CN110731919B - Application of mirabilis jalapa extract and skin external preparation containing mirabilis jalapa extract - Google Patents
Application of mirabilis jalapa extract and skin external preparation containing mirabilis jalapa extract Download PDFInfo
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- CN110731919B CN110731919B CN201810804107.XA CN201810804107A CN110731919B CN 110731919 B CN110731919 B CN 110731919B CN 201810804107 A CN201810804107 A CN 201810804107A CN 110731919 B CN110731919 B CN 110731919B
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- mirabilis jalapa
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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
- 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
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 mirabilis jalapa extract and a skin external preparation containing the same. The application specifically is the application of the mirabilis jalapa 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 Mirabilis jalapa extract, especially Mirabilis jalapa extract, has effects of scavenging free radical activity, inhibiting tyrosinase activity, inhibiting elastase activity, improving anti-aging protein expression in human fibroblast, regulating synthesis of collagen in dermis, and delaying aging.
Description
Technical Field
The invention relates to application of a mirabilis jalapa extract and a skin external preparation containing the same.
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 two forms of intrinsic aging and extrinsic aging. Intrinsic aging is natural aging, which is 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 skin laxity. 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. In short, BMP-1 functionalizes collagen, ensuring that collagen is processed and modified in the desired 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. In the case of skin, collagen is a light propagation channel, and collagen fiber-mediated light scattering plays an important role in light propagation penetration inside 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 the 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.
Mirabilis jalapa L.is a plant of Mirabilis, also known as nopalina, Sophora fargesii, Nostoc sphaeroides, Sophora japonica, Eugenia caryophyllata, and Eugenia caryophyllata. The native tropical America is cultivated in various places in China, namely south and north, and is an ornamental flower and sometimes a wild flower. The medicine is originally seen in Bencao gang mu Shi Yi (compendium of materia Medica), is cold in nature, sweet in taste, applicable to all kinds of medicines such as root, leaf, flower and seed, and can be used for treating symptoms such as heat stranguria, whitish and turbid urine, edema, leucorrhea with reddish discharge, joint swelling and pain, carbuncle, sore and pyogenic infections, and the seed powdery mildew can remove facial freckles and nevus. The research reports that the water extract of the mirabilis jalapa contains amino acids, saccharides, glycosides, organic acids, phenols, volatile oil and other substances, the alcohol extract contains phenols, cardiac glycosides, lactones, coumarins, flavones and other substances, and the petroleum ether extract mainly contains saponins and other substances.
Disclosure of Invention
The invention provides application of a mirabilis jalapa extract and a skin external preparation containing the mirabilis jalapa 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 invention discovers that the mirabilis jalapa extract has a definite action mechanism, is 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) mirabilis jalapa extract has free radical scavenging effect; 2) mirabilis jalapa extract has tyrosinase activity inhibiting effect; 3) the Mirabilis jalapa extract has effects of inhibiting elastase activity, improving anti-aging protein expression in human fibroblast, regulating synthesis of collagen in dermis, and delaying aging, and is suitable for cosmetics.
The invention mainly solves the technical problems by the following technical means:
the invention provides an application of a mirabilis jalapa 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 trap and neutralize free radicals, such as a free radical scavenger, of a type that 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 skin aging-resistant protein expression up-regulator may be a substance conventionally used in the art to affect the expression of skin aging-related proteins, 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 invention, the concentration of the mirabilis jalapa 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 mirabilis jalapa extract in the melanin synthesis inhibitor is preferably 0.25-0.75 mg/mL, and more preferably 0.5 mg/mL.
In the present invention, the concentration of the mirabilis jalapa extract in the elastic fiber hydrolysis inhibitor is preferably 0.5-1.5 mg/mL, and more preferably 1 mg/mL.
In the invention, the concentration of the mirabilis jalapa extract in the skin aging resistant protein expression up-regulator 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 Mirabilis jalapa 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 mirabilis jalapa extract in the up-regulator of anti-skin aging protein expression 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 invention, the Mirabilis jalapa extract can be prepared by a conventional method in the field, for example, the Mirabilis jalapa L is mixed with a solvent and extracted to obtain an extracting solution.
The extract part of Mirabilis jalapa can be any part of Mirabilis jalapa which can be used as medicine, such as one or more of Mirabilis jalapa root, Mirabilis jalapa leaf, Mirabilis jalapa flower and Mirabilis jalapa seed, preferably Mirabilis jalapa flower. The Mirabilis jalapa L is obtained by collecting Mirabilis jalapa L in bud stage.
When the extracting part of the mirabilis jalapa is mirabilis jalapa, the mirabilis jalapa extract is a mirabilis jalapa extract.
