CN113041177A

CN113041177A - Collagenase inhibitor, eye essence containing the same and preparation method thereof

Info

Publication number: CN113041177A
Application number: CN201911377548.7A
Authority: CN
Inventors: 杨登亮; 谢佩佩; 李传茂; 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
Anticipated expiration: 2039-12-27
Also published as: CN113041177B

Abstract

The invention provides a collagenase inhibitor, eye essence containing the collagenase inhibitor and a preparation method of the eye essence. The collagenase inhibitor comprises a rhodiola crenulata extract and a chamomile extract, and the addition amount of the rhodiola crenulata extract is 3-30% and the addition amount of the chamomile extract is 70-97% based on the total mass of the collagenase inhibitor. The collagenase inhibitor of the present invention has an excellent inhibitory effect on collagenase activity and has no side effects on the human body.

Description

Collagenase inhibitor, eye essence containing the same and preparation method thereof

Technical Field

The invention relates to a collagenase inhibitor, eye essence containing the collagenase inhibitor and a preparation method of the eye essence, and belongs 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 firstly provides a collagenase inhibitor and a preparation method thereof, and the polypeptide and the retinol have certain irritation and safety risks to skin.

Further, the present invention also provides an eye essence comprising the collagenase inhibitor, which has an excellent anti-aging effect.

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

Means for solving the problems

The present invention provides a collagenase inhibitor comprising: the collagenase inhibitor comprises a rhodiola crenulata extract and a chamomile extract, wherein the adding amount of the rhodiola crenulata extract is 3-30% and the adding amount of the chamomile extract is 70-97% based on the total mass of the collagenase inhibitor.

The collagenase inhibitor comprises, by mass, 1: 4-32, preferably 1: 5-30, more preferably 1: 8-28, even more preferably 1: 10-27, even more preferably 1: 12-26, and even more preferably 1: 15-25 of rhodiola crenulata extract and chamomile extract.

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

The present invention also provides a method for preparing a collagenase inhibitor according to the present invention, comprising the step of mixing the components of a collagenase inhibitor.

The invention also provides an eye essence which comprises the collagenase inhibitor and the penetration enhancer, wherein the addition amount of the collagenase inhibitor is 0.01-10% of the total mass of the eye essence; the addition amount of the penetration enhancer is 0.01-10%.

The eye essence comprises one or more of propylene glycol, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate and chitosan.

The eye essence comprises one or more of a humectant, a thickener, a pH regulator, a preservative, an emulsifier, grease, a skin conditioner, a skin whitening agent, a soothing agent and an aromatic;

preferably, the addition amount of the humectant is 0.01-20% by mass of the total eye essence; the addition amount of the thickening agent is 0.02-0.8%; 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 emulsifier is 0.01-2%; the addition amount of the grease is 1-8%; 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 aromatic is 0.005-0.5%.

The eye essence comprises one or more of a giant kelp extract, an oat peptide, ceramide 2, an fucus extract, a chlorella fermentation product, hydrolyzed collagen, hydrolyzed wheat protein, a green algae extract, a brown algae extract, allantoin, a lactobacillus/soybean fermentation product extract, a ginkgo mistletoe leaf extract, a cogongrass rhizome extract, a cactus extract, trehalose and a clerodendranthus spicatus extract.

The eye essence 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 Hamamelis virginiana flower water, centella asiatica extract, ginger root extract, bisabolol, and dipotassium glycyrrhizinate.

The present invention also provides a method for preparing the eye essence according to the present invention, which comprises the step of mixing the components of the eye essence.

ADVANTAGEOUS EFFECTS OF INVENTION

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 eye essence disclosed by the invention is mild in formula, and can effectively improve the skin elasticity, so that the anti-aging effect is achieved.

The preparation method of the eye essence is simple to operate, easily available in raw materials and capable of realizing batch production.

Drawings

FIG. 1 is a graph showing the logarithmic mass concentration-collagenase activity inhibition ratios of rhodiola crenulata extracts of 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 rhodiola crenulata extract in collagenase inhibitors according to examples 1 to 5 of the present invention and the interaction coefficient.

