CN113041180A

CN113041180A - Cosmetic liquid, preparation method thereof and collagenase inhibitor

Info

Publication number: CN113041180A
Application number: CN201911378663.6A
Authority: CN
Inventors: 杨登亮; 翟裕诗; 李传茂; 刘广健; 张楚标; 曾伟丹; 张伟杰
Original assignee: GUANGZHOU BAIYUN LIANJIA FINE CHEMICAL FACTORY; Guangdong Danz Group Co Ltd; Guangzhou Keneng Cosmetic Research Co Ltd
Current assignee: GUANGZHOU BAIYUN LIANJIA FINE CHEMICAL FACTORY; Guangdong Danz Group Co Ltd; Guangzhou Keneng Cosmetic Research Co Ltd
Priority date: 2019-12-27
Filing date: 2019-12-27
Publication date: 2021-06-29

Abstract

The invention provides a beauty lotion, a preparation method thereof and a collagenase inhibitor. The cosmetic liquid comprises a collagenase inhibitor and a penetration enhancer; the addition amount of the collagenase inhibitor is 0.01-10% of the total mass of the cosmetic liquid; the addition amount of the penetration enhancer is 0.01-10%; the collagenase inhibitor comprises a pomegranate rind extract and a chamomile extract, and the addition amount of the pomegranate rind extract is 28-72% and the addition amount of the chamomile extract is 28-72% based on the total mass of the collagenase inhibitor. The beauty lotion has mild formula, and can effectively improve the skin elasticity, thereby playing the role of anti-aging.

Description

Cosmetic liquid, preparation method thereof and collagenase inhibitor

Technical Field

The invention relates to a beauty fluid, a preparation method thereof and a collagenase inhibitor, belonging to the field of cosmetics.

Background

Collagen is mainly produced by fibroblasts in the dermis layer, is a main component of the dermis layer, and can maintain the structure of the skin and endow the skin with toughness and elasticity. The collagen content and distribution determine the youth or not of the skin. However, abnormal reduction of collagen content causes thinning of the dermis, skin sagging, loss of elasticity, appearance of wrinkles, and affects the quality of life of people. With the ongoing and intensive research on collagen, researchers have found that collagenase plays an important role in the dynamic balance of skin collagen, and its overexpression and abnormal activation are one of the major causes of skin aging. Therefore, inhibiting the activity of collagenase can block the degradation of collagen of skin, increase the content of collagen, and realize the anti-aging effect.

The main approaches to skin anti-aging and skin care include the following aspects, with respect to factors affecting the collagen content of the skin: (1) increasing collagen synthesis by stimulating proliferation of fibroblasts in the dermis layer; (2) increasing the total amount of collagen in the dermis by stimulating the speed of synthesizing collagen by fibroblasts through active factors; (3) the degradation speed of the collagen is slowed down by inhibiting the catalytic activity of collagenase, a key enzyme for degrading the collagen in the dermis, so that the anti-aging purpose is achieved; (4) through sun protection, the damage of ultraviolet rays in sunlight to collagen in the skin is prevented, and the photoaging of the skin is slowed down; (5) the induced expression of collagenase and abnormal cross-linking of biomacromolecules is slowed down by scavenging excess oxygen free radicals in the skin using antioxidants.

In order to prevent skin from sagging, losing elasticity, wrinkling, etc., and keep skin young, anti-aging products mixed with various components such as hydrolyzed collagen, hyaluronic acid, polypeptide, retinol and its derivatives have been proposed in the prior art. However, if these ingredients are used in large amounts, problems arise in the actual anti-aging effect, the feeling of use, and safety. If the molecular weight of the hydrolyzed collagen is too large, the hydrolyzed collagen is not easy to permeate the skin barrier of the human body to reach the dermis; hyaluronic acid cannot essentially slow down the loss of collagen; the polypeptide and the retinol have certain irritation and safety risks to the skin, and the like.

With the increase of attention of people to skin health, the development of a natural anti-aging agent with safety, stability, obvious effect and high cost performance has become one of the main research directions of the current pharmaceutical and cosmetic industries, and has a very good development prospect.

Disclosure of Invention

Problems to be solved by the invention

In view of the prior art, the hydrolyzed collagen has too high molecular weight and is not easy to reach the dermis layer through the skin barrier of the human body; hyaluronic acid cannot essentially slow down the loss of collagen; the invention provides a cosmetic liquid, which comprises a collagenase inhibitor and has excellent anti-aging effect.

Further, the present invention provides a collagenase inhibitor capable of inhibiting collagenase activity.

Furthermore, the invention also provides a preparation method of the beauty lotion, which is simple to operate and easy to obtain raw materials.

Means for solving the problems

The invention provides a cosmetic liquid, which comprises a collagenase inhibitor and a penetration enhancer; the addition amount of the collagenase inhibitor is 0.01-10% of the total mass of the cosmetic liquid; the addition amount of the penetration enhancer is 0.01-10%;

the collagenase inhibitor comprises a pomegranate rind extract and a chamomile extract, and the addition amount of the pomegranate rind extract is 28-72% and the addition amount of the chamomile extract is 28-72% based on the total mass of the collagenase inhibitor.

The cosmetic liquid according to the present invention is characterized in that the mass ratio of the pomegranate bark extract to the chamomile extract is 1: 0.39-2.5, preferably 1: 0.4-2.4, more preferably 1: 0.42-2.2, further preferably 1: 0.45-2.1, further preferably 1: 0.5-2, and further preferably 1: 0.55-1.8.

