CN113116762A

CN113116762A - Firming cream, preparation method thereof and collagenase inhibitor

Info

Publication number: CN113116762A
Application number: CN201911396221.4A
Authority: CN
Inventors: 杨登亮; 林盛杰; 李传茂; 邱桂芳; 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: 2019-12-30
Filing date: 2019-12-30
Publication date: 2021-07-16
Anticipated expiration: 2039-12-30
Also published as: CN113116762B

Abstract

The invention provides firming cream, a preparation method thereof and a collagenase inhibitor. The firming cream comprises a collagenase inhibitor and a penetration enhancer, wherein the adding amount of the collagenase inhibitor is 0.01-10% of the total mass of the firming cream; the addition amount of the penetration enhancer is 0.01-10%; the collagenase inhibitor comprises pomegranate rind extract and grapefruit extract; based on the total mass of the collagenase inhibitor, the addition amount of the pomegranate bark extract is 15-65%, and the addition amount of the grapefruit extract is 35-85%. The firming cream disclosed by the invention is mild in formula, and can effectively improve the skin elasticity, so that an anti-aging effect is achieved.

Description

Firming cream, preparation method thereof and collagenase inhibitor

Technical Field

The invention relates to firming cream, a preparation method thereof and a collagenase inhibitor, 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 provides a firming cream containing a collagenase inhibitor, which has excellent anti-aging performance.

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

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

Means for solving the problems

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

the collagenase inhibitor comprises pomegranate rind extract and grapefruit extract; based on the total mass of the collagenase inhibitor, the addition amount of the pomegranate bark extract is 15-65%, and the addition amount of the grapefruit extract is 35-85%.

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

The firming cream comprises the pomegranate rind extract and the grapefruit extract in a mass ratio of 1: 0.6-5.5, preferably 1: 0.8-5, more preferably 1: 1-4.5, further preferably 1: 1.2-4, further preferably 1: 1.5-3.5, and further preferably 1: 1.8-3.

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

The firming cream comprises one or more of a humectant, a thickener, a pH regulator, grease, a preservative, a skin conditioner, an emulsifier, a soothing agent, a chelating agent, an aromatic and an antioxidant; wherein the content of the first and second substances,

the addition amount of the humectant is 0.01-20%; the addition amount of the thickening agent is 0.02-5%; the addition amount of the pH regulator is 0.01-1%; the addition amount of the grease is 10-30%; 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 emulsifier is 0.01-5%; the addition amount of the allergy relieving agent is 0.01-5%; the addition amount of the antioxidant is 0.01-2%; the adding amount of the aromatic is 0.01-1%; the addition amount of the chelating agent is 0-2%.

The firming cream comprises one or more of manganese chloride, tapioca starch, ceramide 2, lactobacillus/soybean fermentation product extract, palmitoyl tripeptide-5, acetyl hexapeptide-8, palmitoyl oligopeptide, colloidal oat flour, hydrolyzed collagen, hydrolyzed wheat protein, kelp extract, allantoin, pansy extract and chlorella fermentation product.

The firming cream comprises Hamamelis virginiana water, stearyl glycyrrhetinate, bisabolol, ginger root extract and aloe extract, or the combination of more than two of the Hamamelis virginiana water, the stearyl glycyrrhetinate, the bisabolol, the ginger root extract and the aloe extract.

The invention also provides a preparation method of the firming cream, which comprises the step of mixing the components of the firming cream.

The present invention also provides a collagenase inhibitor comprising: the composition comprises a red pomegranate rind extract and a grapefruit extract, wherein the addition amount of the red pomegranate rind extract is 15-65% and the addition amount of the grapefruit extract is 35-85% based on the total mass of the collagenase inhibitor.

The collagenase inhibitor according to the present invention, wherein the mass ratio of the red pomegranate rind extract to the grapefruit extract is 1:0.6 to 5.5, preferably 1:0.8 to 5, more preferably 1:1 to 4.5, further preferably 1:1.2 to 4, further preferably 1:1.5 to 3.5, and further preferably 1:1.8 to 3.