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 rhodiola crenulata extract and a chamomile extract, wherein the adding amount of the rhodiola crenulata extract is 3-30% and the adding amount of the chamomile extract is 70-97% 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 rhodiola crenulata extract, an excellent synergistic effect can be produced, and thus the activity of collagenase can be inhibited to achieve an anti-aging effect.

Specifically, in the invention, the mass ratio of the rhodiola crenulata extract to the chamomile extract is 1: 4-32, preferably 1: 5-30, more preferably 1: 8-28, further preferably 1: 10-27, further preferably 1: 12-26, and further preferably 1: 15-25. When the mass ratio of the rhodiola crenulata 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 rhodiola crenulata extract and the chamomile extract are mixed uniformly by a conventional mixing method.

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 its Tissue Type Inhibitor (TIMPs) specifically inhibits the activity of collagenase by covalently binding with its highly conserved zinc binding site, co-regulates the metabolism of the matrix, and maintains the structure of the 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 realize 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.

Rhodiola crenulata extract

Rhodiola crenulata (hook.f. et Thoms.) H.Ohba) is a perennial herb of Rhodiola (Rhodiola) of Crassulaceae (Crassulaceae). Rhodiola crenulata is produced mostly in Tibet, northwest of Yunnan and West of Sichuan, and generally in hillside grassland, shrubs and stone cracks with elevation of 2800-.

Rhodiola crenulata belongs to a common traditional Chinese medicine plant, has the effects of promoting blood circulation, stopping bleeding, resisting radiation, resisting fatigue, enhancing the immunocompetence of a human body and the like, and has very important clinical effects. The chemical components of rhodiola crenulata mainly comprise alcohol glycosides, flavones, polysaccharides, volatile oils and the like, and the total flavones in the rhodiola plants have various pharmacological activities of oxidation resistance, virus resistance, tumor resistance and the like.

In the invention, the rhodiola crenulata extract is added in an amount of 3-30% by mass based on the total mass of the collagenase inhibitor. When the addition amount of the rhodiola crenulata extract is 3-30%, the collagenase activity can be effectively inhibited. Specifically, the rhodiola crenulata extract may be added in an amount of 5%, 8%, 10%, 12%, 15%, 18%, 20%, 22%, 25%, 28%, etc.

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 70-97% by mass based on the total mass of the collagenase inhibitor. When the addition amount of the chamomile extract is 70-97%, the collagenase activity can be effectively inhibited. Specifically, the added amount of the chamomile extract may be 72%, 75%, 78%, 80%, 82%, 85%, 88%, 90%, 92%, 95%, or the like.

A second aspect of the invention provides an ophthalmic concentrate comprising a collagenase inhibitor according to the first aspect of the invention and a penetration enhancer; the eye essence disclosed by the invention can inhibit the activity of collagenase, particularly the activity of interstitial collagenase by adding a proper amount of collagenase inhibitor, so that the eye essence disclosed by the invention has an excellent anti-aging effect and can improve the skin elasticity. In order to promote the absorption of collagenase inhibitors into the skin, the eye essence of the present 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% based on the total mass of the eye essence, 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 penetration enhancer is added in an amount of 0.01-10% by mass based on the total mass of the eye essence. 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 eye essence. 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 eye essence also comprises one or the combination of more than two of a humectant, a thickening agent, a pH regulator, a preservative, an emulsifier, grease, a skin conditioner, a skin whitening agent, a soothing agent and an aromatic. The eye essence disclosed by the invention is mild in formula composition, and can fully exert the efficacy of the collagenase inhibitor disclosed by the invention. Specifically, the method comprises the following steps:

the addition amount of the humectant is 0.01-20% by total mass of the eye essence. When the addition amount of the humectant is 0.01-20%, the humectant can play a role in moisturizing and hydrating. In order to further exert the efficacy of the moisturizer, the amount of the moisturizer of the present invention added may be preferably 1 to 19%, 4 to 18%, 6 to 17%, 8 to 16%, 10 to 15%, or the like. When the content of the humectant is less than 0.01%, the moisturizing effect is not obvious; when the content of the moisturizer is more than 20%, the eye essence is sticky.

