattering effect of the collagen beam) is formed, so that the appearance is dark, yellow and matt.
Photoaging caused by UV exposure causes increased activity of skin elastase. Elastase is distributed in various tissues and cells, and the increase of elastase activity can cause the hydrolysis of skin elastic fibers, thereby accelerating the decomposition of collagen and elastin in dermis, leading the skin to lose elasticity, and causing skin aging phenomena such as wrinkles and the like.
In addition, the degree of pigmentation increases as a result of the stimulation by UV exposure. Is caused by the synthesis of pigment by melanocytes on the skin and its transfer to surrounding keratinocytes. The pigmentation process is a complex process, mainly caused by biochemical reactions in melanocytes, i.e. tyrosine generates dopaquinone under the action of tyrosinase as a catalyst, and melanin is formed by enzymatic catalysis or non-enzymatic oxidation. The key point is the synthesis of melanin, which is controlled by tyrosinase.
Therefore, the research on how to improve the anti-aging capability of the skin has important significance on delaying skin aging.
Extracts from natural plant sources are increasingly used in the field of skin care due to their long history of use, excellent multiple efficacy.
White spurs (Nitraria tandorum Bobr.) are plants of the genus Nitraria of the family Zygophyllaceae, also known as Dizao, Pangbai, desert cherry, Tanggute white spurs, and are mainly distributed in northern Shaanxi, western inner Mongolia, Ningxia, Gansu Hexi, Qinghai, Xinjiang and northeast of Tibet, in desert and semi-desert areas of lake basin sand, river terrace, mountain front plain sand, and clay lands with accumulated sand. Due to the long-term adaptive evolution of the harsh habitat, the nitraria tangutorum bobr has the biological characteristics of drought resistance, high temperature resistance, saline and alkaline resistance, barren resistance, sand burying resistance, severe cold resistance and the like, and the strong stress resistance is less common in nature. The nitraria tangutorum bobr has the traditional recorded effects of strengthening the spleen and stomach, helping digestion, soothing the nerves, relieving exterior syndrome, expelling toxin, promoting lactation, promoting fattening of livestock and the like. Recent research shows that the Nitraria seed oil can resist oxidation, fatigue, radiation and liver injury; the total flavonoids of Nitraria sibirica pall have protective effect on vascular endothelial cell injury. The nitraria seed contains 10-12% of crude fat, wherein the content of unsaturated fatty acid is as high as 95%, the content of linoleic acid is the highest, and the relative content is generally 65-70%, which is one of the most remarkable characteristics that the nitraria seed oil is different from other seed oils; in addition, the Nitraria tangutorum bobr seeds also contain substances such as VE, flavone, phytosterol, allantoin and the like.
Disclosure of Invention
The invention provides application of nitraria tangutorum bobr extract and a skin external preparation containing the nitraria tangutorum bobr extract, aiming at solving the problem that active substances capable of improving the antioxidant and anti-aging capacity of skin are lacked in the prior art. The nitraria tangutorum bobr extract disclosed by the invention is clear in action mechanism, safe and reliable, and has the effects of resisting oxidation, whitening and resisting aging.
The inventor discovers through a large amount of researches that: 1) the Nitraria sibirica pall extract has the function of removing free radicals; 2) the Nitraria tangutorum bobr extract has tyrosinase activity inhibiting effect; 3) the Nitraria tangutorum bobr extract has the effects of inhibiting the activity of elastase, improving the expression of anti-aging protein in human fibroblasts, regulating and controlling the synthesis of collagen in dermis, and is suitable for cosmetics with the effect of delaying aging.
The invention mainly solves the technical problems through the following technical means:
the invention provides an application of nitraria tangutorum bobr extract in preparing one or more of an antioxidant, a melanin synthesis inhibitor, an elastic fiber hydrolysis inhibitor and an anti-skin aging protein expression up-regulator.
In the present invention, the antioxidant may be a substance that helps to trap and neutralize free radicals, such as a free radical scavenger, as is conventional in the art.
In the present invention, the melanin synthesis inhibitor may be a substance that inhibits melanin synthesis, such as a tyrosinase inhibitor, which is conventional in the art.
In the present invention, the elastic fiber hydrolysis inhibitor may be a substance that inhibits the hydrolysis of elastic fibers, which is conventional in the art, such as an elastase inhibitor. Generally, hydrolysis of elastic fibers in the skin accelerates the decomposition of collagen and elastin in the dermis, causing the skin to lose elasticity and to develop skin aging such as wrinkles.
In the present invention, the anti-skin aging protein may be a protein that is conventional in the art and is associated with skin aging, such as collagen and elastin.
