Propolis liposome gel as well as preparation method and application thereof

Abstract

The invention discloses propolis liposome gel as well as a preparation method and application thereof. The propolis gel comprises propolis liposome, a glycyrrhiza glabra root extract, an aloe barbadensis leaf extract, a centella asiatica extract, a snow lotus herb extract, a Chinese herbaceous peony root extract, a purslane herb extract and gel acceptable auxiliary materials. The preparation method of the propolis liposome comprises the following steps: taking propolis, crushing, extracting with a cosolvent, filtering to obtain a propolis liquid, adding a dispersion medium into the propolis liquid, and continuously reacting to obtain the propolis liposome. The dispersion medium is composed of phosphatidyl ethanolamine, a weakly acidic anionic surfactant disodium lauryl sulfosuccinate and water. The product disclosed by the invention is easily absorbed by an organism, contains rich bioactive substances, is combined with plant extracts with the effects of whitening, beautifying and the like, can remarkably improve the dark yellow condition of skin, improve the brightness of the skin and improve the color of the skin, and is a functional gel product suitable for various skin types.

Description

Technical Field

[0001] The invention belongs to the field of skin care, and in particular relates to a propolis liposome gel and a preparation method and application thereof. Background Art

[0002] Melanin is mainly produced by melanocytes in the basal layer of the human epidermis. Its content and distribution determine the depth of human skin color and are essential for protecting the skin from ultraviolet radiation and free radicals that damage DNA. However, its excessive accumulation not only causes aesthetic problems, but also induces pigmentation disorders, causing chloasma, freckles, age spots, etc., affecting people's quality of life. Among them, melasma is a common chronic, acquired, hyperpigmented skin disease with an incidence rate of up to 30% in Asian women of childbearing age. The main clinical manifestation is light brown to dark brown patches with clear boundaries on the face. Its pathogenesis is too complicated, and patients often show different clinical and histological characteristics. Therefore, the clinical treatment effect is poor and it is very easy to relapse. There is an urgent need for a portable, safe and effective treatment plan.

[0003] In recent years, foreign countries have attached great importance to the application of propolis in the cosmetics industry, given that propolis has multiple functions such as antiseptic, antibacterial, anti-inflammatory, antipruritic, deodorant, sunscreen, skin care and tissue regeneration. Studies have shown that propolis has whitening effects, anti-skin aging, anti-free radical oxidation and the function of inhibiting tyrosinase activity, can reduce the production of melanin in the skin, absorb ultraviolet rays, and has the effect of eliminating chloasma, with a definite therapeutic effect.

[0004] Chinese patent CN115300444B discloses a preparation method and application of propolis liposomes, wherein the method dissolves propolis in a butylene glycol organic medium, then adds a dispersion medium, and reacts for at least one hour to obtain propolis liposomes, wherein the dispersion medium is composed of phosphatidylcholine, glycerol and water, and the mass ratio of the phosphatidylcholine, glycerol and water is (2.8-3.2): (0.8-1.2): (0.8-1.2), and most preferably 3: 1: 1. In the method, a large amount of glycerol is used in the dispersion medium, and the mass ratio of glycerol and water in the optimal ratio reaches 1: 1, but glycerol has strong hygroscopicity, especially when other active ingredients are added, glycerol may adversely affect the effect of other active ingredients, because the glycerol molecule contains a large amount of hydroxyl groups, and is easy to produce strong hydrogen bonds with other active ingredients. Therefore, when ensuring that the prepared liposome meets the requirements, it is an important research direction to minimize the amount of glycerol. In addition, although butylene glycol can be used in moisturizing agents, it is relatively expensive, and it is best to use cheaper solvents, such as ethanol, etc., which can effectively reduce production costs. Summary of the invention

[0005] In view of the problems existing in the prior art, according to one aspect of the present invention, an object of the present invention is to provide a propolis liposome, wherein the propolis liposome is prepared according to the following method:

[0006] Weigh propolis, crush it, add 4-5 times the volume of a solvent, extract for 12-36 hours, filter to obtain propolis liquid, add a dispersion medium to the propolis liquid, continue to react for 0.5 to 2 hours at 60-85°C and 500-600r / min of stirring conditions to obtain propolis liposomes.

[0007] Preferably, the co-solvent is an ethanol solution with a volume percentage concentration of 70 to 95%.

[0008] Preferably, the dispersion medium consists of phosphatidylethanolamine (cephalin PE), a weakly acidic anionic surfactant disodium lauryl sulfosuccinate and water.

[0009] Preferably, the mass ratio of the propolis liquid to the dispersion medium is 0.5:1 to 1.5:1, more preferably 0.8:1 to 1.2:1, and most preferably 1:1.