ADVANTAGEOUS EFFECTS OF INVENTION

The firming cream disclosed by the invention is mild in formula, and can effectively improve the skin elasticity, so that an anti-aging effect is achieved.

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 firming cream is simple to operate, raw materials are easy to obtain, and the firming cream 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 grapefruit extracts of comparative examples 6 to 10 of the present 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 composition comprises a red pomegranate rind extract and a grapefruit extract, wherein the addition amount of the red pomegranate rind extract is 15-65% and the addition amount of the grapefruit extract is 35-85% based on the total mass of the collagenase inhibitor.

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

Specifically, in the invention, the mass ratio of the red pomegranate bark extract to the grapefruit extract is 1: 0.6-5.5, preferably 1: 0.8-5, more preferably 1: 1-4.5, further preferably 1: 1.2-4, further preferably 1: 1.5-3.5, and further preferably 1: 1.8-3. When the mass ratio of the red pomegranate rind extract to the grapefruit 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 commonly used in the art, for example, a method of mixing the red pomegranate rind extract and the grapefruit extract uniformly.

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 collagenase inhibitor of the present invention is intended to inhibit collagenase activity, not collagenase expression, for example: inhibiting the activity of interstitial 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 15-65% by the total mass of the collagenase inhibitor. When the addition amount of the pomegranate bark extract is 15-65%, the collagenase activity can be effectively inhibited.

Grapefruit extract

Grapefruit, also known as grapefruit (Citrus paradisi Macf), is a small tree of the genus Citrus (Citrus) of the family Rutaceae (Rutaceae). The grapefruit extract can nourish tissue cells, promote diuresis, reduce edema, resist bacteria and infection, treat large pores, and condition skin with mixed type, oily and the like which is easy to grow pox; deeply purifying oily skin sore and hyperemia skin, compacting skin heel tissue, and its component naringin has free radical chain reaction for blocking self-oxidation of pyrogallol, i.e. naringin has antioxidant effect.

In the present invention, the grapefruit extract is added in an amount of 35-85% by mass based on the total mass of the collagenase inhibitor. When the grapefruit extract is added in an amount of 35-85%, collagenase activity can be effectively inhibited.

A second aspect of the invention provides a firming cream comprising a collagenase inhibitor according to the first aspect of the invention and a penetration enhancer; according to the invention, the collagenase inhibitor with a proper content is added, so that the activity of collagenase, especially the activity of interstitial collagenase, can be inhibited, the firming cream disclosed by the invention has an excellent anti-aging effect, and the skin elasticity can be improved. In order to promote the absorption of collagenase inhibitors by the skin, the firming cream 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% of the total mass of the firming cream, 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 firming cream. 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 the 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 firming cream. 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 firming cream can also comprise one or more of a humectant, a thickening agent, a pH regulator, grease, a preservative, a skin conditioner, an emulsifier, a soothing agent, a chelating agent, an aromatic and an antioxidant. The formula of the firming cream 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% of the total mass of the firming cream. 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 is preferably 1 to 18%, 2 to 16%, 3 to 14%, 4 to 13%, 5 to 12%, 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 humectant is more than 20%, the firming cream has a sticky feeling.

In the invention, the humectant comprises one or a combination of more than two of dipropylene glycol, butanediol, glycerol, panthenol, glycerol polyacrylate, glycerol polyether-26, sodium hyaluronate, betaine and trehalose.

