CN113101252A

CN113101252A - Collagenase inhibitor, moisturizing lotion containing collagenase inhibitor and preparation method of moisturizing lotion

Info

Publication number: CN113101252A
Application number: CN202010031545.4A
Authority: CN
Inventors: 谢佩佩; 杨登亮; 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: 2020-01-13
Filing date: 2020-01-13
Publication date: 2021-07-13

Abstract

The invention provides a collagenase inhibitor, a moisturizing lotion containing the collagenase inhibitor and a preparation method of the moisturizing lotion. The collagenase inhibitor comprises: the collagenase inhibitor comprises a dendrobium officinale extract and a chamomile extract, wherein the adding amount of the dendrobium officinale extract is 17-65% and the adding amount of the chamomile extract is 35-83% 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, moisturizing lotion containing collagenase inhibitor and preparation method of moisturizing lotion

Technical Field

The invention relates to a collagenase inhibitor, a moisturizing crystal lotion containing the collagenase inhibitor and a preparation method of the moisturizing crystal lotion, 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 invention also provides a moisturizing lotion which contains collagenase inhibitor and has excellent anti-aging effect.

Furthermore, the invention also provides a preparation method of the moisturizing crystal lotion, 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 dendrobium officinale extract and a chamomile extract, wherein the adding amount of the dendrobium officinale extract is 17-65% and the adding amount of the chamomile extract is 35-83% based on the total mass of the collagenase inhibitor.

The collagenase inhibitor comprises the dendrobium officinale extract and chamomile extract, wherein the mass ratio of the dendrobium officinale extract to the chamomile extract is 1: 0.55-4.8, preferably 1: 0.58-4.5, more preferably 1: 0.6-4.2, further preferably 1: 0.62-4, further preferably 1: 0.65-3.8, and further preferably 1: 0.7-3.5.

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

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

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

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

The moisturizing crystal lotion further comprises one or a combination of more than two of a humectant, a thickening agent, a pH regulator, a preservative, a skin conditioner, a solubilizer, an aromatic and a soothing agent;

preferably, the addition amount of the humectant is 0.01-20%; 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 skin conditioner is 0.01-5%; the addition amount of the solubilizer is 0.01-0.5%; the adding amount of the aromatic is 0.005-0.5%; the addition amount of the allergy relieving agent is 0.01-5%.

The moisturizing lotion comprises one or more of hydrolyzed collagen, oat peptide, ceramide 3, Phaeodactylum tricornutum extract, Macrocystis japonica extract, Fucus vesiculosus extract, chlorella vulgaris fermentation product, Chlorella extract, Rhodophyta extract, brown algae extract, allantoin, Viscum album Biloba leaf extract, lalang grass rhizome extract, serine, cactus extract, Chondrus crispus extract and Pleurotus antarctica extract.

The moisturizing crystal comprises hamamelis water, centella asiatica extract, ginger root extract, bisabolol, dipotassium glycyrrhizinate and stearyl glycyrrhetinate, wherein the soothing agent is one or the combination of more than two of the hamamelis water, the centella asiatica extract and the ginger root extract.

The invention further provides a preparation method of the moisturizing lotion, which comprises the step of mixing the components of the moisturizing lotion.

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

The preparation method of the moisturizing crystal dew 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 ratio of the Dendrobium officinale extract of comparative examples 1-5 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 Dendrobium officinale extract in collagenase inhibitors according to examples 1-5 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 dendrobium officinale extract and a chamomile extract, wherein the adding amount of the dendrobium officinale extract is 17-65% and the adding amount of the chamomile extract is 35-83% based on the total mass of the collagenase inhibitor.

The inventor of the present invention found that an excellent synergistic effect can be produced using a combination of chamomile extract and dendrobium officinale extract, so that collagenase activity can be inhibited to achieve an anti-aging effect.

Specifically, in the invention, the mass ratio of the dendrobium officinale extract to the chamomile extract is 1: 0.55-4.8, preferably 1: 0.58-4.5, more preferably 1: 0.6-4.2, further preferably 1: 0.62-4, further preferably 1: 0.65-3.8, and further preferably 1: 0.7-3.5. When the mass ratio of the dendrobium officinale extract to the chamomile extract is within the above range, a synergistic effect can be further achieved, 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 dendrobium officinale extract and the chamomile extract are uniformly mixed by adopting a conventional mixing mode.