[0010] Preferably, the mass ratio of phosphatidylethanolamine, disodium lauryl sulfosuccinate and water is (4-6):(0.01-0.8):1; preferably (2.4-3):(0.04-0.5):1, and most preferably 2.4:0.08:1. Phosphatidylethanolamine is a weakly alkaline phospholipid, so the use of a certain amount of weakly acidic anionic surfactant disodium lauryl sulfosuccinate can not only better maintain the pH of the liposome, but also make it easier to maintain the stability of the liposome membrane structure and improve the storage stability of the liposome.

[0011] Preferably, the reaction temperature is 65 to 80° C., most preferably 70-75° C. The increase in reaction temperature helps to accelerate the formation process of propolis liposomes, so increasing the temperature as much as possible helps to shorten the reaction time and improve efficiency, but too high a temperature can easily cause the quality of propolis to deteriorate. Since the weakly acidic anionic surfactant disodium lauryl sulfosuccinate is a white and delicate paste at room temperature and a transparent liquid above 70° C., it is most appropriate to keep the reaction temperature above 70° C. as much as possible.

[0012] Preferably, the reaction time is 0.5 to 1.5 hours, most preferably 1 hour.

[0013] Preferably, the propolis liposomes according to the present invention do not contain glycerol components.

[0014] According to another aspect of the present invention, another object of the present invention is to provide a propolis gel comprising the following active ingredients in parts by weight:

[0015] 0.1-10 parts of propolis liposomes, 2-25 parts of licorice root extract, 1-25 parts of aloe vera leaf extract, 0.5-20 parts of Centella asiatica extract, 0.5-20 parts of snow lotus extract, 0.2-20 parts of peony root extract and 0.1-20 parts of purslane extract, as well as gel-acceptable excipients.

[0016] The acceptable auxiliary materials of the gel include 0.1-10 parts of gel matrix carbomer, 0.1-10 parts of pH value regulator, 0.05-2 parts of surfactant, and 20-75 parts of cosolvent.

[0017] Preferably, the propolis gel comprises the following active ingredients in parts by weight: 0.2-5 parts of propolis liposomes, 5-20 parts of Glycyrrhiza glabra root extract, 2-20 parts of Aloe barbadensis leaf extract, 1-15 parts of Centella asiatica extract, 1-15 parts of Snow Lotus extract, 0.5-15 parts of Paeonia lactiflora root extract and 0.5-15 parts of Portulaca oleracea extract.

[0018] Preferably, the acceptable auxiliary materials of the gel include 0.2-5 parts of gel matrix, 0.2-5 parts of pH regulator, 0.05-1 parts of surfactant, and 40-60 parts of cosolvent.

[0019] Preferably, the gel matrix is ​​carbomer.

[0020] Preferably, the surfactant is Tween 80.

[0021] Preferably, the pH regulator is triethanolamine.

[0022] Preferably, the co-solvent is ethanol with a volume percentage concentration of 70 to 95%.

[0023] Preferably, the propolis gel comprises the following active ingredients in parts by weight: 0.3-2.5 parts of propolis liposomes, 6-15 parts of Glycyrrhiza glabra root extract, 5-11 parts of Aloe vera leaf extract, 5-8 parts of Centella asiatica extract, 4-9 parts of Snow Lotus extract, 2-9 parts of Paeonia lactiflora root extract and 3-7 parts of Portulaca oleracea extract.

[0024] Preferably, the acceptable excipients of the gel include 0.8-3 parts of carbomer, 0.8-2 parts of triethanolamine, 0.3-1.5 parts of Tween 80, and 7-13 parts of ethanol with a volume percentage concentration of 70-95%.

[0025] More preferably, the propolis gel comprises the following active ingredients in parts by weight: 2.2-2.7 g of propolis liposomes, 13-17 g of Glycyrrhiza glabra root extract, 8-10 g of Aloe barbadensis leaf extract, 6-8 g of Centella asiatica extract, 4-6 g of Snow Lotus extract, 4-6 g of Paeonia lactiflora root extract and 3-5 g of Portulaca oleracea extract.

[0026] More preferably, the acceptable excipients of the gel include 2.8-3.2 g of carbomer, 1.1-1.3 g of triethanolamine, 0.2-0.4 g of Tween 80, and 8-12 g of ethanol with a volume percentage concentration of 90%.

[0027] According to another aspect of the present invention, another object of the present invention is to provide a method for preparing the propolis gel, the preparation method comprising the following steps:

[0028] (1) Preparation of plant extracts: Glycyrrhiza glabra root, Aloe vera leaf, Centella asiatica, Saussurea involucrata, Paeonia lactiflora root and Portulaca oleracea are mixed or extracted independently by percolation method, and concentrated to a relative density of 0.8-1.5 mg / ml at 60° C. to obtain respective plant extracts;

[0029] (2) Preparation of gel forming agent: mixing the gel matrix and part of the co-solvent, and fully swelling the mixture to obtain the gel forming agent;

[0030] (3) Add propolis liposomes and plant extracts to the gel forming agent, mix well, add the remaining surfactant and pH regulator in sequence, and then add the remaining cosolvent and purified water to a sufficient amount to obtain propolis liposome gel.