The cosmetic liquid according to the present invention, wherein the penetration enhancer comprises one or a combination of two or more of propylene glycol, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate and chitosan.

The cosmetic liquid according to the present invention, wherein the collagenase is interstitial collagenase.

The beauty fluid further comprises one or more of a humectant, an emulsifier, a pH regulator, a preservative, a skin conditioner, a skin whitening agent, a soothing agent, an antioxidant, a chelating agent and an aromatic;

preferably, the addition amount of the humectant is 0.01-20% based on the total mass of the cosmetic liquid; the addition amount of the emulsifier is 0.01-0.5%; the addition amount of the pH regulator is 0.01-1%; the addition amount of the preservative is 0.01-1.5%; the addition amount of the skin conditioner is 0.01-5%; the addition amount of the skin whitening agent is 0.01-5%; the addition amount of the allergy relieving agent is 0.01-5%; the addition amount of the antioxidant is 0-2%, and the addition amount of the chelating agent is 0.01-1%; the addition amount of the aromatic is 0.01-1%.

The skin conditioner according to the present invention comprises one or a combination of two or more of allantoin, an ulva extract, a phaeodactylum tricornutum extract, a macroalgae extract, a chlorella fermentation product, hydrolyzed collagen, a brown algae extract, a mistletoe ginkgo extract, a bofenate spirulina extract, a cogongrass rhizome extract, and a cactus extract.

The beauty fluid comprises one or more of niacinamide, magnolia sieboldii extract, kojic acid and derivatives thereof, arbutin and derivatives thereof and vitamin C; and/or the presence of a gas in the gas,

the soothing agent comprises one or more of dipotassium glycyrrhizinate, bisabolol, ginger root extract, aloe vera extract, centella asiatica extract, evening primrose oil and folium andrographis paniculatae extract.

The present invention also provides a collagenase inhibitor comprising: the collagenase inhibitor comprises a pomegranate bark extract and a chamomile extract, wherein the addition amount of the pomegranate bark extract is 28-72% and the addition amount of the chamomile extract is 28-72% based on the total mass of the collagenase inhibitor.

The collagenase inhibitor according to the present invention, wherein the mass ratio of the pomegranate bark extract to the chamomile extract is 1: 0.39-2.5, preferably 1: 0.4-2.4, more preferably 1: 0.42-2.2, further preferably 1: 0.45-2.1, further preferably 1: 0.5-2, and further preferably 1: 0.55-1.8.

The present invention further provides a method for preparing the collagenase inhibitor according to the present invention, which comprises the step of mixing the components of the collagenase inhibitor.

ADVANTAGEOUS EFFECTS OF INVENTION

The beauty lotion has mild formula, and can effectively improve the skin elasticity, thereby playing the role of anti-aging.

The collagenase inhibitor of the present invention has an excellent inhibitory effect on collagenase activity and has no side effects on the human body.

The preparation method of the beauty lotion is simple to operate, the raw materials are easy to obtain, and the beauty lotion can be produced in batches.

Drawings

FIG. 1 is a graph showing the logarithmic mass concentration-collagenase activity inhibition ratio of the pomegranate rind extract according to comparative examples 1 to 5 of the present invention;

FIG. 2 is a graph showing the logarithmic mass concentration-collagenase activity inhibition ratios of the chamomile extracts of comparative examples 6 to 10 according to the invention;

FIG. 3 is a graph showing the log mass concentration of collagenase inhibitors versus the collagenase activity inhibition rate for examples 1-5 of the present invention;

FIG. 4 is a graph showing the relationship between the content of pomegranate rind extract and the interaction coefficient in collagenase inhibitors according to examples 1 to 5 of the present invention.

FIG. 5 is a graph showing the comparison of the skin elasticity change rate of examples 1 to 5 of application and comparative examples 1 to 8 of the present invention.

Detailed Description

Various exemplary embodiments, features and aspects of the invention will be described in detail below. The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration. Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.

Furthermore, in the following detailed description, numerous specific details are set forth in order to provide a better understanding of the present invention. It will be understood by those skilled in the art that the present invention may be practiced without some of these specific details. In other instances, methods, means, devices and steps which are well known to those skilled in the art have not been described in detail so as not to obscure the invention.

It should be noted that:

in the present specification, the meaning of "may" or "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

In the present specification, the numerical range represented by "numerical value a to numerical value B" means a range including the end point numerical value A, B.

All units used in the present invention are international standard units unless otherwise stated, and numerical values and numerical ranges appearing in the present invention should be understood to include errors allowed in industrial production.

In the present specification, reference to "some particular/preferred embodiments," "other particular/preferred embodiments," "embodiments," and the like, means that a particular element (e.g., feature, property, and/or characteristic) described in connection with the embodiment is included in at least one embodiment described herein, and may or may not be present in other embodiments. In addition, it is to be understood that the described elements may be combined in any suitable manner in the various embodiments.

A first aspect of the invention provides a collagenase inhibitor comprising: the collagenase inhibitor comprises a pomegranate bark extract and a chamomile extract, wherein the addition amount of the pomegranate bark extract is 28-72% and the addition amount of the chamomile extract is 28-72% based on the total mass of the collagenase inhibitor.

The inventors of the present invention found that using a combination of a chamomile extract and a pomegranate rind extract, an excellent synergistic effect can be produced, and thus collagenase activity can be inhibited to achieve an anti-aging effect.