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.

Dendrobium officinale extract

Dendrobium officinale (Dendrobium officinale) is a perennial epiphytic herb of the genus Dendrobium of the family Orchidaceae. The stem of Dendrobium officinale mainly contains water-soluble polysaccharide, which is a kind of immunopotentiator.

The dendrobium officinale extract is originally prepared from the stem of dendrobium officinale. Generally, dendrobium officinale can not only moisten skin, enhance skin elasticity, reduce skin wrinkles and prevent skin aging. The herba Dendrobii extract also has effects of relieving fatigue, relieving mental stress, supplementing physical strength, nourishing brain and strengthening body constitution. In addition, the dendrobium officinale extract can also promote metabolism and enhance function, and has the effects of moistening lung, clearing intestinal heat and tonifying lung deficiency.

In the invention, the adding amount of the dendrobium officinale extract is 17-65% of the total mass of the collagenase inhibitor. When the addition amount of the dendrobium officinale extract is 17-65%, the activity of collagenase can be effectively inhibited. Specifically, the addition amount of the dendrobium officinale extract can be 20%, 25%, 30%, 32%, 35%, 40%, 45%, 50%, 55%, 60% and the like.

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 35-83% by mass based on the total mass of the collagenase inhibitor. When the addition amount of the chamomile extract is 35-83%, collagenase activity can be effectively inhibited. Specifically, the added amount of the chamomile extract may be 40%, 45%, 50%, 55%, 60%, 65%, 70%, 72%, 75%, 80%, or the like.

A second aspect of the invention provides a moisturizing lotion comprising a collagenase inhibitor according to the first aspect of the invention and a penetration enhancer; the appropriate content of collagenase inhibitor is added, so that the activity of collagenase, particularly the activity of interstitial collagenase can be inhibited, the moisturizing lotion 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 into the skin, the moisturizing lotion of the invention also uses penetration enhancers. By using a penetration enhancer, the collagenase inhibitor of the present invention can be exerted to a greater extent.

Wherein, the adding amount of the collagenase inhibitor is 0.01-10% of the total mass of the moisturizing lotion, 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 moisturizing crystal lotion, such as: 0.5%, 1%, 2%, 3%, 5%, 6%, 7%, 8%, etc. 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 to 5% and the amount of bis-diethoxydiol cyclohexane 1, 4-dicarboxylate added is 0.01 to 5% based on the total mass of the moisturizing gel. 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%, etc.

In the invention, the moisturizing crystal further comprises one or a combination of more than two of a humectant, a thickening agent, a pH regulator, a preservative, a skin conditioner, a solubilizer, a fragrance and a soothing agent; the formula of the moisturizing crystal lotion is mild, so that the efficacy of the collagenase inhibitor can be fully exerted. Specifically, the method comprises the following steps:

the adding amount of the humectant is 0.01-20% of the total mass of the moisturizing crystal lotion. When the addition amount of the humectant is 0.01-20%, the humectant can play a role in keeping moisture. In order to further exert the efficacy of the humectant, the humectant can be added in an amount of 1-20%, 3-20%, 5-19.5%, 6-19.5%, 8-19%, etc.

In the invention, the humectant comprises one or a combination of more than two of glycerin, dipropylene glycol, glyceryl polyether-26, medium molecular weight sodium hyaluronate, small molecular weight sodium hyaluronate, panthenol, PEG/PPG-17/6 copolymer, butanediol, PEG-32, PEG-8, PCA sodium, xylitol, erythritol, glycerin polyacrylate, mannose and trehalose. Wherein the molecular weight range of the medium molecular weight sodium hyaluronate is 500 KDa-less than 2000 KDa; the molecular weight range of the small molecular weight sodium hyaluronate is 10KDa to less than 500 KDa.

Further, in the moisturizer, one or a combination of two or more of glycerin, dipropylene glycol, glyceryl polyether-26, PEG/PPG-17/6 copolymer, mannose, hyaluronic acid, medium molecular weight sodium hyaluronate, small molecular weight sodium hyaluronate, panthenol, xylitol, erythritol, PEG-8, sodium PCA, glycerin polyacrylate, PEG-32, butylene glycol, and the like are preferably added, and the moisturizing effect can be further improved.