In the present invention, the up-regulator of anti-skin aging protein expression may be a substance conventionally used in the art to affect the expression of a protein associated with skin aging, such as one or more of an up-regulator of BMP-1 (bone morphogenetic protein-1) expression, an up-regulator of collagen I expression, and an up-regulator of collagen V expression.
It is well known to those skilled in the art that BMP-1 can be processed to modify procollagen types I, III, V and VII to functionalize the collagen.
In the present invention, the concentration of the Nitraria tangutorum bobr extract in the antioxidant is preferably 0.025-0.075 mg/mL, and more preferably 0.05 mg/mL.
In the present invention, the concentration of the nitraria tangutorum bobr extract in the melanin synthesis inhibitor is preferably 0.25 to 0.75mg/mL, and more preferably 0.5 mg/mL.
In the present invention, the concentration of the Nitraria tangutorum bobr extract in the elastic fiber hydrolysis inhibitor is preferably 0.5 to 1.5mg/mL, more preferably 1 mg/mL.
In the present invention, the concentration of the Nitraria tangutorum bobr extract in the up-regulator of anti-skin aging protein expression is preferably 0.05-1 mg/mL, such as 0.05mg/mL or 1 mg/mL.
When the anti-skin aging protein is BMP-1, the concentration of the Nitraria tangutorum bobr extract in the up-regulator of the expression of the anti-skin aging protein is preferably 0.05 mg/mL.
When the anti-skin aging protein is collagen I or collagen V, the concentration of the Nitraria tangutorum bobr extract in the up-regulator of the expression of the anti-skin aging protein is preferably 1 mg/mL.
In the present invention, the antioxidant, melanin synthesis inhibitor, elastic fiber hydrolysis inhibitor and anti-skin aging protein expression up-regulator are suitably used in cosmetics or skin care products.
In the present invention, the Nitraria tangutorum Bobr extract can be prepared by the conventional method in the art, such as the method of mixing Nitraria tangutorum Bobr with supercritical CO2Mixing, and extracting to obtain extractive solution.
Wherein, the extraction part of the nitraria tangutorum bobr can be any part of the nitraria tangutorum bobr which can be used for being used as a medicine, and preferably is nitraria tangutorum bobr seeds. The Nitraria tangutorum Bobr seeds are generally mature seeds of Nitraria tangutorum Bobr.
When the extraction part of the nitraria tangutorum bobr is nitraria tangutorum bobr seeds, the nitraria tangutorum bobr extract is a nitraria tangutorum bobr seed extract.
Wherein, preferably, the nitraria tangutorum bobr is pretreated, such as crushed; more preferably, the crushed material is further sieved, and the mesh number of the sieved material can be 40 meshes.
Wherein, the pressure of the extraction can be the conventional pressure in the field, such as 30 to 40MPa, preferably 35 MPa.
Wherein, the temperature of the extraction can be the conventional temperature in the field, such as 40-50 ℃, preferably 45 ℃.
Wherein the supercritical CO2The flow rate of (B) may be a flow rate conventional in the art, for example, 20 to 30kg/h, preferably 25 kg/h.
Wherein, the extraction time can be the time conventional in the art, such as 2-3 h, preferably 2.5 h.
Wherein the extract can be purified according to the conventional operation in the field, for example, the extract is subjected to macroporous adsorption resin chromatography.
The macroporous adsorption resin can be AB-8.
The eluent for chromatography may be an aqueous ethanol solution, the concentration of the aqueous ethanol solution is preferably 65 to 75%, more preferably 70%, and the percentage refers to volume percentage.
The volume of the eluent for chromatography may be 4-6 times the column volume, for example 5 times the column volume.
The chromatography can be further subjected to impurity removal treatment, for example, elution with water is carried out to remove impurities, and the elution volume of the water can be 1 time of the column volume; preferably, after the water elution is performed to remove impurities, 25-35% ethanol water solution with 5 times of column volume can be performed to remove impurities, for example, 30% ethanol water solution is performed to remove impurities, and the percentage refers to volume percentage.
Wherein, preferably, the preparation method of the nitraria tangutorum bobr extract comprises the following steps: mixing Nitraria tangutorum bobr seeds with supercritical CO2Mixing, extracting, wherein the pressure of extraction is 30-40 MPa, the temperature of extraction is 40-50 ℃, and supercritical CO is used2The flow rate of the extraction solution is 20-30 kg/h, and the extraction time is 2-3 h to obtain an extraction solution; more preferably, the preparation method of the nitraria tangutorum bobr extract comprises the following steps: mixing Nitraria tangutorum bobr seeds with supercritical CO2Mixing, extracting at 35MPa and 45 deg.C, and supercritical CO2The flow rate of the extraction is 25kg/h, the extraction time is 2.5h, and the extraction is obtainedAnd (4) taking the liquid to obtain the finished product.