Specifically, in the present invention, the mass ratio of the pomegranate bark extract to the chamomile extract is 1: 0.39-2.5, preferably 1: 0.4-2.4, more preferably 1: 0.42-2.2, further preferably 1: 0.45-2.1, further preferably 1: 0.5-2, and further preferably 1: 0.55-1.8. When the mass ratio of the pomegranate rind extract to the chamomile extract is within the above range, a synergistic effect can be further exhibited, and the collagenase activity inhibition effect is excellent.

The method for preparing the collagenase inhibitor of the present invention is not particularly limited, and may be a method generally used in the art, and specifically may include a step of mixing the components of the collagenase inhibitor. For example, the pomegranate rind extract and the chamomile extract are mixed uniformly by conventional mixing means.

Collagenase

Collagenases belong to one class of Matrix Metalloproteinases (MMPs). Matrix metalloproteinases are a family of endopeptidases with a zinc ion-dependent biological activity and the ability to degrade the extracellular Matrix (ECM). Collagenase mainly acts to degrade collagen and elastin in dermis, and Tissue Type Inhibitor (TIMPs) thereof specifically inhibits the activity of collagenase by covalently binding with highly conserved zinc binding site, co-regulates the metabolism of matrix, and maintains the structure of dermis.

The collagenase includes interstitial collagenase (MMP-1), polymorphonuclear collagenase (MMP-8) and collagenase 3 (MMP-13). Among them, interstitial collagenase, also known as collagenase-1, has various functions and can act on different substrates. Interstitial collagenase degrades several matrix molecules, such as aggrecan, multipotent glycans, basement membrane proteoglycans, casein, nidogen, serine protein inhibitors, and mucin-C. Thus, interstitial collagenases play a key role in the physiological repair of the extracellular matrix. The invention is mainly based on the important function of interstitial collagenase in the skin aging process, and inhibits the activity of interstitial collagenase, thereby reducing the inflammatory reaction and wrinkles of the skin, and being used as a way for delaying aging to test the anti-aging function.

It is to be noted that the object of the collagenase inhibitor of the present invention is to inhibit collagenase activity, for example: inhibit the activity of interstitial collagenase, but not the expression of collagenase.

Pomegranate rind extract

Pomegranate (Punica grandium L.) is deciduous shrub or tree of the genus Punica (Punicaceae) of the family Punicaceae. Native balkan peninsula to iran and its neighborhood, both temperate and tropical areas of the world are cultivated. China has cultivation in the south and north, and the planting area is large in Jiangsu, Henan and other fields. Pomegranate is one kind of pomegranate.

The pomegranate rind contains abundant chemical substances, such as tannins, alkaloids, flavonoids, amino acids and organic acids, which have pharmacological and health promoting effects on human body, and can be used for treating anthelmintic, hemostatic, dysentery and toothache. The red pomegranate bark extract is rich in pomegranate bark polyphenol, which comprises ellagitannin, ellagic catechin, chlorogenic acid, etc. The substances have effects of resisting oxidation, scavenging free radicals, preventing sunburn, whitening skin, etc.

In the invention, the added amount of the pomegranate bark extract is 28-72% by mass of the total mass of the collagenase inhibitor. When the addition amount of the pomegranate bark extract is 28-72%, collagenase activity can be effectively inhibited.

Chamomile extract

Chamomile (Matricaria recutita L), also known as chamomile, is an annual herb of the chamomilla genus (Matricaria) of the family Compositae (Compositae). Chamomile is an important medicinal plant and spice raw material, and is mostly used as a medicine by using inflorescence or whole herbs. Chamomile is sweet, fragrant, slightly bitter in taste and rich in bioactive substances.

Essential oil extracted from flos Matricariae Chamomillae contains terpenoids, bisabolol, chamazulene, etc. as main ingredients, and has effects of diminishing inflammation, resisting allergy, relieving spasm, inhibiting bacteria, clearing heat, treating ulcer, etc.; the chamomile is rich in phenolic acid, coumarins and flavonoids, and has the effects of resisting oxidation, protecting liver, resisting vascular proliferation, diminishing inflammation, resisting allergy, resisting virus and the like.

In the invention, the chamomile extract is added in an amount of 28-72% by mass based on the total mass of the collagenase inhibitor. When the addition amount of the chamomile extract is 28-72%, collagenase activity can be effectively inhibited.

A second aspect of the invention provides a cosmetic liquid comprising a collagenase inhibitor according to the first aspect of the invention and a penetration enhancer; the cosmetic liquid can inhibit the activity of collagenase, particularly the activity of interstitial collagenase by adding a proper amount of collagenase inhibitor, so that the cosmetic liquid has excellent anti-aging effect and can improve the skin elasticity. In order to promote the absorption of collagenase inhibitors by the skin, the cosmetic liquid of the invention also uses penetration enhancers. By using a penetration enhancer, the collagenase inhibitor of the present invention can be exerted to a greater extent.

Wherein, the addition amount of the collagenase inhibitor is 0.01-10% of the total mass of the cosmetic liquid, such as: 0.5%, 1%, 2%, 3%, 5%, 6%, 7%, 8%, etc. When the collagenase inhibitor is added in an amount of 0.01-10%, the elasticity of the skin after the collagenase inhibitor is used is increased. When the addition amount of the collagenase inhibitor is less than 0.01%, the collagenase inhibitor cannot play a role in resisting aging; when the addition amount of the collagenase inhibitor is more than 10%, the content of the collagenase inhibitor is too high, the cost is too high, and the corresponding anti-aging effect is not obviously improved.