The addition amount of the thickening agent is 0.02-0.8% of the total mass of the moisturizing crystal lotion. By adding the thickening agent, the moisturizing lotion can be more textured. Preferably, the thickener of the present invention may be added in an amount of 0.05 to 0.6%, 0.1 to 0.5%, or the like.

In the invention, the thickening agent comprises one or the combination of more than two of carbomer, xanthan gum, sclerotium rolfsii gum, polydimethylsiloxane/vinyl polydimethylsiloxane cross-linked polymer, behenyl alcohol, polyacrylamide, PEG-100 stearate, acryloyl dimethyl taurate/VP copolymer and hydroxyethyl acrylate/acryloyl dimethyl taurate copolymer.

In the invention, a proper amount of sclerotium rolfsii gum can be added, and the sclerotium rolfsii gum is a novel non-ionic salt-resistant and temperature-resistant biological polysaccharide polymer-scleroglucan produced by fungi sclerotium rolfsii. Scleroglucan (scleroglucan) is a natural, encapsulated glucan (Capsular polysarido). The scleroglucan solution also has excellent stability at high temperatures, good applicability over a wide pH range, and great tolerance to various electrolytes present in the solution. In addition, the sclerotium rolfsii gum can also be used for ensuring that the moisturizing crystal dew has excellent water replenishing effect.

The addition amount of the pH regulator is 0.01-1% of the total mass of the moisturizing crystal dew. The pH value of the moisturizing crystal lotion 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 addition amount of the pH adjuster is more than 1% or less than 0.01%, a moisturizing lotion with a proper pH value cannot be obtained.

In the present invention, the pH adjuster includes one or a combination of two or more of aminomethyl propanol, arginine, citric acid, sodium citrate, sodium hydroxide, and the like.

The addition amount of the solubilizer is 0.01-0.5% based on the total mass of the moisturizing crystal lotion. By adding the solubilizer, the raw materials of the moisturizing crystal lotion can be dissolved more easily. Preferably, the solubilizer is added in an amount of 0.05-0.3%, 0.1-0.2%, etc. In the present invention, the solubilizer includes one or more of polysorbate-20, PEG-40 hydrogenated castor oil, and glyceryl ether-25 PCA isostearate.

In order to further improve the efficacy of the moisturizing lotion, the moisturizing lotion further comprises the skin conditioner. The skin conditioner is added to calm the skin, so that the facial skin can be relieved from injury and redness and swelling, and wrinkles can be reduced. In addition, the moisturizing crystal lotion can improve the skin oil-yielding condition, is not greasy in texture, and can adjust the water-oil balance.

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

In the present invention, the skin conditioner includes one or a combination of two or more of hydrolyzed collagen, avenin, ceramide 3, phaeodactylum tricornutum extract, macroalgae extract, fucus extract, chlorella fermentation product, green algae extract, rhodophyta extract, brown algae extract, allantoin, mistletoe ginkgo leaf extract, cogongrass rhizome extract, serine, cactus extract, Chondrus crispus extract, Pleurotus antarctica extract.

The moisturizing crystal lotion 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 soothing agent is 0.01-5% of the total mass of the moisturizing crystal lotion; 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 soothing agent comprises Hamamelis virginiana flower water, herba Centellae extract, rhizoma Zingiberis recens extract, bisabolol, dipotassium glycyrrhizinate, and stearyl glycyrrhetinate.

In addition, the moisturizing crystal dew also comprises a preservative and an aromatic. The addition amount of the preservative is 0.01-1.5% based on the total mass of the moisturizing crystal lotion; the addition amount of the aromatic is 0.005-0.5%. 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.

The moisturizing crystal lotion disclosed by the invention not only can play a role in resisting aging, but also can supplement water and moisturize, and the water wettability is very good. Therefore, the moisturizing crystal lotion can greatly improve the skin state, delay skin aging and be moist and glossy.

A third aspect of the present invention provides a method for preparing the moisturizing lotion of the second aspect, comprising a step of mixing the components of the moisturizing lotion.

Specifically, the preparation method of the moisturizing crystal lotion can comprise the following steps:

1. adding water, humectant, thickener, partial penetration enhancer, and partial antiseptic into a stirring pot, stirring, and heating to 75-85 deg.C;

2. cooling to 40-45 deg.C, adding pH regulator, solubilizer, aromatic, collagenase inhibitor, residual penetration enhancer, skin conditioner, soothing agent and residual antiseptic, and stirring;

3. cooling to 35-40 deg.C, discharging after qualified inspection, and standing for 12-48 hr;

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

Examples

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

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

The dendrobium officinale extract is purchased from: quzhou City exhibition-macro Biotechnology Ltd.