The invention also provides application of the nitraria tangutorum bobr extract as an antioxidant active ingredient, a whitening active ingredient and an anti-aging active ingredient in cosmetics or skin care products.
Wherein the Nitraria sibirica pall extract is as defined above.
The invention also provides a skin external preparation, which contains 0.001-10% of the nitraria tangutorum bobr extract, wherein the percentage refers to the weight percentage (g/100mL) in the skin external preparation.
Wherein the Nitraria sibirica pall extract is as defined above.
Wherein, the content of the nitraria tangutorum bobr extract is preferably 0.0025 to 0.15 percent, such as 0.005 percent, 0.05 percent or 0.1 percent, and the percentage refers to the weight percentage in the skin external preparation.
When the nitraria tangutorum bobr extract is used as an antioxidant in the skin external preparation, the content of the nitraria tangutorum bobr extract is preferably 0.0025 to 0.0075 percent, more preferably 0.005 percent, and the percent refers to the weight percent in the skin external preparation.
When the Nitraria tangutorum bobr extract is used as a melanin synthesis inhibitor in the skin external preparation, the content of the Nitraria tangutorum bobr extract is preferably 0.025 to 0.075%, more preferably 0.05%, and the percentage refers to the weight percentage in the skin external preparation.
When the Nitraria tangutorum bobr extract is used as a hydrolysis inhibitor of skin elastic fibers in the skin external preparation, the content of the Nitraria tangutorum bobr extract is preferably 0.05 to 0.15%, more preferably 0.1%, and the percentage refers to the weight percentage in the skin external preparation.
When the Nitraria tangutorum bobr extract is used as an up-regulator of the expression of anti-skin aging proteins in the external preparation for skin, the content of the Nitraria tangutorum bobr extract is preferably 0.005 to 0.1%, for example, 0.005% or 0.1%, by weight in the external preparation for skin.
When the anti-skin aging protein is BMP-1, the content of the Nitraria tangutorum bobr extract is preferably 0.005%, by weight, in the external preparation for skin.
When the anti-skin aging protein is collagen I or collagen V, the content of the nitraria tangutorum bobr extract is preferably 0.1%, and the percentage refers to the weight percentage in the skin external preparation.
In the present invention, the external preparation for skin may further comprise other active ingredients, preferably, one or more of a moisturizing active ingredient, an antioxidant active ingredient, an oil-controlling active ingredient, a whitening active ingredient, and an anti-aging active ingredient. The other active ingredients are present in amounts conventional in the art.
The external preparation for skin may further contain a vehicle or a base excipient conventionally used in the art according to various formulations and purposes, as long as the excipient is an acceptable excipient in the cosmetic or pharmaceutical field, in accordance with the general knowledge in the art. The excipient content is the content conventional in the art.
Wherein the excipient may be in the form of a water phase, an oil phase, a gel, a wax-in-water emulsion, an oil-in-water emulsion or a water-in-oil emulsion.
The skin external preparation of the present invention may further comprise any other components conventionally used in the cosmetic field, for example, one or more of a preservative, a fragrance, a hydrophilic active agent and a lipophilic active agent. The content of the above-mentioned other components may be a content conventional in the art.
In the present invention, the external skin preparation may further include a particulate phase, and the particulate phase may be one or more of a pigment, a pearlescent agent and a filler.
In the present invention, the form of the external preparation for skin may be any suitable product form including, but not limited to, one or more of aerosol type spray, cream, lotion, solid, liquid, dispersion, foam, gel, lotion, mousse, ointment, powder, patch, pomade, pump spray, stick, mask and towelette. As is well known, the skin external preparation of the present invention may be a cosmetic, dermatological or pharmaceutical topical application product, and the preparation method thereof may be a preparation method conventional in the art.
In a preferred embodiment of the present invention, the external preparation for skin is a cosmetic water, and the external preparation for skin comprises the following components by weight: 5% of glycerin, 3-6% of polyalcohol A, 1% of betaine, 0.5% of panthenol, 0.05-0.1% of dipotassium glycyrrhizinate, 0.15-0.2% of polyethylene glycol-320.5%, 0.15-0.2% of surfactant A, 0.4-0.45% of preservative A, 0-0.15% of xanthan gum, 0.03% of spice and 0.001-10% of nitraria tangutorum bobr extract; the rest is supplemented to 100 percent by deionized water; wherein the polyalcohol A is one or more of butanediol, 1, 2-pentanediol and dipropylene glycol; the surfactant A is polysorbate-20 and/or PEG-40 hydrogenated castor oil; the preservative A is methyl hydroxybenzoate and/or phenoxyethanol.