Wherein, the solvent can be an extraction solvent which is conventional in the field, and preferably ethanol. The concentration of the ethanol can be the conventional concentration in the field, and is preferably 60-80% ethanol water solution, and the percentage refers to volume percentage.
Wherein, the ratio of the quality of the mirabilis jalapa to the volume of the solvent can be a conventional ratio in the field, such as 1: (8-12) g/mL, preferably 1: 10 g/mL.
The extraction method can be conventional extraction method in the field, such as cold soaking extraction and heating extraction in sequence.
The time for the cold leaching extraction may be a time conventional in the art, for example 12 h.
The temperature for the heating extraction may be a temperature conventional in the art, for example, 50 to 70 ℃, preferably 60 ℃.
The time for the heating extraction can be a time conventional in the art, such as 1-3 h, preferably 2 h.
The number of times of the heating extraction may be a number of times conventional in the art, for example, 3 times. It is known to those skilled in the art that when the number of extraction is more than 1, the extracts should be combined.
Wherein the extract can be purified by conventional procedures in the art, for example, subjecting the extract to macroporous adsorbent resin chromatography.
Preferably, the extract is dried, redissolved with water and chromatographed on macroporous adsorbent resin.
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 25 to 35%, more preferably 30%, and the percentage refers to volume percentage.
The volume of eluent for the chromatography may be 5 column volumes.
The chromatography may also be preceded by a depuration treatment, for example by elution with water, the elution volume of which is preferably 1 column volume.
Preferably, the preparation method of the mirabilis jalapa extract comprises the following steps: mixing flos Mirabilis with ethanol water solution, sequentially cold soaking and heating for extraction to obtain extractive solution; the concentration of the ethanol water solution is 60-80%, and the percentage refers to volume percentage; the cold leaching extraction time is 12 hours; the temperature for heating and extracting is 50-70 ℃; the heating extraction time is 1-3 h.
The invention also provides application of the mirabilis jalapa extract as an antioxidant active ingredient, a whitening active ingredient and an anti-aging active ingredient in cosmetics or skin care products.
Wherein the Mirabilis jalapa extract is as defined above.
The invention also provides a skin external preparation, which contains 0.001-10% of mirabilis jalapa extract, wherein the percentage refers to the weight percentage (g/100mL) in the skin external preparation.
Wherein the Mirabilis jalapa extract is as defined above.
Wherein, the content of the mirabilis jalapa extract is preferably 0.0025-0.15%, such as 0.005%, 0.05% or 0.1%, and the percentage refers to the weight percentage in the skin external preparation.
When the mirabilis jalapa extract is used as an antioxidant in the skin external preparation, the content of the mirabilis jalapa extract is preferably 0.0025 to 0.0075%, more preferably 0.005%, and the percentage refers to the weight percentage in the skin external preparation.
When the mirabilis jalapa extract is used as a melanin synthesis inhibitor in the skin external preparation, the content of the mirabilis jalapa extract is preferably 0.025-0.075%, more preferably 0.05%, and the percentage refers to the weight percentage in the skin external preparation.
When the mirabilis jalapa extract is used as a skin elastic fiber hydrolysis inhibitor in the skin external preparation, the content of the mirabilis jalapa extract is preferably 0.05-0.15%, more preferably 0.1%, and the percentage refers to the weight percentage in the skin external preparation.
When the mirabilis jalapa extract is used as an up-regulator of expression of anti-skin aging proteins in the external preparation for skin, the content of the mirabilis jalapa extract is preferably 0.005-0.1%, for example 0.005% or 0.1%, and the percentage refers to the weight percentage in the external preparation for skin.
When the anti-skin aging protein is BMP-1, the content of the mirabilis jalapa extract is preferably 0.005%, and the percentage refers to the weight percentage in the external preparation for skin.
When the anti-skin aging protein is collagen I or collagen V, the content of the mirabilis jalapa extract is preferably 0.1%, by weight, in the skin external preparation.
In the present invention, the external preparation for skin refers to a general concept of all ingredients generally used for the external skin, and may be, for example, cosmetics or medicines. The cosmetic can be basic cosmetics, face makeup cosmetics, body makeup cosmetics, head care products and the like, and the dosage form of the cosmetic is not particularly limited and can be reasonably selected according to different purposes.
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 mirabilis jalapa 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 one or more of polysorbate-20 and PEG-40 hydrogenated castor oil; the preservative A is methyl hydroxybenzoate and/or phenoxyethanol.
Wherein, the mirabilis jalapa extract is preferably a mirabilis jalapa extract. The content of the mirabilis jalapa flower 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 glycerol, 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 mirabilis jalapa 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)/C10-30Alkyl alcohol acrylic acidEster cross-linked polymers, produced by the U.S. Noyu chemical company.
According to the common knowledge in the art, the xanthan gum is an extracellular microbial polysaccharide produced by fermentation of xanthomonas sp.