The addition amount of the penetration enhancer is 0.01-10% of the total mass of the cosmetic liquid. When the addition amount of the penetration enhancer is less than 0.01%, the penetration enhancing effect is not significant. When the addition amount of the penetration enhancer is more than 10%, the penetration enhancer cannot further function.

In the present invention, the penetration enhancer includes one or a combination of two or more of propylene glycol, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate, and chitosan. The invention preferably uses the combination of propylene glycol and bis-diethoxydiol cyclohexane 1, 4-dicarboxylate, and the propylene glycol and bis-diethoxydiol cyclohexane 1, 4-dicarboxylate have synergistic effect, so that the absorption effect of collagenase inhibitor can be more excellent.

When a combination of propylene glycol and bis-diethoxydiol cyclohexane 1, 4-dicarboxylate is used as a penetration enhancer, the amount of propylene glycol added is 0.01-5% and the amount of bis-diethoxydiol cyclohexane 1, 4-dicarboxylate added is 0.01-5% based on the total mass of the cosmetic liquid. When the addition amount of the propylene glycol is 0.01-5% and the addition amount of the bis-diethoxydiol cyclohexane 1, 4-dicarboxylate is 0.01-5%, the absorption effect of the collagenase inhibitor can be effectively improved. For example: the amount of propylene glycol added is 0.1%, 0.5%, 1%, 1.5%, 2.5%, 3.5%, 4.5%, etc., and the amount of bis-diethoxydiol cyclohexane 1, 4-dicarboxylate added is 0.1%, 0.5%, 1%, 1.5%, 2.5%, 3.5%, 4.5%.

The beauty lotion also comprises one or the combination of more than two of humectant, emulsifier, pH regulator, preservative, skin conditioner, skin whitening agent, allergy relieving agent, antioxidant, chelating agent and aromatic; the formula of the beauty fluid is mild, so that the efficacy of the collagenase inhibitor can be fully exerted. Specifically, the method comprises the following steps:

the addition amount of the humectant is 0.01-20% by the total mass of the cosmetic liquid. When the addition amount of the humectant is 0.01-20%, the humectant can play a role in keeping moisture. In order to further exert the efficacy of the humectant, the humectant of the present invention may be added in an amount of 1 to 18%, 3 to 16%, 5 to 14%, 6 to 12%, for example: 2%, 3%, 6%, 8%, 10%, 12%, 15%, 17%, 19%, etc.

In the invention, the humectant comprises one or a combination of more than two of propylene glycol, butanediol, 1, 2-pentanediol, panthenol, dipropylene glycol, glycerol polymethacrylate, glycerol polyacrylate, sodium hyaluronate and trehalose.

The addition amount of the emulsifier is 0.01-0.5% of the total mass of the cosmetic liquid. For example: may be 0.05-0.4%, may be 0.1-0.3%, etc. When the dosage of the emulsifier is less than 0.01%, the emulsification is insufficient, so that the system is unstable; when the dosage of the emulsifier is more than 0.5 percent, certain influence can be caused on the stability of the product.

The emulsifier comprises one or more of glycerol alcohol ether-25 PCA isostearate, PEG-10 campesterol, isosteareth-20, sorbitan sesquioleate, laureth-7, polyglycerol-3 methyl glucose distearate, PEG/PPG-10/1 polydimethylsiloxane and polyglycerol-3 diisostearate.

The invention uses the glycerol ether-25 PCA isostearate as an emulsifier, can moisten the skin, can form an oily protective film on the surface of the skin, reduces the damage of the outside to the skin and ensures that the skin is white and tender.

Since the emulsion system of the present invention is a non-water-in-oil system, when propylene glycol and bis-diethoxydiethylene glycol cyclohexane 1, 4-dicarboxylate are used as permeation promoters, their synergistic effect is not affected even if they are not added at the same time.

The addition amount of the skin conditioning agent is 0.01-5% of the total mass of the beauty liquid. Specifically, the skin conditioner may be added in an amount of 0.1 to 4%, may be 0.3 to 3%, may be 0.5 to 2%, or the like. When the addition amount of the skin conditioner is less than 0.01%, the corresponding effect cannot be achieved; when the skin conditioning agent is added in an amount of more than 5%, the cost is too high.

The skin conditioner comprises one or more of allantoin, Ulva lactuca extract, Phaeodactylum tricornutum extract, Macrocystis japonica extract, chlorella fermentation product, hydrolyzed collagen, brown algae extract, Viscum album leaf extract, Botanil Spirulina extract, lalang grass rhizome extract, and radix et caulis Opuntiae Dillenii extract.

The addition amount of the soothing and sensitizing agent is 0.01-5% of the total mass of the cosmetic liquid. Specifically, the amount of the sensitizer added may be 0.1 to 4%, 0.5 to 3%, 0.6 to 2%, or the like. The soothing agent can be properly added according to the needs. When the addition amount of the soothing and sensitizing agent is less than 0.01%, the corresponding effect cannot be achieved; when the addition amount of the sensitizer is more than 5%, the cost is too high.

The beauty lotion of the invention can also be added with some skin whitening agents. The skin whitening agent is added to play a role in brightening the skin color and achieve a certain whitening effect. The addition amount of the skin whitening agent is 0.01-5% of the total mass of the skin toner; for example, it may be 0.5%, 1%, 2%, 3%, 4%, etc. When the addition amount of the skin whitening agent is less than 0.01%, the content is too low to play a corresponding effect; when the skin conditioning agent is added in an amount of more than 5%, the cost is too high.