Wherein, the nitraria tangutorum bobr extract is preferably a nitraria tangutorum bobr seed extract. The content of the Nitraria tangutorum bobr seed extract is preferably 0.0025 to 0.15%, more preferably 0.005 to 0.1%, for example 0.005%, by weight in the skin external preparation.
In a second preferred embodiment of the present invention, the external skin preparation is an essence, and the external skin preparation comprises the following components by weight: 5% of glycerin, 3-6% of polyalcohol B, 1% of panthenol, 0.1% of dipotassium glycyrrhizinate, 0.1% of allantoin, 0.05-3.6% of surfactant B, 0.25-0.45% of preservative B, 0.2-0.4% of xanthan gum, 20.3% of acrylic resin Pemulen TR and 0.05% of spice; and 0.001% -10% of the nitraria tangutorum bobr extract; the rest is supplemented to 100 percent by deionized water; wherein the polyalcohol B is butanediol and/or 1, 3-propanediol; the surfactant B is polyglycerol-2 oleate and/or polyglycerol-10 oleate; the preservative B is one or more of propyl hydroxybenzoate, methyl hydroxybenzoate and phenoxyethanol.
According to the common knowledge in the field, the acrylic resin Pemulen TR-2 is an acrylic acid (ester)/C10-30Alkanol acrylate cross-linked polymers, produced by nuch chemical company, usa.
According to the common knowledge in the art, the xanthan gum is an extracellular microbial polysaccharide produced by fermentation of xanthomonas sp.
Wherein, the nitraria tangutorum bobr extractive is preferably nitraria tangutorum bobr seed extractive. The content of the nitraria tangutorum bobr seed extract is preferably 0.0025 to 0.15%, more preferably 0.005 to 0.1%, for example 0.1%, and the percentage refers to the weight percentage in the skin external preparation.
In a third preferred embodiment of the present invention, the external skin preparation is an emulsion, and the external skin preparation comprises the following components in percentage by weight: 5% of glycerol, 5% of 1, 3-propylene glycol, 1% of panthenol, 0.5% of tocopherol acetate, 2% of polydimethylsiloxane, 1652% of emulsifier A, 2-3% of oil C, 3% of isopropyl isostearate, 0.5% of cetostearyl alcohol, 0.3% of preservative C, 0.5% of xanthan gum and 0.1% of EDTA disodium; the skin external agent also comprises triethanolamine, and the content of the triethanolamine is based on adjusting the pH value of the skin external agent to 5.5-7; 0.001% -10% of nitraria tangutorum bobr extract; the rest is supplemented to 100 percent by deionized water; the oil C is mineral oil and/or isohexadecane; the preservative C is methyl hydroxybenzoate and/or propyl hydroxybenzoate.
The emulsifier A165 is a commercially available product containing glyceryl stearate and PEG-100 stearate, as is common in the art. The hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer is a thickening agent which is produced by Sabic corporation and has the mark of Sepinov EMT 10.
Wherein, the nitraria tangutorum bobr extract is preferably a nitraria tangutorum bobr seed extract. The content of the Nitraria tangutorum bobr seed extract is preferably 0.0025 to 0.15%, more preferably 0.005 to 0.1%, for example 0.005%, by weight in the skin external preparation.
In a fourth preferred embodiment of the present invention, the external preparation for skin is a cream, and the external preparation for skin comprises the following components in percentage by weight: 5% of glycerol, 5% of 1, 3-propylene glycol, 1% of panthenol, 0.5% of tocopherol acetate, 2% of polydimethylsiloxane, 1652% of emulsifier A, 2-8% of oil D, 3% of isopropyl isostearate, 2-3% of cetearyl alcohol, 0.3% of preservative D, 2% of hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and 0.1% of disodium EDTA; the skin external agent also comprises triethanolamine, and the content of the triethanolamine is based on adjusting the pH value of the skin external agent to 5.5-7; 0.001% -10% of nitraria tangutorum bobr extract; the rest is supplemented to 100 percent by deionized water; wherein the oil D is mineral oil and/or isohexadecane; the preservative D is methyl hydroxybenzoate and/or propyl hydroxybenzoate.