In the present invention, the moisturizer comprises one or a combination of two or more of PEG/PPG-17/6 copolymer, dipropylene glycol, butylene glycol, panthenol, glycerin, polyethylene glycol-32, glyceryl polyether-26, sodium hyaluronate, polyacrylic acid, and a complex of betaine and glycerin. The eye essence of the present invention can have excellent moisturizing properties using a combination of a plurality of moisturizers.

According to the invention, by using the compound of polyacrylic acid, betaine and glycerol, more excellent moisturizing effect can be obtained.

The addition amount of the thickening agent is 0.02-0.8% of the total mass of the eye essence. When the addition amount of the thickener is between 0.02 and 0.8 percent, the low-viscosity feeling and excellent use feeling are achieved, the dispersibility is good, and the absorption is fast. When the addition amount of the thickener is less than 0.02%, the eye essence is thin in texture and does not have any sticky feeling; when the thickener is added in an amount of more than 0.8%, the eye essence is too thick and heavy, which may increase the burden on the skin.

In the present invention, the thickener includes one or more of acrylic acid (ester)/C10-30 alkanol acrylate crosspolymer, hydroxyethyl cellulose, xanthan gum, hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, carbomer, isohexadecane and polysorbate-60, a complex of hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and sorbitan isostearate, and ammonium acryloyldimethyl taurate/VP copolymer.

The invention uses the compound thickener such as isohexadecane, polysorbate-60, hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, sorbitan isostearate and the like, and can improve the stability of the eye essence.

The addition amount of the pH regulator is 0.01-1% of the total mass of the eye essence. The pH value of the eye essence 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%, 0.06 to 0.5%, 0.1 to 0.3%, or the like. When the amount of the pH regulator added is more than 1% or less than 0.01%, no eye essence with a proper pH value can be obtained. The pH regulator comprises one or more of aminomethyl propanol, arginine, citric acid, sodium citrate and sodium hydroxide.

The addition amount of the emulsifier is 0.01-2% by the total mass of the eye essence, such as: may be 0.1 to 1.5%, may be 0.5 to 1.2%, 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 2 percent, certain influence can be caused on the stability of the product.

In the present invention, the emulsifier includes one or a combination of two or more of polysorbate-20, PEG-20 methylglucose sesquistearate, sorbitan isostearate, polyglyceryl-3 methylglucose distearate, laureth-7, isosteareth-20.

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.

In the invention, the addition amount of the oil is 1-8% by total mass of the eye essence. The amount of the oil or fat added may be 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, etc. By adding oil into eye essence, evaporation of water on skin surface can be reduced, and skin chap can be prevented. In addition, by adding oil or fat, a hydrophobic film can be formed on the skin surface to prevent the invasion of harmful substances from the outside. When the content of the oil is less than 1%, evaporation of moisture on the skin surface cannot be reduced, and intrusion of harmful substances cannot be effectively prevented; when the content of the oil is more than 8%, the eye essence is too greasy, and the use feeling is reduced.

In the present invention, the oil or fat includes one or a combination of two or more of cyclopentadimethylsiloxane, polydimethylsiloxane, dimethiconol, oleyl erucate, shea butter, ethylhexyl palmitate, hydrogenated polydecene, cyclohexasiloxane, hydrogenated polyisobutene, C20-24 alkyl polydimethylsiloxane, C13-14 isoparaffin, and C12-15 alcohol benzoate.

In order to further improve the efficacy of the eye essence of the present application, the eye essence of the present invention further comprises a skin conditioner. The skin conditioner is added, so that the effects of moisturizing and moisturizing can be further achieved, and the generation of wrinkles can be reduced. In addition, the skin conditioner can also properly reduce the adhesive force of stratum corneum cells, accelerate the renewal of epidermal cells and enhance the repair capacity of skin.