Specifically, the skin whitening agent comprises one or more of niacinamide, magnolia sieboldii extract, kojic acid and derivatives thereof, arbutin and derivatives thereof and vitamin C.

The addition amount of the pH regulator is 0.01-1% of the total mass of the cosmetic liquid. The pH value of the beauty fluid is more suitable for human skin by adding the pH regulator. Preferably, the amount of the pH adjuster of the present invention added may be 0.03 to 0.8%, 0.05 to 0.5%, 0.06 to 0.3%, or the like. When the amount of the pH regulator added is more than 1% or less than 0.01%, a cosmetic liquid having an appropriate pH value cannot be obtained. In the invention, the pH regulator comprises one or more of citric acid, sodium lactate, sodium citrate, sodium hydroxide, potassium hydroxide, triethanolamine, aminomethyl propanol and arginine.

The cosmetic liquid of the present invention may be appropriately added with an antioxidant and a chelating agent. Generally, the addition amount of the antioxidant is 0-2% of the total mass of the cosmetic liquid; the addition amount of the chelating agent is 0.01-1%. The antioxidant can be one or more of vitamin E, tocopherol acetate, butylated hydroxytoluene, lycopene, and ascorbic acid ethyl ether. The chelating agent may be disodium EDTA and/or tetrasodium EDTA and the like.

In addition, the beauty liquid of the invention also comprises a preservative and an aromatic. The addition amount of the preservative is 0.01-1.5% based on the total mass of the cosmetic liquid; the addition amount of the aromatic is 0.01-1%. The preservative can comprise one or more of phenoxyethanol, methyl hydroxybenzoate, propyl hydroxybenzoate, benzyl alcohol, benzoic acid and its salt. The aromatic may be a perfume, etc.

The beauty solution disclosed by the invention not only can play an anti-aging role, but also has a certain introduction effect, is easy to absorb by skin, enhances the effect of skin on skin care product absorption, and promotes the absorption of skin on subsequent maintenance. Various components in the formula can provide abundant nutrition for the skin, so that the skin is tender and smooth.

A third embodiment of the present invention provides a method for preparing the cosmetic liquid of the second embodiment, comprising a step of mixing the components of the cosmetic.

Specifically, taking the beauty fluid as an example, the preparation method of the beauty fluid comprises the following steps:

1. adding water, humectant, chelating agent, partial penetration enhancer, partial antiseptic, pH regulator, and partial skin conditioner into a stirring pot, stirring, and heating to 75-85 deg.C;

2. cooling to 40-50 deg.C, adding emulsifier, aromatic, residual penetration enhancer, collagenase inhibitor, residual skin conditioner, residual antiseptic, soothing agent, skin whitening agent, and optional antioxidant, and stirring;

3. cooling to 30-40 deg.C, discharging after qualified inspection, and standing for 12-48 hr;

4. and (5) after the inspection is qualified, subpackaging, packaging, inspecting again, and warehousing the finished product.

Examples

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

The red pomegranate peel extract was purchased from: quzhou City exhibition-macro biotechnology Co., Ltd;

chamomile extracts were purchased from: quzhou City exhibition-macro Biotechnology Ltd.

Comparative examples 1 to 5

Pomegranate rind extract is provided as collagenase inhibitor. Dissolving the red pomegranate peel extract in 5 groups of deionized water with different volumes to obtain 5 groups of test solutions, wherein the concentrations of the 5 groups of test solutions are 4000 mu g/mL, 3200 mu g/mL, 2400 mu g/mL, 1600 mu g/mL and 800 mu g/mL respectively. The log mass concentration of the pomegranate rind extract is shown in table 2 below and fig. 1.

Comparative examples 6 to 10

Chamomilla recutita extract is provided as a collagenase inhibitor. Chamomile extracts were dissolved in 5 groups of deionized water in volumes corresponding to comparative examples 1-5 to give 5 groups of test solutions, the concentrations of the 5 groups of test solutions being 4000. mu.g/mL, 3200. mu.g/mL, 2400. mu.g/mL, 1600. mu.g/mL, 800. mu.g/mL, respectively. The logarithmic mass concentration of the chamomile extract is shown in table 2 and figure 2 below.

Examples 1 to 5

Pomegranate rind extract and chamomile extract are provided as collagenase inhibitors. The collagenase inhibitor was obtained by mixing the pomegranate rind extract and the chamomile extract at a mass ratio of 3:1 (example 1), 2:1 (example 2), 1:1 (example 3), 1:2 (example 4), and 1:3 (example 5).

The collagenase inhibitor of example 1 was dissolved in 5 groups of deionized water to give 5 groups of test solutions, the concentrations of the 5 groups of test solutions were 2000. mu.g/mL, 1600. mu.g/mL, 1200. mu.g/mL, 800. mu.g/mL, 400. mu.g/mL, respectively.

The collagenase inhibitors of example 2 were dissolved in 5 groups of deionized water to give 5 groups of test solutions, the concentrations of the 5 groups of test solutions were 2000. mu.g/mL, 1600. mu.g/mL, 1200. mu.g/mL, 800. mu.g/mL, 400. mu.g/mL, respectively.

The collagenase inhibitors of example 3 were dissolved in 5 groups of deionized water to give 5 groups of test solutions, the concentrations of the 5 groups of test solutions were 2000. mu.g/mL, 1600. mu.g/mL, 1200. mu.g/mL, 800. mu.g/mL, 400. mu.g/mL, respectively.