Wherein, the mirabilis jalapa extract is preferably a mirabilis jalapa extract. The content of the mirabilis jalapa flower 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 preparation for skin is an emulsion, 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, 2-3% 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 triethanolamine is contained in an amount to adjust the pH value of the skin external agent to 5.5-7; and 0.001% -10% of the mirabilis jalapa extract; the rest is supplemented to 100 percent by deionized water; wherein 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 mirabilis jalapa extract is preferably a mirabilis jalapa extract. The content of the mirabilis jalapa flower 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 cetostearyl 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 mirabilis jalapa 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 Mirabilis jalapa extract is preferably Mirabilis jalapa extract. The content of the mirabilis jalapa flower 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 discovers that the mirabilis jalapa extract, particularly the mirabilis jalapa extract, has the effects of eliminating free radical activity, inhibiting tyrosinase activity, inhibiting elastase activity and improving anti-aging protein expression in human fibroblasts, can regulate and control synthesis of collagen in dermis, has the effect of delaying aging, and particularly has the effect of improving anti-aging protein expression in human fibroblasts. Compared with a blank control, the generation rates of collagen I, collagen V and BMP1 in the fibroblasts are 133%, 125% and 150% 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 mirabilis jalapa extract, particularly the mirabilis jalapa extract, can promote the generation of bone morphogenetic protein-1 (BMP-1), and has important significance for the research on the action mechanism of the mirabilis jalapa 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 Mirabilis jalapa extract
Weighing 100g of dried mirabilis jalapa, crushing, adding 100mL of 70% ethanol, and cold soaking for 12 hours until the solvent fully soaks the material. Heating to 60 deg.C, extracting for 2 hr, filtering, collecting extractive solution, and extracting for 3 times. Mixing the three extractive solutions, and vacuum drying under reduced pressure to obtain 15g crude extract. Weighing 1g of crude extract, dissolving with water, passing through macroporous adsorbent resin AB-8, eluting with 1 column volume of water, and discarding water eluate. Then eluting with 30% ethanol with 5 times of column volume, concentrating eluent, and evaporating to obtain Mirabilis jalapa flower extract dry powder 0.5 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, 450.4% of IS-xanthan gum, 0.03% of perfume and 0.005% of mirabilis jalapa 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 mirabilis jalapa 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; mirabilis jalapa flower extract 0.005%; the remainder was made up to 100% with deionized water.
The skin preparation for external use in this example was prepared according to a method for preparing an emulsion that is conventional 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; mirabilis jalapa flower 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 mirabilis jalapa 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 by four times by 95% ethanol to obtain a 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 mirabilis jalapa flower 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 in dark place at room temperature for 30min in the dark place in a dark place, 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 the sample solution to be detected and an isopyknic DPPH solution (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 of radical scavenging action
As shown in Table 1, Mirabilis jalapa extract has DPPH radical scavenging effect. Compared with a blank control group, the radical clearance rate of the mirabilis jalapa extract group is 76%.
(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 a solution prepared from the mirabilis jalapa 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 (%) ═ a0-(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, Mirabilis jalapa flower extract has tyrosinase inhibitory activity. Compared with a blank control group, the tyrosinase inhibition rate of the mirabilis jalapa extract group is 44%.
(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 an elastase solution (0.5U/mL) was added, incubation at 25 ℃ was continued for 30 minutes, and then the absorbance A at 410nm was measured with a microplate reader410The sample solution includes a prepared solution of the mirabilis jalapa extract prepared in example 1 and an Elastatinal solution (both diluted to an appropriate concentration by deionization), wherein the Elastatinal solution (elastase inhibitor, available from sigma, cat # 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 shown in Table 3, Mirabilis jalapa flower extract had an elastase inhibitory effect. The inhibition rate of elastase in the mirabilis jalapa extract group is 26% compared with that of the blank control group.
(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, a solution prepared from the mirabilis jalapa 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 the supernatant was removed, the content of COL I (collagen I) in the cells was measured by the immunofluorescence assay, and at the same time, the DNA content of the cell layer (picogreen kit, purchased from thermo fisher) was measured to standardize the content data of COL I. 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 mirabilis jalapa extract had the effect of promoting collagen I secretion, and the COL I production rate in human fibroblasts of the mirabilis jalapa extract-treated group was 133% compared to 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 mirabilis jalapa 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 the supernatant was removed, the content of COL V (collagen V) in the cells was measured by the cytoimmunofluorescence method, and at the same time, the DNA content of the cell layer (picogreen kit, purchased from thermo fisher) was measured to standardize the content data of COL V. 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 measurement results of COL V content
As can be seen from table 5, the mirabilis jalapa extract has the effect of promoting collagen V secretion, and compared with the blank control group, the percentage of COL V production in human fibroblasts of the mirabilis jalapa extract-treated group can reach 125%.