The addition amount of the skin conditioner is 0.01-5% of the total mass of the eye essence; for example, it may be 0.5%, 1%, 2%, 3%, 4%, etc. When the addition amount of the skin conditioner 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 conditioner comprises one or more of a giant kelp extract, oat peptide, ceramide 2, a sea oak extract, a chlorella fermentation product, hydrolyzed collagen, hydrolyzed wheat protein, a green algae extract, a brown algae extract, allantoin, a lactobacillus/soybean fermentation product extract, a mistletoe ginkgo leaf extract, a cogongrass rhizome extract, a cactus extract, trehalose and a clerodendranthus spicatus extract.

The lactobacillus in the lactobacillus/fermented soybean product extract refers to a bacterium that can ferment saccharides to produce lactic acid. The lactobacillus/soybean fermentation product extract can be used as a skin conditioner in the invention, and has excellent moisturizing performance.

The clerodendranthus spicatus extract can be used as a skin conditioner, and can remarkably improve the oily appearance and pore size of skin and the uniformity and luster of the skin.

The eye essence can also be added with a small amount of skin whitening agent. 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% by the total mass of the eye essence; 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 amount of the skin whitening agent added is 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 eye essence of the invention can also be added with a small amount of soothing agents. By adding the allergy relieving agent, the skin can be calmed, so that the skin has certain allergy relieving effect on the injury red swelling of the facial skin, and the skin can be helped to resist inflammation, relieve and promote cell repair.

The addition amount of the allergy-relieving agent is 0.01-5% of the total mass of the eye essence; for example, it may be 0.5%, 1%, 2%, 3%, 4%, etc. When the addition amount of the allergy relieving agent is less than 0.01 percent, the allergy relieving effect is not obvious. When the addition amount of the allergy relieving agent is more than 5%, the further allergy relieving effect cannot be achieved, and the cost is too high.

The hamamelis virginiana flower water in the allergy-relieving agent belongs to high-concentration plant original dew, has light herbal fragrance, and has the effects of controlling oil, conditioning, astringing, tightening pores, removing stasis and relieving swelling. Can be used for caring skin and soothing skin.

In addition, the eye essence of the present invention may be appropriately added with an antioxidant and a chelating agent. Generally, the antioxidant is added in an amount of 0-2% by mass of the total eye essence; the addition amount of the chelating agent is 0-1%. The antioxidant can be one or more of vitamin E, tocopherol acetate, butylated hydroxytoluene, lycopene, etc. The chelating agent may be EDTA-2Na and/or EDTA-4Na, etc.

Finally, the preservative in the eye essence of the invention is added in an amount of 0.01-1.5%. The preservative can comprise one or the combination of more than two of phenoxyethanol, methyl hydroxybenzoate, propyl hydroxybenzoate, benzoic acid and salts thereof. In addition, the eye essence of the present invention further contains a fragrance. The addition amount of the aromatic is 0.005-0.5% by total mass of the eye essence. The aromatic may be a perfume, etc.

A third aspect of the present invention provides a method for preparing the eye essence of the second aspect, which comprises a step of mixing components of the eye essence.

Specifically, the preparation method comprises the following steps:

1. adding water, part of humectant, part of thickener and part of antiseptic into an emulsifying pot, stirring and heating to 80-85 deg.C;

2. slowly pumping part of the humectant and part of the penetration enhancer into an emulsifying pot, homogenizing for 1-5min, and stirring for 10-30min under heat preservation;

3. cooling to 62-68 deg.C, adding pH regulator, oil and fat and the rest thickener, homogenizing for 1-8min, and stirring;

4. cooling to 42-48 deg.C, adding the rest humectant, skin conditioner, skin soothing agent, skin whitening agent, the rest antiseptic, collagenase inhibitor, emulsifier, aromatic and the rest penetration enhancer, and stirring;

5. cooling to 35-40 deg.C, discharging, and standing for 12-48 hr;

6. 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.

Rhodiola crenulata extracts were purchased from: kunming Qiancao Biotechnology Co., Ltd;

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

Comparative examples 1 to 5

Rhodiola crenulata extract is provided as collagenase inhibitor. Dissolving the rhodiola crenulata 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 400 mu g/mL, 320 mu g/mL, 240 mu g/mL, 160 mu g/mL and 80 mu g/mL respectively. The log mass concentration of rhodiola crenulata 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

Rhodiola crenulata extract and chamomile extract are provided as collagenase inhibitors. The collagenase inhibitor was obtained by mixing rhodiola crenulata extract and chamomilla extract at a mass ratio of 1:1 (example 1), 1:10 (example 2), 1:20 (example 3), 1:25 (example 4), and 1:30 (example 5).