The addition amount of the grease is 10-30% of the total mass of the firming cream. For example: the addition amount of the grease can be 12%, 14%, 16%, 18%, 20%, 22%, 24%, 28% and the like. When the content of oil is 10-30%, it can not only form hydrophobic film on skin surface to prevent invasion of harmful substances, but also inhibit evaporation of water on skin surface to prevent skin chapping. When the amount of the added oil is less than 10%, invasion of harmful substances cannot be effectively prevented; when the addition amount of the oil is more than 30%, the firming cream 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 isopropyl myristate, polydimethylsiloxane, cyclomethicone, cyclopentadimethicone, caprylic/capric triglyceride, oleyl erucate, hydrogenated polyisobutene, shea butter, ethylhexyl palmitate, olive oil unsaponifiable matter, hydrogenated polydecene, cyclohexasiloxane, C20-24 alkyl dimethicone, C13-14 isoparaffin, and C12-15 alcohol benzoate.

The addition amount of the emulsifier is 0.01-5% of the total mass of the firming cream. Preferably 0.1 to 4%, for example: 0.5%, 1%, 1.2%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 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 5 percent, the irritation of the product can be increased, and meanwhile, certain influence can be caused on the stability of the product.

In the invention, the emulsifier comprises one or more of polyglycerol-3 methyl glucose distearate, PEG-100 stearate and glyceryl stearate complex, cetyl polyether-20, polysorbate-60, polyacrylamide and laureth-7 and C13-14 isoparaffin complex, and sorbitan sesquioleate.

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 thickening agent is 0.02-5% of the total mass of the firming cream. By adding the thickening agent, the firming cream of the invention can be made to have proper consistency and excellent stability. Preferably, the thickener of the present invention may be added in an amount of 0.05 to 4%, 0.1 to 3%, 1 to 3%, or the like.

The thickener comprises one or more of xanthan gum, behenyl alcohol, polyacrylamide, carbomer, polyethylene glycol-90M, acryloyl dimethyl taurate/VP copolymer, and hydroxyethyl acrylate/acryloyl dimethyl taurate copolymer.

In order to further improve the efficacy of the firming cream of the present application, the firming cream of the present invention further comprises the skin conditioning agent. The generation of wrinkles can be reduced by the addition of a skin conditioner. The effective components in the skin conditioner can penetrate into the deep part of skin and be absorbed by skin, thereby improving the state of skin.

The addition amount of the skin conditioning agent is 0.01-5% of the total mass of the firming cream. Preferably, the skin conditioning agent may be added in an amount of 0.1 to 4%, may be 0.5 to 3%, may be 0.6 to 2.5%, 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 conditioner is added in an amount of more than 5%, the cost is too high.

The skin conditioner comprises one or more of manganese chloride, cassava starch, ceramide 2, lactobacillus/soybean fermentation product extract, palmitoyl tripeptide-5, acetyl hexapeptide-8, palmitoyl oligopeptide, colloidal oat flour, hydrolyzed collagen, hydrolyzed wheat protein, giant kelp extract, allantoin, pansy extract and chlorella fermentation product.

The colloidal oat flour can reduce the irritation of the firming cream, relieve the itching symptom of the skin, and is used for diminishing inflammation, relieving itching and relieving erythema.

Among them, allantoin can reduce the adhesion of stratum corneum cells, accelerate epidermal cell renewal, enhance skin repair ability, and has high safety.

The manganese chloride can imitate in-vivo superoxide dismutase, can remove superoxide anions and other oxidation molecule precursors, and can delay skin aging and improve the health state of the skin when being used for the firming cream.

In order to further improve the efficacy of the firming cream, the firming cream further comprises a soothing agent. By adding the soothing agent, the skin can be calmed, so that the facial skin injury red swelling can be relieved. The addition amount of the soothing and sensitizing agent is 0.01-5% of the total mass of the firming cream. Specifically, the amount of the sensitizer added may be 0.1 to 4%, may be 0.2 to 3%, may be 0.3 to 2%, or the like. When the addition amount of the soothing and sensitizing agent is less than 0.01%, the corresponding effect cannot be achieved; when the amount of the sensitizer added is more than 5%, the cost is too high.

In the invention, the soothing agent comprises one or more of hamamelis water, stearyl alcohol glycyrrhetinate, bisabolol, ginger root extract and aloe extract.