(6) Fibroblast BMP1 production Rate determination
Planting human fibroblasts into a 96-well plate (5000 cells/well), removing a culture medium after culturing for 72 hours, adding a solution prepared from the mirabilis jalapa 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 100% production of BMP1 in the blank control compared to the control that was not tested. The formation rate is more than 100%, which is the promotion effect.
TABLE 6 measurement of BMP1 content
As shown in Table 6, Mirabilis jalapa extract has an effect of promoting BMP 1. Compared with a blank control group, the generation rate of BMP1 in the human fibroblasts of the Mirabilis jalapa extract treatment group can reach 150%.
Claims (15)
1. Application of Mirabilis jalapa extract in preparing elastic fiber hydrolysis inhibitor and/or skin aging resisting protein expression up-regulating agent;
(1) the preparation method of the mirabilis jalapa extract comprises the following steps: mixing Mirabilis jalapa flower with solvent, and extracting to obtain extractive solution;
the solvent is ethanol water solution, the concentration of the ethanol water solution is 60-80%, and the percentage refers to volume percentage;
the extract is further subjected to purification treatment: drying the extracting solution, redissolving with water, and performing macroporous adsorption resin chromatography; 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 25-35%, and the percentage refers to volume percentage;
(2) 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 use according to claim 1,
when the mirabilis jalapa extract is used for preparing the elastic fiber hydrolysis inhibitor, the concentration of the mirabilis jalapa extract in the elastic fiber hydrolysis inhibitor is 0.5-1.5 mg/mL;
when the mirabilis jalapa extract is used for preparing the skin aging resistant protein expression up-regulator, the concentration of the mirabilis jalapa extract in the skin aging resistant protein expression up-regulator is 0.05-1 mg/mL.
3. The use according to claim 2,
when the Mirabilis jalapa extract is used for preparing an elastic fiber hydrolysis inhibitor, the concentration of the Mirabilis jalapa extract in the elastic fiber hydrolysis inhibitor is 1 mg/mL;
when the anti-skin aging protein is BMP-1, the concentration of the mirabilis jalapa extract in the up-regulator of the anti-skin aging protein expression is 0.05 mg/mL;
when the anti-skin aging protein is collagen I or collagen V, the concentration of the mirabilis jalapa extract in the anti-skin aging protein expression up-regulator is 1 mg/mL.
4. The use of claim 1, wherein the ratio of the mass of Mirabilis jalapa to the volume of the solvent is 1: (8-12) g/mL;
and/or the extraction method comprises the steps of cold soaking extraction and heating extraction in sequence;
and/or the eluent for chromatography is ethanol water solution, the concentration of the ethanol water solution is 30%, and the percentage refers to volume percentage.
5. The use of claim 4, wherein the ratio of the mass of Mirabilis jalapa to the volume of the solvent is 1: 10 g/mL.
6. The use according to claim 4, wherein the cold leaching extraction time is 12 h.
7. The use according to claim 4, wherein the temperature of the heated extraction is 50 to 70 ℃.
8. The use according to claim 7, wherein the temperature of the heated extraction is 60 ℃.
9. The use according to claim 4, wherein the time of the heating extraction is 1 to 3 hours.
10. The use according to claim 9, wherein the time of the heating extraction is 2 h.
11. The use of claim 4, wherein the number of heat extractions is 3.
12. The use of claim 4, wherein the volume of eluent for chromatography is 5 column volumes.
13. The use of claim 4, wherein the chromatography is further preceded by a depuration treatment.
14. The use as claimed in claim 13, wherein the chromatography is further preceded by an elution with water to remove impurities.
15. The use of claim 14, wherein the water elutes in a volume of 1 column volume.
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| PCT/CN2019/096711 WO2020015725A1 (en) | 2018-07-20 | 2019-07-19 | Use of mirabilis jalapa l. extracts and skin external preparations comprising the same |
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| CN115177566A (en) * | 2022-07-28 | 2022-10-14 | 杭州舒彩网络科技有限公司 | Synergistic hydroxy pinacolone retinoic acid ester anti-aging essence and preparation method thereof |
| CN115444802B (en) * | 2022-09-05 | 2023-06-23 | 福州萍晴日用化妆品有限公司 | Jasmine moisturizing essence emulsion composition and preparation method thereof |
| CN115400058A (en) * | 2022-09-20 | 2022-11-29 | 杭州舒彩网络科技有限公司 | Composition for improving skin cell activity and application of composition in skin external product |
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| Jyotchna Gogoi, et al..Isolation and characterization of bioactive components from Mirabilis jalapa L.radix.《Journal of Traditional and Complementary Medicine》.2016,第6卷(第1期),第41-47页. * |
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