The collagenase inhibitors of example 4 were dissolved in 5 groups of deionized water to give 5 groups of test solutions, the concentrations of the 5 groups of test solutions were 2000. mu.g/mL, 1600. mu.g/mL, 1200. mu.g/mL, 800. mu.g/mL, 400. mu.g/mL, respectively.

The collagenase inhibitors of example 5 were dissolved in 5 groups of deionized water to give 5 groups of test solutions, the concentrations of the 5 groups of test solutions were 2000. mu.g/mL, 1600. mu.g/mL, 1200. mu.g/mL, 800. mu.g/mL, 400. mu.g/mL, respectively.

Wherein, the content (mass%) of the pomegranate rind extract and the chamomile extract in the collagenase inhibitor is shown in the following table 1, and the logarithmic mass concentration of the collagenase inhibitor is shown in the following table 3.

TABLE 1

Collagenase activity inhibition assay

The forskolin phenol reagent can be reduced by phenolic compounds to be blue (a mixture of molybdenum blue and tungsten blue) under alkaline conditions, and because the protein molecules contain amino acid containing phenolic groups (such as tyrosine, tryptophan and the like), the protein and the hydrolysate thereof can be subjected to the reaction, so that the protease activity can be measured by utilizing the principle. Generally, casein is used as a substrate, the casein is hydrolyzed by collagenase under certain pH value and temperature conditions, the enzymolysis reaction is stopped by trichloroacetic acid after a period of time, the supernatant is taken after the casein precipitate is removed by centrifugation or filtration, and Na is used₂CO₃Alkalizing, adding Folin phenol reagent, developing, wherein the shade of blue is proportional to the amount of tyrosine in the filtrate, and measuring with spectrophotometer at 650nm wavelength to calculate the activity of collagenase.

In the test, all collagenases used were matrix collagenase (MMP-1), purchased from: shanghai ze leaf Biotech Co., Ltd.

Collagenase activity is measured as the activity of collagenase that catalyzes the production of tyrosine by casein. The method specifically comprises the following steps:

taking 1 test tube, respectively adding 0.25mL of deionized water and 0.25mL (32U/mL) of collagenase, then adding 0.5mL (1.0%, w/v) of a substrate casein solution, immediately mixing the solutions, preserving the heat in a water bath at 37 ℃ for 10min, then adding 1mL (6.5%, w/v) of a trichloroacetic acid solution, mixing the solutions uniformly, standing the mixture for 10min, centrifuging the mixture for 5min at 10000rpm, taking 0.5mL of a supernatant sample in the test tube, respectively adding 0.5mL (mass concentration: 10%) of a sodium bicarbonate test solution into another test tube, and shaking the test tube uniformly. After 10 minutes, respectively adding 0.5ml of forinophenol test solution (diluted by 2 times by 1mol/L acid concentration of the forinophenol test solution), immediately mixing uniformly, and standing for 30 minutes at room temperature; the supernatant 1 of the experimental group was obtained and the absorbance at a wavelength of 650nm was measured and recorded as A1.

② taking another 1 test tube, respectively adding 0.25mL of deionized water and 0.25mL (32U/mL) of collagenase, then adding 1mL (6.5%, w/v) of trichloroacetic acid solution and immediately mixing, preserving heat in 37 ℃ water bath for 10min, then adding 0.5mL (1.0%, w/v) of substrate casein solution and mixing, standing for 10min, centrifuging at 10000rpm for 5min, taking 0.5mL of supernatant sample in the test tube, respectively adding 0.5mL (mass concentration: 10%) of sodium bicarbonate test solution into another test tube, and shaking uniformly. After 10 minutes, respectively adding 0.5ml of forinophenol test solution (diluted by 2 times by 1mol/L acid concentration of the forinophenol test solution), immediately mixing uniformly, and standing for 30 minutes at room temperature; control supernatant 2 was obtained and the absorbance measured at 650nm and recorded as A2.

③ respectively adding 0.25mL (32U/mL) of collagenase into 35 test tubes, then respectively adding 0.25mL (1.0%, w/v) of the test solution of the comparative examples 1-10 and the examples 1-5, and mixing uniformly, then adding 0.5mL (1.0%, w/v) of the casein solution of the substrate and immediately mixing uniformly, after preserving heat in a water bath at 37 ℃ for 10min, then adding 1mL (6.5%, w/v) of trichloroacetic acid solution and mixing uniformly, standing for 10min, centrifuging at 10000rpm for 5min, respectively taking 0.5mL of the supernatant sample in 35 test tubes, respectively adding 0.5mL of sodium bicarbonate test solution into another 35 test tubes, respectively adding 0.5mL (mass concentration: 10%) of sodium bicarbonate test solution, and shaking uniformly. After 10 minutes, respectively adding 0.5ml of forinophenol test solution (diluted by 2 times by 1mol/L acid concentration of the forinophenol test solution), immediately mixing uniformly, and standing for 30 minutes at room temperature; 35 groups of experimental group supernatants 3 were obtained and absorbance was measured at 650nm and recorded as A3.