Comparative examples 1 to 5

Herba Dendrobii extract is provided as collagenase inhibitor. Dissolving the dendrobium officinale 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 logarithmic mass concentration of the dendrobium officinale extract is shown in the following table 2 or figure 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 then as shown in table 2 or figure 2 below.

Examples 1 to 5

Herba Dendrobii extract and flos Matricariae Chamomillae extract are provided as collagenase inhibitor. The collagenase inhibitor is obtained by mixing the dendrobium officinale extract and the chamomile extract according to the mass ratio of 3:1 (example 1), 2:1 (example 2), 1:1 (example 3), 1:2 (example 4) and 1:3 (example 5).

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

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

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

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

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

Wherein, the contents (mass%) of the dendrobium officinale extract and the chamomile extract in the collagenase inhibitors are shown in the following table 1, and the logarithmic mass concentration of the collagenase inhibitors is shown in the following table 3.

TABLE 1

Collagenase activity inhibition assay

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

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

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

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 inhibition rates of the dendrobium officinale extract (comparative examples 1-5) and the chamomile extract (comparative examples 6-10) on the activity of the collagenase are respectively calculated. And calculating the corresponding mass concentration (IC) when the inhibition rate of herba Dendrobii extract is 50% by combining the logarithmic mass concentration-collagenase inhibition rate relationship diagram (figure 1 and figure 2)_50A) Mass concentration (IC) corresponding to 50% inhibition ratio of chamomile extract_50B) The results are shown in Table 2.

TABLE 2

The collagenase inhibitors of examples 1-5 were then tested for collagenase inhibition. And calculating the mass concentration (IC) of the Dendrobium officinale extract when the composite action of the Dendrobium officinale extract and the chamomile extract generates equivalent inhibition rate (50%) by combining with a relation graph (figure 3) of logarithmic mass concentration-collagenase inhibition rate_50a) When the dendrobium officinale extract and the chamomile extract have combined action to generate equivalent inhibition rate (50%), the mass concentration of the chamomile extract ((IC)_50b) The results are shown in Table 3.

The effect of the combined effect of the dendrobium officinale extract and the chamomile extract can be evaluated by using an interaction coefficient gamma, and the specific results are shown in table 3.

γ＝IC_50a/IC_50A+IC_50b/IC_50B

Wherein, IC_50ARepresenting the corresponding mass concentration when the inhibition rate of the dendrobium officinale 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 dendrobium officinale extract is shown when the dendrobium officinale extract and the chamomile extract have combined action to generate equivalent inhibition rate (50%);

IC_50bthe mass concentration of the chamomile extract is shown when the dendrobium officinale extract and the chamomile extract have combined action to generate equivalent inhibition rate (50%).

Wherein, gamma is 1, which indicates that the dendrobium officinale extract and the chamomile extract show simple addition effect; gamma is less than 1, the dendrobium officinale extract and the chamomile extract show a synergistic effect, and the smaller the gamma value is, the stronger the synergistic effect is; the gamma is more than 1, the dendrobium officinale extract and the chamomile extract show antagonistic effect, and the larger the gamma value is, the larger the antagonistic effect is.

TABLE 3

Note: in Table 3, examples 1-2 are embodied in the present invention as reference examples 1-2, which can be contrasted with examples 3-5.

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 below 0.9, and even below 0.8, so that the dendrobium officinale extract and the chamomile extract may exhibit excellent synergistic effects.

Application examples 1 to 5

The moisturizing essence lotion is prepared according to the content (mass percentage) of each component in the moisturizing essence lotion formula 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 A, B and C phase raw materials into a stirring pot, stirring and heating to 83 ℃;

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

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

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

Phase A: water;

phase B: butanediol, propylene glycol, glycerol, medium molecular weight sodium hyaluronate, panthenol, dipropylene glycol, erythritol, xylitol, carbomer, sclerotium rolfsii, small molecular weight sodium hyaluronate and glycerol polyacrylate;

and C phase: methylparaben, mannose, PEG-8, PEG-32, sodium PCA, glyceryl polyether-26, PEG/PPG-17/6 copolymer;

phase D: aminomethyl propanol;

phase E: PEG-40 hydrogenated castor oil, essence;

and (3) phase F: chamomile extract, dendrobium officinale extract, clitocybe antarctica extract, carrageen crispus extract, serine, dipotassium glycyrrhizinate, phenoxyethanol, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate.