The collagenase inhibitors of example 1 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 400. mu.g/mL, 320. mu.g/mL, 240. mu.g/mL, 160. mu.g/mL, 80. 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 1000. mu.g/mL, 800. mu.g/mL, 600. mu.g/mL, 400. mu.g/mL, 200. 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 1000. mu.g/mL, 800. mu.g/mL, 600. mu.g/mL, 400. mu.g/mL, 200. 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 contents (mass%) of rhodiola crenulata extract and chamomile extract in the collagenase inhibitors are shown in the following table 1, and the logarithmic mass concentrations of the collagenase inhibitors are 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:

adding 0.25mL of deionized water and 0.25mL (32U/mL) of collagenase into 1 test tube respectively, then adding 0.5mL (1.0% w/v) of a substrate casein solution, immediately mixing the mixture, keeping the temperature in a water bath at 37 ℃ for 10min, then adding 1mL (6.5% w/v) of a trichloroacetic acid solution, mixing the mixture uniformly, standing the mixture for 10min, centrifuging the mixture for 5min at 10000rpm, taking 0.5mL of a supernatant sample in the test tube, adding 0.5mL (mass concentration: 10%) of a sodium bicarbonate test solution into another test tube respectively, 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 rhodiola crenulata 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 mass concentration (IC) corresponding to 50% inhibition ratio of rhodiola crenulata extract_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 rhodiola crenulata extract when the rhodiola crenulata extract and the chamomile extract have the equivalent inhibition ratio (50%) under the combined action_50a) Mass concentration of camomile extract ((IC) when rhodiola crenulata extract and camomile extract act in combination to produce an equivalent inhibition rate (50%))_50b) The results are shown in Table 3.

The effect of the combined effect of rhodiola crenulata extract and 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_50AThe mass concentration of the rhodiola crenulata extract is 50%;

IC_50Brepresents the mass concentration corresponding to the inhibition rate of the chamomile extract of 50%;

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

IC_50bthe mass concentration of the chamomile extract is shown when the rhodiola crenulata extract and the chamomile extract have equivalent inhibition rate (50%) due to the combined action.

Wherein, gamma is 1, which indicates that the rhodiola crenulata extract and the chamomile extract show simple additive effect; gamma is less than 1, the rhodiola crenulata 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 rhodiola crenulata 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, example 1 is embodied in the present invention as reference example 1, which can be compared with examples 2 to 5.

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 rhodiola crenulata extract and chamomile extract may exhibit excellent synergistic effects.

Application examples 1 to 5

The eye essence was prepared according to the contents (mass percentages) of the components in the eye essence formulations of application examples 1 to 5 in the following table 4, and according to the following production process steps. The production process comprises the following steps:

1. adding the phase A into an emulsifying pot, stirring and heating to 80-85 ℃;

2. slowly pumping the phase B into an emulsifying pot, homogenizing for 2min, and stirring for 15min under heat preservation;

3. cooling to 65 deg.C, adding C, D phase, homogenizing for 4min, and stirring;

4. cooling to 45 ℃, adding E, F, G phase and stirring evenly;

5. cooling to 37 ℃, discharging, and standing for 24 hours;

Note: the A, B, C, D, E, F, G phases in the process are respectively phase A: water, glyceryl polyether-26, PEG/PPG-17/6 copolymer, carbomer, and methylparaben;

phase B: dipropylene glycol, propylene glycol, glycerin, polyacrylic acid, and a complex of betaine and glycerin;

and C phase: aminomethyl propanol;

phase D: a complex of polydimethylsiloxane, isohexadecane and polysorbate-60 and hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and sorbitan isostearate;

phase E: butylene glycol, lactobacillus/soybean fermentation product extract, dipotassium glycyrrhizinate, hamamelis virginiana flower water, clerodendrum spicatum extract, and nicotinamide;

and (3) phase F: phenoxyethanol;

phase G: chamomile extract, rhodiola crenulata extract, polysorbate-20, essence and bis-diethoxydiol cyclohexane 1, 4-dicarboxylate.