And fourthly, respectively adding 0.25mL (32U/mL) of collagenase into 35 test tubes, then respectively adding 0.25mL of the test solution of the comparative examples 1-10 and the examples 1-5, uniformly mixing, then adding 1mL (6.5%, w/v) of trichloroacetic acid solution, immediately uniformly mixing, keeping the temperature in a water bath at 37 ℃ for 10min, then adding 0.5mL (1.0%, w/v) of casein solution as a substrate, uniformly mixing, standing for 10min, centrifuging at 10000rpm for 5min, respectively taking 0.5mL of supernatant samples in the 35 test tubes, respectively adding 0.5mL (mass concentration: 10%) of sodium bicarbonate test solution into the other 35 test tubes, and uniformly shaking. After 10 minutes, respectively adding 0.5ml of forinophenol test solution (diluted by 2 times by 1mol/L acid concentration of the forinophenol test solution), immediately mixing uniformly, and standing for 30 minutes at room temperature; 35 control group supernatant 4 was obtained and absorbance was measured at 650nm and recorded as A4.

The inhibition rate is [1- (A3-A4)/(A1-A2) ] x 100%

In the formula: a1 is the absorbance of supernatant 1 of experimental group without adding collagenase inhibitor;

a2 is the absorbance of control supernatant 2 without collagenase inhibitor;

a3 is the absorbance of supernatant 3 of experimental group containing collagenase inhibitor;

a4 is the absorbance of control supernatant 4 containing collagenase inhibitor.

The respective collagenase activity inhibition rates of the pomegranate rind extract (comparative examples 1 to 5) and the chamomile extract (comparative examples 6 to 10) were calculated. Combining the logarithmic mass concentration-collagenase activity inhibition ratio relationship chart (figure 1 and figure 2), and calculating the corresponding mass concentration (IC) when the inhibition ratio of the pomegranate bark extract is 50%_50A) Mass concentration (IC) corresponding to 50% inhibition ratio of chamomile extract_50B) The results are shown in Table 2.

TABLE 2

Then, the collagenase activity inhibition rates of the collagenase inhibitors of examples 1 to 5 were measured. And combining the logarithmic mass concentration-collagenase activity inhibition ratio relation diagram (figure 3), and calculating the mass concentration (IC) of the pomegranate bark extract when the pomegranate bark extract and the chamomile extract have the composite effect to generate the equivalent inhibition ratio (50 percent) by conversion_50a) Mass concentration of chamomile extract ((IC) when the combined effect of pomegranate rind extract and chamomile extract produces an equivalent inhibition rate (50%) ((IC)_50b) The results are shown in Table 3.

The effect of the combined effect of the pomegranate rind extract and the chamomile extract can be evaluated by the interaction coefficient γ, and the specific results are shown in table 3.

γ＝IC_50a/IC_50A+IC_50b/IC_50B

Wherein, IC_50ARepresents the corresponding mass concentration when the inhibition rate of the pomegranate bark extract is 50%;

IC_50Bshows the inhibition of chamomile extractThe mass concentration corresponding to the preparation rate of 50 percent;

IC_50athe mass concentration of the pomegranate bark extract is shown when the compound action of the pomegranate bark extract and the chamomile extract generates an equivalent inhibition rate (50%);

IC_50bthe mass concentration of the chamomile extract is shown when the combined action of the pomegranate rind extract and the chamomile extract generates an equivalent inhibition rate (50%).

Wherein γ ═ 1, indicates that the pomegranate rind extract and chamomile extract exhibit a simple additive effect; gamma is less than 1, the pomegranate rind extract and the chamomile extract show a synergistic effect, and the smaller the gamma value is, the stronger the synergistic effect is; gamma is more than 1, the pomegranate rind extract and the chamomile extract show antagonistic effect, and the larger the gamma value is, the larger the antagonistic effect is.

TABLE 3

Note: in Table 3, examples 1 and 5 are embodied in the present invention as reference examples 1 and 5, which can be compared with examples 2 to 4.

As can be seen from table 3, the collagenase inhibitor of the present invention has an interaction coefficient of less than 1, and the interaction coefficient value thereof may be 0.8 or less, and even 0.7 or less, so that the pomegranate rind extract and the chamomile extract may exhibit an excellent synergistic effect.

Application examples 1 to 5

The cosmetic liquids were prepared according to the contents (mass percentages) of the components in the cosmetic liquid formulations of application examples 1 to 5 in table 4 below, and according to the following production process steps. The production process comprises the following steps:

1. adding the A, B phase raw materials into a stirring pot, stirring and heating to 83 ℃;

2. cooling to 42 ℃, adding C, D phase and stirring evenly;

3. cooling to 37 ℃, discharging after the inspection is qualified, and standing for 24 hours;

Note: the A, B, C, D phases in the process are respectively

Phase A: water;

phase B: butanediol, 1, 2-pentanediol, propylene glycol, dipropylene glycol, sodium hyaluronate, allantoin, glycerol, panthenol, citric acid, trehalose, sodium citrate, disodium EDTA, glycerol polymethacrylate, and methylparaben;

and C phase: glycol ether-25 PCA isostearate, essence, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate;

phase D: pomegranate rind extract, chamomile extract, dipotassium glycyrrhizinate, Bolinate spirulina extract, nicotinamide and benzyl alcohol.

Wherein butanediol, 1, 2-pentanediol, dipropylene glycol, sodium hyaluronate, glycerol, panthenol, trehalose and glycerol polymethacrylate are used as humectants;

citric acid and sodium citrate are pH regulators; EDTA disodium is a chelating agent;

propylene glycol and bis-diethoxydiol cyclohexane 1, 4-dicarboxylic acid ester are taken as penetration enhancers;

glycol ether-25 PCA isostearate as emulsifier; dipotassium glycyrrhizinate is used as a allergy relieving agent;

the pomegranate rind extract and the chamomile extract are collagenase inhibitors;

the Bonnate Spirulina extract and allantoin are skin conditioner; niacinamide is a skin whitening agent;

methyl hydroxybenzoate and benzyl alcohol as antiseptic; the essence is aromatic.