Wherein butanediol, glycerol, medium molecular weight sodium hyaluronate, panthenol, dipropylene glycol, erythritol, xylitol, small molecular weight sodium hyaluronate, glycerol polyacrylate, mannose, PEG-8, PEG-32, sodium PCA, glyceryl polyether-26, and PEG/PPG-17/6 copolymer are moisturizers;

carbomer and sclerotium rolfsii gum are thickening agents; aminomethyl propanol is a pH adjusting agent;

the Antarctic Pleurotus citrinopileatus extract, Chondrus crispus extract, and serine are skin conditioner;

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

dipotassium glycyrrhizinate is a soothing agent; the essence is an aromatic;

PEG-40 hydrogenated castor oil is a solubilizer; methyl hydroxybenzoate and phenoxyethanol are antiseptic;

the flos Matricariae Chamomillae extract and herba Dendrobii extract are collagenase inhibitors.

Wherein the molecular weight range of the medium molecular weight sodium hyaluronate is 500 KDa-less than 2000 KDa; the molecular weight range of the small molecular weight sodium hyaluronate is 10KDa to less than 500 KDa.

Application of comparative examples 1 to 8

The moisturizing essence lotion was prepared according to the contents (mass percentages) of the components in the moisturizing essence lotion 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 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 subjects is 33The test period is 8 weeks, the moisturizing essence lotions of application examples 1 to 5 and the moisturizing essence lotions of application comparative examples 1 to 8 are selected as test samples in the test, 13 different areas are divided on the forearm of a subject, the moisturizing essence lotions of application examples 1 to 5 and the moisturizing essence lotions of application comparative examples 1 to 8 are applied to different areas on the inner side of the forearm every morning and evening, and the applying 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 skin elasticity, i.e., enhanced skin elasticity, and thus, the use of chamomile extract and dendrobium officinale 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 dendrobium officinale extract and a chamomile extract, wherein the adding amount of the dendrobium officinale extract is 17-65% and the adding amount of the chamomile extract is 35-83% based on the total mass of the collagenase inhibitor.

2. The collagenase inhibitor according to claim 1, wherein the mass ratio of the dendrobium officinale extract to the chamomile extract is 1: 0.55-4.8, preferably 1: 0.58-4.5, more preferably 1: 0.6-4.2, further preferably 1: 0.62-4, further preferably 1: 0.65-3.8, and still further preferably 1: 0.7-3.5.

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 the collagenase inhibitor.

5. A moisturizing lotion, characterized by comprising the collagenase inhibitor and the penetration promoter according to any one of claims 1 to 3, wherein the collagenase inhibitor is added in an amount of 0.01 to 10 percent based on the total mass of the moisturizing lotion; the addition amount of the penetration enhancer is 0.01-10%.

6. The moisturizing lotion of claim 5, 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.

7. The moisturizing lotion of claim 5 or 6, further comprising one or a combination of two or more of a humectant, a thickener, a pH adjuster, a preservative, a skin conditioner, a solubilizer, a fragrance, and a soothing agent;

8. The moisturizing lotion of claim 7, wherein the skin conditioner comprises one or a combination of two or more of hydrolyzed collagen, avenin, ceramide 3, Phaeodactylum tricornutum extract, Macrocystis extract, Fucus vesiculosus extract, Chlorella fermentation product, Chlorella extract, Rhodophyta extract, Brown algae extract, allantoin, Viscum album Biloba leaf extract, Imperata rhizome extract, serine, Opuntia ficus extract, Chondrus crispus extract, Pleurotus antarctica extract.

9. The moisturizing lotion of claim 7 or 8, wherein the soothing agent comprises one or a combination of two or more of hamamelis water, centella asiatica extract, ginger root extract, bisabolol, dipotassium glycyrrhizinate, and stearyl glycyrrhetinate.

10. A method of preparing a moisturizing lotion according to any of claims 5-9, comprising the step of mixing the components of the moisturizing lotion.