Wherein, the nitraria tangutorum bobr extract is preferably a nitraria tangutorum bobr seed extract. The content of the nitraria tangutorum bobr seed extract is preferably 0.0025 to 0.15%, more preferably 0.005 to 0.1%, for example 0.1%, and the percentage refers to the weight percentage in the skin external preparation.
On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows:
(1) the application finds that the Nitraria tangutorum bobr extract, particularly the Nitraria tangutorum bobr seed extract, has the effects of scavenging free radicals, inhibiting tyrosinase activity, inhibiting elastase activity, improving the expression of anti-aging proteins in human fibroblasts, regulating and controlling the synthesis of collagen in dermis, and has the effect of delaying aging, particularly the effects of scavenging free radicals and inhibiting elastase. Compared with a blank control, the generation rates of collagen I, collagen V and BMP1 in the fibroblasts are 136%, 115% and 151% respectively, and the method has important significance for developing skin care products or cosmetics related to anti-aging in the future.
(2) The application discovers for the first time that the nitraria tangutorum bobr extract, particularly the nitraria tangutorum bobr seed extract, can promote the generation of bone morphogenetic protein-1 (BMP-1), and has important significance for the research of the action mechanism of the nitraria tangutorum bobr extract and the active ingredients acting on the BMP-1 target.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
Example 1 preparation of Nitraria seed extract
Crushing the dried nitraria tangutorum bobr seeds, sieving the crushed nitraria tangutorum bobr seeds with a 40-mesh sieve, putting the nitraria tangutorum bobr seeds into a supercritical extraction instrument, and feeding 300g of the nitraria tangutorum bobr seeds at the first feeding amount, wherein the extraction pressure is 35MPa, the extraction temperature is 45 ℃ and CO is used2Flow rate of 25kg/h, supercritical CO2Extracting for 2.5 hours, and collecting the nitraria seed oil from the separation kettle; feeding the materials for 5 times, and extracting 1.5Kg of raw materials to obtain 150g of nitrariatangutorum seed oil. Weighing 10g of Nitraria sibirica seed oil, passing through macroporous adsorption resin AB-8, sequentially eluting with 1 column volume of water and 5 column volumes of 30% ethanol, and discarding the eluate. Then eluting with 70% ethanol with 5 times of column volume, collecting eluate, concentrating, and evaporating to obtain Nitraria tangutorum bobr seed extract dry powder 0.9 g.
Example 2
The toning lotion is prepared from the following components in percentage by weight:
5% of glycerin, 3% of butanediol, 1% of betaine, 0.5% of panthenol, 0.1% of dipotassium glycyrrhizinate, 2.0% of allantoin, 0.15-0.2% of polyethylene glycol-320.5%, 0.15-0.2% of hydroxyethyl cellulose, IS-450.4%, 0.15% of xanthan gum, 0.03% of perfume and 0.005% of nitraria seed extract; the remainder was made up to 100% with deionized water.
The skin external preparation in this example was prepared according to a conventional method for preparing a cosmetic lotion in the art.
Example 3
The essence is prepared from the following components in percentage by weight:
5% of glycerin, 3% of butanediol, 1% of panthenol, 0.1% of dipotassium glycyrrhizinate, 0.1% of allantoin, 4% of nicotinamide, 0.05% of hydroxyethyl cellulose, 0.15% of methylparaben, 0.3% of phenoxyethanol, 0.2% of xanthan gum, 20.3% of acrylic resin Pemulen TR, 0.05% of perfume and 0.1% of nitraria seed extract; the remainder was made up to 100% with deionized water.
The skin external preparation in this example was prepared according to a conventional preparation method of essence in the art.
Example 4
The emulsion is prepared from the following components in percentage by weight:
5% of glycerol, 5% of 1, 3-propylene glycol, 1% of panthenol, 0.5% of tocopherol acetate, 0.1% of dipotassium glycyrrhizinate, 2% of polydimethylsiloxane, 1652% of an emulsifier A, 3% of isopropyl isostearate, 3% of mineral oil, 0.5% of cetostearyl alcohol, 0.2% of methyl hydroxybenzoate, 0.1% of propyl hydroxybenzoate, 0.5% of xanthan gum and 0.1% of disodium EDTA, and regulating the pH value of the emulsion to 5.5-7 by triethanolamine; nitraria seed extract 0.005%; the remainder was made up to 100% with deionized water.
The skin external preparation in this example was prepared according to the conventional emulsion preparation method in the art.