The addition amount of the antioxidant is 0.01-2% of the total mass of the firming cream, such as: the antioxidant may be added in an amount of 0.1%, 0.3%, 0.5%, 0.8%, 1.0%, 1.2%, 1.5%, 1.8%, etc. The invention can remove free radicals, protect skin, reduce the damage of ultraviolet rays to skin, promote wound healing, prevent inflammation and prevent skin roughness and chap by adding a proper amount of antioxidant. The antioxidant can be one or more of tocopherol acetate, butylated hydroxytoluene, lycopene, etc.

The addition amount of the pH regulator is 0.01-1% of the total mass of the firming cream; the addition amount of the chelating agent is 0-1%. The pH regulator comprises one or more of aminomethyl propanol, arginine, citric acid, sodium citrate, and sodium hydroxide. The chelating agent may be EDTA-2Na and/or EDTA-4Na, etc.

In addition, the addition amount of the preservative in the firming cream is 0.01-1.5% based on the total mass of the firming cream; the addition amount of the aromatic is 0.01-1%. The preservative can comprise one or the combination of more than two of phenoxyethanol, methyl hydroxybenzoate, propyl hydroxybenzoate, benzoic acid and salts thereof. The aromatic may be a perfume, etc.

In a third aspect of the invention, there is provided a method of preparing the firming cream of the second aspect. The method comprises the step of mixing the components of the firming cream.

Specifically, the preparation method of the firming cream comprises the following steps:

1. adding oil, part of thickener, emulsifier, part of humectant, soothing agent, antioxidant, part of antiseptic, and aromatic into oil phase pan, stirring, and heating to 75-85 deg.C for dissolving completely;

2. adding part of humectant, part of penetration enhancer, optionally chelating agent, pH regulator, part of skin conditioner, and part of thickener into emulsifying pot, stirring, and heating to 75-85 deg.C for dissolving completely;

3. slowly pumping the oil phase substances in the oil phase pot into an emulsifying pot, stirring, homogenizing, vacuum emulsifying, and keeping the temperature of the emulsifying pot at 75-85 deg.C;

4. cooling to 40-50 deg.C, adding the rest humectant, the rest penetration enhancer, the rest thickener, collagenase inhibitor, the rest skin conditioner and the rest antiseptic, and stirring;

5. cooling to 30-40 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.

Red pomegranate rind extract: quzhou City exhibition-macro biotechnology Co., Ltd;

grapefruit extract: 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

Grapefruit extract is provided as a collagenase inhibitor. The grapefruit extract was dissolved in 5 sets of deionized water in volumes corresponding to comparative examples 1-5 to give 5 sets of test solutions, the concentrations of the 5 sets 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 grapefruit extract is shown in table 2 and fig. 2 below.

Examples 1 to 5

Pomegranate rind extract and grapefruit extract are provided as collagenase inhibitors. The collagenase inhibitor was obtained by mixing the red pomegranate rind extract and the grapefruit 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 inhibitors of examples 1 to 5 were dissolved in 5 sets of deionized water in volumes corresponding to those of comparative examples 1 to 5, respectively. Examples 1-5 test solutions of 5 sets of concentrations corresponding to comparative examples 1-5 were obtained. Wherein, the content (mass%) of the pomegranate rind extract and the grapefruit extract in the collagenase inhibitor is shown in table 1 below, and the logarithmic mass concentration of the collagenase inhibitor is shown in table 3 below.