The chamomile extract and the rhodiola crenulata extract used in the formula are collagenase inhibitors;

glycerin, dipropylene glycol, glyceryl polyether-26, PEG/PPG-17/6 copolymer, polyacrylic acid and betaine and glycerin complex, butylene glycol are humectants;

polydimethylsiloxane is a grease; niacinamide is a skin whitening agent;

polysorbate-20 is an emulsifier; aminomethyl propanol is a pH adjusting agent;

carbomer, isohexadecane and polysorbate-60 and a complex of hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and sorbitan isostearate are thickeners;

the lactobacillus/soybean fermentation product extract and the clerodendranthus spicatus extract are skin conditioners;

dipotassium glycyrrhizinate and Hamamelis virginiana flower water are soothing and sensitizing agents;

propylene glycol, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate are permeation enhancers;

phenoxyethanol and methyl hydroxybenzoate as antiseptic; the essence is an aromatic.

A complex of isohexadecane and polysorbate-60 and hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and sorbitan isostearate: the manufacturer: french Saybik Co.

TABLE 1

Application of comparative examples 1 to 8

The eye essence was prepared according to the contents (mass percentages) of the components in the eye essence formulations of comparative application examples 1 to 8 in the following table 5, and according to the same method 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 eye essences of application examples 1 to 5 and the eye essences of application comparative examples 1 to 8 as test samples, 13 different areas were divided on the forearm of the subject, the eye essences of application examples 1 to 5 and the eye essences of application comparative examples 1 to 8 were applied to 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 red pomegranate bark extract and the purslane 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 collagenase inhibitor comprising: the collagenase inhibitor comprises a rhodiola crenulata extract and a chamomile extract, wherein the adding amount of the rhodiola crenulata extract is 3-30% and the adding amount of the chamomile extract is 70-97% based on the total mass of the collagenase inhibitor.

2. The collagenase inhibitor according to claim 1, wherein the mass ratio of the rhodiola crenulata extract to the chamomile extract is 1: 4-32, preferably 1: 5-30, more preferably 1: 8-28, further preferably 1: 10-27, further preferably 1: 12-26, and further preferably 1: 15-25.

3. Collagenase inhibitor according to claim 1 or 2, characterised in that the collagenase is a interstitial collagenase.

4. A method of preparing a collagenase inhibitor according to any one of claims 1-3, comprising the step of mixing the components of a collagenase inhibitor.

5. An eye essence, which comprises the collagenase inhibitor and the penetration enhancer according to any one of claims 1 to 3, wherein the collagenase inhibitor is added in an amount of 0.01 to 10 percent by mass of the total mass of the eye essence; the addition amount of the penetration enhancer is 0.01-10%.

6. The eye essence of claim 5, wherein the penetration enhancer comprises one or more of propylene glycol, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate, and chitosan.

7. The eye essence according to claim 5 or 6, wherein the eye essence further comprises one or a combination of two or more of a humectant, a thickener, a pH regulator, a preservative, an emulsifier, an oil, a skin conditioner, a skin whitening agent, a soothing agent and a fragrance;

8. The eye essence according to claim 7, wherein the skin conditioner comprises one or a combination of two or more of a giant kelp extract, an avenin, ceramide 2, a fucus extract, a chlorella fermentation product, a hydrolyzed collagen, a hydrolyzed wheat protein, a green algae extract, a brown algae extract, allantoin, a lactobacillus/soybean fermentation product extract, a mistletoe ginkgo leaf extract, a cogongrass rhizome extract, a cactus extract, trehalose, and a clerodendranthus spicatus extract.

9. The eye essence according to claim 7 or 8, 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, and vitamin C; and/or the presence of a gas in the gas,

10. A method for preparing the eye essence according to any one of claims 5 to 9, comprising a step of mixing components of the eye essence.