Example 5
The cream is prepared from the following components in percentage by weight:
5% of glycerol, 5% of 1, 3-propylene glycol, 1% of panthenol, 0.5% of tocopherol acetate, 0.1% of dipotassium glycyrrhizinate, 2% of polydimethylsiloxane, 1652% of an emulsifier A, 3% of isopropyl isostearate, 2% of mineral oil, 2% of cetostearyl alcohol, 0.2% of methyl hydroxybenzoate, 0.1% of propyl hydroxybenzoate, 2% of hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and 0.1% of disodium EDTA; regulating the pH value of the emulsion to 5.5-7 by triethanolamine; nitraria seed extract 0.1%; the remainder was made up to 100% with deionized water.
The skin external preparation in this example was prepared according to a conventional method for preparing a cream in the art.
Effect example 1 Activity assay of Nitraria seed extract
(1) Detection of activity of scavenging free radicals by DPPH method
1, 1-Diphenyl-2-trinitrophenylhydrazine (1, 1-Diphenyl-2-piperidinylhydrazyl, DPPH) is a stable nitrogen center organic free radical, and an ethanol solution of the stable nitrogen center organic free radical is dark purple and has strong absorption at about 517 nm; in the presence of free radical scavengers, DPPH absorption is diminished by pairing with its single electron, and gradually disappears. The degree of color fading of the DPPH ethanol solution is in a quantitative relation with the number of electrons received by the DPPH ethanol solution, so that a spectrophotometer can be used for carrying out rapid quantitative analysis, detecting the condition of scavenging free radicals and evaluating the capacity of the antioxidant for scavenging free radicals, the antioxidant capacity and the anti-aging capacity.
Preparing a DPPH ethanol solution: 6mg of DPPH was weighed out accurately, dissolved in 95% ethanol and placed in a 50mL volumetric flask, and dissolved with the aid of ultrasound. When in use, the mixture is diluted four times by 95% ethanol to obtain DPPH ethanol solution with the concentration of 3mg/100mL (76 mu M), and the DPPH ethanol solution is stored in a dark place (0-4 ℃).
Pretreating a sample to be detected: the nitraria tangutorum bobr seed extract prepared in example 1 and an ascorbic acid standard are dissolved in deionized water and diluted to a suitable concentration with the deionized water for later use.
The experimental steps are as follows: adding 2mL of sample solution to be detected into a test tube, then adding 2mL of ethanol solution of LDPPH, and uniformly mixing. Reacting at room temperature in dark for 30min in the dark, and measuring absorbance Abs at 525 nm; except for the sample group, a sample solvent control group, a blank control group and a solvent control group (solvent of DPPH) are respectively arranged, and the positive control group is an ascorbic acid group.
Clearance I (%) ═ 1- (T-T)0)/(C-C0)]×100％
In the formula:
T0: adding the absorbance of the sample solution to be detected with equal volume of 95% ethanol (sample solvent control group);
t: adding the absorbance of an isometric DPPH solution into a sample solution to be detected (sample group);
C0: absorbance of distilled water plus equal volume of 95% ethanol (solvent control);
c: absorbance of distilled water plus an equal volume of DPPH solution (blank control).
TABLE 1 measurement results of radical scavenging action
As can be seen from Table 1, the Nitraria tangutorum bobr seed extract has DPPH radical scavenging effect. Compared with the blank control group, the radical clearance rate of the nitraria seed extract group is 88%.
(2) Detection of tyrosinase Activity inhibition Rate
The experimental steps are as follows: adding phosphate buffer solution (pH6.8, 30mM)70 μ L, 1.66mM tyrosinase solution 80 μ L, and sample solution 80 μ L into 96-well plate, mixing, incubating at 37 deg.C for more than 5 min, adding tyrosinase solution (125U/ml) (from Sigma, product number T3824)10 μ L, incubating at 37 deg.C for 10min, and measuring absorbance A at 475nm wavelength with enzyme-labeling instrument475The sample solution includes the solution prepared from the nitraria tangutorum bobr seed extract prepared in example 1 and an arbutin standard solution (both diluted to a suitable concentration by deionization), wherein the arbutin standard solution is used as a positive control group. The deionized water is used for replacing the sample water solution, the absorbance is also measured as a reference solution, and the tyrosinase activity inhibition rate calculation formula is as follows: inhibition ratio (%) (A)0-(A475-B))/A0X 100% where A0Is the absorbance of a reference solution, A475Is the absorbance of the sample solution, and B is the absorbance of the sample blank solution.
TABLE 2 determination of tyrosinase inhibitory Activity
As can be seen from Table 2, the Nitraria tangutorum bobr seed extract has tyrosinase inhibitory effect. Compared with a blank control group, the tyrosinase inhibition rate of the nitraria seed extract group is 21%.