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 and immediately mixing the materials, preserving the heat in a water bath at 37 ℃ for 10min, then adding 1mL (6.5%, w/v) of a trichloroacetic acid solution and mixing the materials uniformly, standing the mixture for 10min, centrifuging the mixture for 5min at 10000rpm, taking 0.5mL of a supernatant sample in the test tube into another test tube, adding 0.5mL (mass concentration: 10%) of a sodium bicarbonate test solution into the 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 inhibition rates of the red pomegranate rind extract (comparative examples 1 to 5) and the grapefruit extract (comparative examples 6 to 10) on the collagenase activity were calculated, respectively. 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 grapefruit 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 relationship diagram (figure 3), and calculating the mass concentration (IC) of the pomegranate bark extract when the red pomegranate bark extract and the grapefruit extract have the composite effect to generate the equivalent inhibition ratio (50 percent) by conversion_50a) Mass concentration of grapefruit extract ((IC) when the combined effect of pomegranate rind extract and grapefruit extract produces equivalent inhibition (50%))_50b) The results are shown in Table 3.

The effect of the combined action of the red pomegranate rind extract and the grapefruit 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_50Brepresents the mass concentration corresponding to the inhibition ratio of 50% of the grapefruit extract;

IC_50arepresents the mass concentration of the pomegranate bark extract when the compound action of the pomegranate bark extract and the grapefruit extract generates equivalent inhibition rate (50%);

IC_50brepresents the mass concentration of the grapefruit extract when the combined action of the pomegranate rind extract and the grapefruit extract produces an equivalent inhibition rate (50%).

Wherein γ ═ 1, indicates that the red pomegranate rind extract and grapefruit extract exhibit a simple additive effect; gamma is less than 1, the pomegranate rind extract and the grapefruit 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 grapefruit 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 2 are embodied in the present invention as reference examples 1 and 2, which can be compared with examples 3 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 the red pomegranate rind extract and the grapefruit extract may exhibit excellent synergistic effects.

Application examples 1 to 5

The firming cream is prepared according to the following production process steps and the contents (mass percent) of the components in the firming cream formulas of the application examples 1 to 5 in the following table 4. The production process comprises the following steps:

1. adding the phase A raw material into an oil phase pot, stirring and heating to 80-85 ℃ to fully dissolve the phase A raw material;

2. adding phase B into an emulsifying pot, stirring and heating to 80-85 ℃ to fully dissolve;

3. slowly pumping the oil phase substances in the oil phase pot into an emulsifying pot, stirring, homogenizing, vacuum emulsifying, and keeping the temperature of the emulsifying pot at 80-85 deg.C;

4. cooling to 42 deg.C, adding phase C and phase D, and stirring;

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

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

Phase A: isopropyl myristate, cyclomethicone, caprylic/capric triglyceride, oleyl erucate, cyclohexasiloxane, polyglyceryl-3-methylglucose distearate, shea butter, dimethicone, a complex of polyacrylamide and laureth-7 and C13-14 isoparaffins, C12-15 alcohol benzoate, a complex of PEG-100 stearate and glyceryl stearate, ceramide 2, tocopheryl acetate, olive oil unsaponifiable, glyceryl polyacrylate, ammonium acryloyldimethyl taurate/VP copolymer, stearyl glycyrrhetinate, butylated hydroxytoluene, methyl hydroxybenzoate, propyl hydroxybenzoate, perfume;

phase B: water, glycerin, propylene glycol, panthenol, betaine, aminomethyl propanol, colloidal oat flour, allantoin, carbomer, manganese chloride;

and C phase: polyethylene glycol-90M, butanediol;

phase D: pomegranate rind extract, grapefruit extract, phenoxyethanol, lactobacillus/soybean fermentation product extract, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate.

Wherein, glycerin, panthenol, betaine, glycerin polyacrylate and butanediol are used as moisturizers;

isopropyl myristate, cyclomethicone, caprylic/capric triglyceride, oleyl erucate, cyclohexasiloxane, shea butter, dimethicone, C12-15 alcohol benzoate, and olive oil unsaponifiable matter are oils and fats;

polyethylene glycol-90M, acryloyl dimethyl ammonium taurate/VP copolymer and carbomer are thickening agents;

pomegranate rind extract, grapefruit extract and collagenase inhibitor;

polyglyceryl-3-methylglucose distearate, a complex of polyacrylamide and laureth-7 and C13-14 isoparaffins, a complex of PEG-100 stearate and glyceryl stearate are emulsifiers;

ceramide 2, lactobacillus/soybean fermentation product extract, colloidal oat flour, allantoin, and manganese chloride are skin conditioners;

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

stearyl glycyrrhetinate is a soothing agent; butylated hydroxytoluene and tocopherol acetate are antioxidants;

phenoxyethanol, methyl hydroxybenzoate, propyl hydroxybenzoate as antiseptic;

aminomethyl propanol is a pH adjusting agent; the essence is an aromatic.