TABLE 4

Application of comparative examples 1 to 8

The cosmetic liquids were prepared according to the contents (mass percentages) of the components in the cosmetic liquid formulations of comparative application examples 1 to 8 in the following table 5, and in the same manner as in application examples 1 to 5.

TABLE 5

Skin elasticity test

Method for testing skin elasticity: the test principle is based on the principle of suction and stretching, where a negative pressure is generated on the skin surface to be tested to suck the skin into a specific test probe, and the depth of the skin sucked into the test probe is measured by a non-contact optical test system. The test probe includes a transmitter and receiver of light, the ratio of which (the ratio of transmitted light to received light) is proportional to the depth of skin being absorbed, thus obtaining a time-dependent curve of the length of skin stretched.

Measuring the skin elasticity of the subject by using a probe PVM600 and a skin elasticity measuring instrument MPA580 of German CK company, selecting a parameter R2 as a comparison index (R2: the ratio of the skin rebound quantity without negative pressure to the maximum stretching quantity with negative pressure is closer to 1, the skin elasticity is better) and measuring for 3 times in total, and taking an average value; the improvement of the skin elasticity of the tested area by the product was evaluated by measuring the change in the skin elasticity value R2 before and after use of the product.

The number of the subjects was 33, the test period was 8 weeks, the test selected the cosmetic liquids of application examples 1 to 5 and the cosmetic liquids of application comparative examples 1 to 8 as test samples, the subjects were divided into 13 different areas on the forearm, and the cosmetic liquids of application examples 1 to 5 and the cosmetic liquids of application comparative examples 1 to 8 were applied to the different areas on the inner side of the forearm every morning and evening, and the application amount was about 2mg/cm²And the position of application of each test sample was kept constant during the test period, and then the skin elasticity of the test area before the test and at 8 weeks of use was measured, respectively, to further characterize the rate of change in skin elasticity, and the results of the specific rate of change in elasticity (averaged) are shown in table 6 and fig. 5.

TABLE 6

As can be seen from table 6 and fig. 5, the application examples 1 to 5 of the present application have a large change rate of elasticity, i.e., increased skin elasticity, and thus, the use of the pomegranate rind extract and the chamomile extract is effective in improving skin aging.

In the application comparative examples 1 to 8 of the present application, the change rate of skin elasticity is small, and thus, the anti-aging effect is relatively poor in the application comparative examples 1 to 8.

The above examples of the present invention are merely examples for clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A cosmetic liquid comprising a collagenase inhibitor and a penetration enhancer; the addition amount of the collagenase inhibitor is 0.01-10% of the total mass of the cosmetic liquid; the addition amount of the penetration enhancer is 0.01-10%;

2. The cosmetic liquid according to claim 1, wherein the mass ratio of the pomegranate rind extract to the chamomile extract is 1: 0.39-2.5, preferably 1: 0.4-2.4, more preferably 1: 0.42-2.2, further preferably 1: 0.45-2.1, further preferably 1: 0.5-2, and further preferably 1: 0.55-1.8.

3. The cosmetic liquid according to claim 1 or 2, wherein the penetration enhancer comprises one or a combination of two or more of propylene glycol, bis-diethoxydiethylene glycol cyclohexane 1, 4-dicarboxylate and chitosan.

4. The cosmetic liquid according to any one of claims 1 to 3, wherein the collagenase is interstitial collagenase.

5. The cosmetic liquid according to any one of claims 1 to 4, further comprising one or a combination of two or more of a moisturizing agent, an emulsifying agent, a pH adjusting agent, a preservative, a skin conditioning agent, a skin whitening agent, a soothing agent, an antioxidant, a chelating agent, and a fragrance;

6. The cosmetic liquid according to claim 5, wherein the skin conditioner comprises one or a combination of two or more of allantoin, Ulva lactuca extract, Phaeodactylum tricornutum extract, Macrocystis japonica extract, Chlorella fermentation product, hydrolyzed collagen, Brown algae extract, Viscum album Biloba leaf extract, Botanil Spirulina extract, lalang grass rhizome extract, and Opuntia ficus extract.

7. The cosmetic liquid according to claim 5 or 6, wherein the skin whitening agent comprises one or a combination of two or more of niacinamide, magnolia sieboldii extract, kojic acid and its derivatives, arbutin and its derivatives, vitamin C; and/or the presence of a gas in the gas,

8. A collagenase inhibitor comprising: the collagenase inhibitor comprises a pomegranate bark extract and a chamomile extract, wherein the addition amount of the pomegranate bark extract is 28-72% and the addition amount of the chamomile extract is 28-72% based on the total mass of the collagenase inhibitor.

9. The collagenase inhibitor according to claim 8, wherein the mass ratio of the pomegranate rind extract to the chamomile extract is 1: 0.39-2.5, preferably 1: 0.4-2.4, more preferably 1: 0.42-2.2, further preferably 1: 0.45-2.1, further preferably 1: 0.5-2, and further preferably 1: 0.55-1.8.

10. A method of preparing a collagenase inhibitor according to any one of claims 8 or 9, comprising the step of mixing the components of the collagenase inhibitor.