(3) Detection of Elastase Activity inhibition
The experimental steps are as follows: mu.L of the sample solution and 130. mu.L of 0.1M Tris-HCl buffer solution (pH8.0) containing 1.015mM of the reaction substrate Succ-Ala-Ala-Ala-p-nitroanilide were added to a 96-well plate, incubated at 25 ℃ for 5 minutes, 15. mu.L of elastase solution (0.5U/mL) was added, and the incubation continued at 25 ℃Incubation for 30min, followed by determination of the absorbance A at a wavelength of 410nm using a microplate reader410The sample solution includes the prepared solution of the Nitraria seed extract prepared in example 1 and an Elastatin solution (both diluted to appropriate concentrations by deionization), wherein the Elastatin solution (elastase inhibitor, available from sigma, Inc., under the code E0881) is used as a positive control group. The absorbance was also measured as a reference solution by replacing the sample aqueous solution with deionized water.
The calculation formula of the elastase activity inhibition rate is as follows: inhibition ratio (%) (A)0-(A410-B))/A0X 100% where A0Is the absorbance of a reference solution, A410Is the absorbance of the sample solution, and B is the absorbance of the sample blank solution.
TABLE 3 measurement results of elastase activity inhibition
As is clear from Table 3, Nitraria tangutorum bobr seed extract has elastase inhibitory activity. Compared with the blank control group, the elastase inhibition rate of the nitraria tangutorum bobr seed extract is 22%.
(4) Fibroblast collagen I production rate assay
Human fibroblasts were seeded in a 96-well plate (5000 cells/well), the culture medium was removed after 72 hours of culture, the solution prepared from the Nitraria seed extract prepared in example 1 and an ascorbic acid standard solution (both diluted to appropriate concentrations with deionization) were added, and further cultured for 48 hours, and after removal of the supernatant, the COL I content in the cells was determined by immunofluorescence assay, and the DNA content in the cell layer (picogreen kit) was also determined for COL I content data normalization. The test results will indicate that the blank control group had 100% production of type I stock collagen compared to the untested control group. The formation rate is more than 100%, which is the promotion effect.
TABLE 4 COL I content measurement results
As can be seen from table 4, the nitraria tangutorum bobr seed extract has the effect of promoting collagen I secretion. The production rate of COL I in the human fibroblasts of the nitraria seed extract-treated group was 136% compared to that of the blank control group.
(5) Fibroblast collagen V production rate assay
Human fibroblasts were seeded in a 96-well plate (5000 cells/well), the culture medium was removed after 72 hours of culture, a solution prepared from the Nitraria seed extract prepared in example 1 and an ascorbic acid standard solution (both diluted to appropriate concentrations with deionized water) were added, and further culture was carried out for 48 hours, after removal of the supernatant, the COL V content in the cells was measured by the immunofluorescence assay, and the DNA content of the cell layer (picogreen kit) was also measured for COL V content data normalization. The test result shows that the generation rate of V-type stored collagen of the blank control group is 100 percent compared with the control group which is not tested. The formation rate is more than 100%, which is the promotion effect.
TABLE 5 COL V content measurement results
As can be seen from table 5, the nitraria seed extract had the effect of promoting collagen V secretion, and the production rate of COL V in human fibroblasts of the nitraria seed extract-treated group was 115% compared to that of the blank control group.
(6) Fibroblast BMP1 production Rate determination
Inoculating human fibroblasts into a 96-well plate (5000 cells/well), culturing for 72 hours, removing a culture medium, adding a solution prepared from the nitraria tangutorum bobr seed extract prepared in example 1 (diluted to a proper concentration by deionization), culturing for 24 hours, collecting cell culture supernatant, and measuring the content of BMP1 in the cell culture supernatant by adopting a BMP1ELISA kit; the BMP1 content data were normalized by simultaneously determining the DNA content of the cell layer (picogreen kit, from thermo fisher). The test results showed that the generation rate of BMP1 in the blank control group was 100% compared with the control group not tested. The formation rate is more than 100%, which is the promotion effect.
TABLE 6 measurement of BMP1 content
As can be seen from Table 6, the Nitraria tangutorum bobr seed extract has the effect of promoting BMP 1. Compared with a blank control group, the generation rate of BMP1 in the human fibroblasts of the nitraria seed extract-treated group can reach 151%.