PEG-100 stearate and glyceryl stearate, manufacturer, procto, trade mark: ARLACEL 165;

the manufacturer of the polyacrylamide and laureth-7 and C13-14 isoparaffin complex is Sebik under the name Sepigel 305.

Application of comparative examples 1 to 8

The firming cream was prepared according to the contents (mass percentages) of the components in the firming cream formulations of the comparative application examples 1 to 8 in the following table 5 in the same manner as in the application examples 1 to 5.

TABLE 4

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 is 33, the test period is 8 weeks, the test selects the firming cream of the application examples 1 to 5 and the firming cream of the application comparative examples 1 to 8 as test samples, 13 different areas are divided on the forearm of the subject, the firming cream of the application examples 1 to 5 and the firming cream of the application comparative examples 1 to 8 are smeared on different areas on the inner side of the forearm every morning and evening, and the smearing amount is 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 rind extract and the grapefruit 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. The firming cream is characterized by comprising a collagenase inhibitor and a penetration enhancer, wherein the collagenase inhibitor is added in an amount of 0.01-10% of the total mass of the firming cream; the addition amount of the penetration enhancer is 0.01-10%;

2. The tightening cream according to claim 1, characterized in that the collagenase is a interstitial collagenase.

3. The tightening cream according to claim 1 or 2, wherein the mass ratio of the red pomegranate rind extract to the grapefruit extract is 1: 0.6-5.5, preferably 1: 0.8-5, more preferably 1: 1-4.5, further preferably 1: 1.2-4, further preferably 1: 1.5-3.5, and still further preferably 1: 1.8-3.

4. The tightening cream of any one of claims 1-3, 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.

5. The tightening cream according to any one of claims 1-4, wherein the tightening cream further comprises one or a combination of two or more of a moisturizer, a thickener, a pH adjuster, a grease, a preservative, a skin conditioner, an emulsifier, a soothing agent, a chelating agent, a fragrance, and an antioxidant; wherein the content of the first and second substances,

6. The tightening cream of claim 5, wherein the skin conditioning agent comprises one or a combination of two or more of manganese chloride, tapioca starch, ceramide 2, lactobacillus/soy fermentation product extract, palmitoyl tripeptide-5, acetyl hexapeptide-8, palmitoyl oligopeptide, colloidal oat flour, hydrolyzed collagen, hydrolyzed wheat protein, kelp extract, allantoin, pansy extract, chlorella fermentation product.

7. The tightening cream according to claim 5 or 6, wherein the soothing agent comprises one or a combination of two or more of Hamamelis virginiana water, stearyl glycyrrhetinate, bisabolol, ginger root extract, and aloe extract.

8. A method of preparing the firming cream as claimed in any one of claims 1 to 7, comprising the step of mixing the components of the firming cream.

9. A collagenase inhibitor comprising: the composition comprises a red pomegranate rind extract and a grapefruit extract, wherein the addition amount of the red pomegranate rind extract is 15-65% and the addition amount of the grapefruit extract is 35-85% based on the total mass of the collagenase inhibitor.

10. The collagenase inhibitor according to claim 9, wherein the mass ratio of the red pomegranate rind extract to the grapefruit extract is 1:0.6 to 5.5, preferably 1:0.8 to 5, more preferably 1:1 to 4.5, further preferably 1:1.2 to 4, further preferably 1:1.5 to 3.5, and further preferably 1:1.8 to 3.