Claims (11)
1. Use of Nitraria tangutorum bobr extract in preparing elastic fiber hydrolysis inhibitor and/or skin aging resisting protein expression up-regulator;
(1) the elastic fiber hydrolysis inhibitor is an elastase inhibitor;
the skin aging resistant protein expression up-regulator is one or more of BMP-1 expression up-regulator, collagen I expression up-regulator and collagen V expression up-regulator;
(2) the preparation method of the nitraria tangutorum bobr extract comprises the following steps: mixing nitraria tangutorum bobr with supercritical CO2Mixing, and extracting to obtain extractive solution; wherein:
the extraction pressure is 35-40 MPa;
the supercritical CO2The flow rate of the water is 20-30 kg/h;
purifying the extract liquor, and carrying out macroporous adsorption resin chromatography on the extract liquor; the kind of the macroporous adsorption resin is AB-8; the eluent for chromatography is ethanol water solution, the concentration of the ethanol water solution is 65-75%, and the percentage refers to volume percentage;
and water elution impurity removal treatment is carried out before chromatography, and after the water elution impurity removal, 25-35% ethanol water solution with 5 times of column volume is used for elution impurity removal, wherein the percentage refers to volume percentage.
2. The use according to claim 1,
when the Nitraria tangutorum bobr extract is used for preparing the elastic fiber hydrolysis inhibitor, the concentration of the Nitraria tangutorum bobr extract in the elastic fiber hydrolysis inhibitor is 0.5-1.5 mg/mL;
when the nitraria tangutorum bobr extract is used for preparing the skin aging resistance protein expression up-regulator, the concentration of the nitraria tangutorum bobr extract in the skin aging resistance protein expression up-regulator is 0.05-1 mg/mL.
3. The use according to claim 2,
when the Nitraria tangutorum bobr extract is used for preparing the elastic fiber hydrolysis inhibitor, the concentration of the Nitraria tangutorum bobr extract in the elastic fiber hydrolysis inhibitor is 1 mg/mL;
when the anti-skin aging protein is BMP-1, the concentration of the nitraria tangutorum bobr extract in the up-regulator for the expression of the anti-skin aging protein is 0.05 mg/mL;
when the anti-skin aging protein is collagen I or collagen V, the concentration of the nitraria tangutorum bobr extract in the anti-skin aging protein expression up-regulator is 1 mg/mL.
4. The use of claim 1, wherein the extraction site of Nitraria tangutorum bobr is Nitraria tangutorum bobr seed;
and/or, pretreating the nitraria tangutorum bobr;
and/or the pressure of the extraction is 35 MPa;
and/or the temperature of the extraction is 45 ℃;
and/or the supercritical CO2The flow rate of (2) is 25 kg/h;
and/or the extraction time is 2-3 h;
and/or the concentration of the ethanol water solution is 70%, and the percentage refers to volume percentage.
5. The use according to claim 4, wherein the pre-treatment is a comminution treatment.
6. The use according to claim 4, wherein the extraction time is 2.5 h.
7. The use of claim 4, wherein the volume of eluent for chromatography is 4 to 6 column volumes.
8. The use of claim 7, wherein the volume of eluent for chromatography is 5 column volumes.
9. The use of claim 4, wherein the water elutes in a volume of 1 column volume.
10. The use according to claim 5, wherein the comminution is followed by a sieving treatment.
11. The use according to claim 10, wherein the screened mesh size is 40 mesh.
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| PCT/CN2019/096712 WO2020015726A1 (en) | 2018-07-20 | 2019-07-19 | Use of nitraria tangutorum bobr. extracts and skin external preparations comprising the same |
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| JP4781062B2 (en) * | 2005-09-14 | 2011-09-28 | 株式会社ノエビア | Skin preparation for moisturizing or improving rough skin |
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| CN101991641B (en) * | 2010-11-09 | 2011-11-02 | 中国科学院西北高原生物研究所 | Preparation method of antioxidation nitraria tangutorum bobr extractive |
| CN105232593A (en) * | 2015-10-28 | 2016-01-13 | 青海尤尼克生物技术有限公司 | Nitraria branch and leaf extract |
| CN105250182A (en) * | 2015-11-17 | 2016-01-20 | 中国科学院西北高原生物研究所 | Skin care composition containing nitraria tangutorum extract |
| CN108379311A (en) * | 2018-05-16 | 2018-08-10 | 中国科学院新疆理化技术研究所 | A kind of preparation method and applications of Nitraria sibirica leaves extract |
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| Title |
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| JAT RANA et al..Total Antioxidant Capacity and Characterization of Nitraria tangutorum Fruit Extract by Rapid Bioassay-Directed Fractionation.《Journal of AOAC International》.2016,第99卷(第5期),第1219-1222页. * |
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