[0031] Preferably, during the percolation extraction process in step (1), the percolation solvent is 70-95% ethanol by volume, the mass ratio of the percolation solvent to the plant raw material is 1-20, and the percolation solvent flow rate is 1-15 ml / min.

[0032] More preferably, the percolation solvent is 80-95% ethanol, the volume mass ratio of the percolation solvent to the plant raw material is 1-15, and the percolation solvent flow rate is 2-10 ml / min. This extraction method can form a good concentration difference, has a high extraction efficiency, and room temperature extraction does not destroy the effective components of the plant raw material.

[0033] According to another aspect of the present invention, another object of the present invention is to provide the use of the propolis gel in the preparation of skin care products, wherein the skin care products include but are not limited to lotions, emulsions, cosmetic creams, facial masks, paste gels, sprays, facial cleansers, foundations, powder creams, body lotions or massage creams, etc.

[0034] Beneficial Effects

[0035] 1) The present invention selects to develop propolis liposomes. Liposomes are an artificially prepared drug delivery system that can encapsulate active ingredients in lipid bilayers to form micro-spherical carriers. Gel is a transparent or translucent semisolid preparation with a three-dimensional network structure. Dispersing a liposome suspension in a polymer gel matrix can form a new type of gel. Liposomes have the advantages of penetrating the stratum corneum, improving the local efficacy of drugs, and having little irritation to the skin. When combined with gel, they not only retain their respective advantages, but also form a multi-reservoir system, which enhances the retention time of drugs in the skin through the adhesion of the gel, exerts a long-acting sustained-release effect, reduces adverse drug reactions, reduces the number of medications, and is beneficial to local treatment of the skin.

[0036] 2) The present invention chooses to make a gel, which has good compatibility with plant extracts, is easy to spread, has good affinity with the skin and is non-irritating, can prolong the action time of the present invention on the skin, and helps the active ingredients of the present invention to be fully absorbed by the skin.

[0037] 3) Propolis is rich in a variety of active ingredients such as flavonoids, terpenes, organic acids, etc., and has a significant skin-soothing and muscle-promoting effect when applied externally. In the formula of the present invention, after the propolis liposomes are mixed with other plant extracts, auxiliary ingredients such as gel matrix, moisturizer, pH value regulator, surfactant and cosolvent are added thereto, and a gel product with stable performance is prepared through the synergistic effect between different components. This gel has good moisturizing properties, and the effective ingredients therein are evenly distributed. It can play a good skin-soothing and muscle-promoting effect when applied to the skin surface.

[0038] 4) The pH value of the human skin surface is weakly acidic, about 5.0 to 7.0. Experiments have shown that the present invention is uniform and fine, free of impurities, no odor, moderate pH value, non-toxic and non-irritating, and has no side effects.

[0039] 5) The product of the present invention is relatively easy to be absorbed by the body, and contains rich biologically active substances. Combined with plant extracts with whitening and beauty-enhancing effects, it can significantly improve the dullness of the skin, increase the brightness of the skin, and improve the color of the skin. It is a functional gel product suitable for all skin types. DETAILED DESCRIPTION

[0040] Hereinafter, the present invention will be described in detail. Before describing, it should be understood that the terms used in this specification and the appended claims should not be interpreted as being limited to the general meaning and dictionary meaning, but should be interpreted according to the meaning and concept corresponding to the technical aspects of the present invention on the basis of the principle that the inventor is allowed to appropriately define the terms for the best interpretation. Therefore, the descriptions presented here are only preferred examples for illustrative purposes and are not intended to limit the scope of the present invention, so that it should be understood that other equivalents or improvements can be obtained therefrom without departing from the spirit and scope of the present invention.

[0041] In this document, the terms "include", "including", "have", "contain" or any other similar terms are open conjunctions, which are intended to cover non-exclusive inclusions. For example, a composition or product containing multiple elements is not limited to the elements listed herein, but may also include other elements that are not explicitly listed but are generally inherent to the composition or product. In addition, unless otherwise explicitly stated, the term "or" refers to an inclusive "or" rather than an exclusive "or". For example, any of the following situations satisfies the condition "A or B": A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), and A and B are both true (or exist). In addition, in this document, the interpretation of the terms "include", "including", "have", and "contain" should be considered to have been specifically disclosed and simultaneously cover closed or semi-closed conjunctions such as "consisting of" and "consisting essentially of".

[0042] In this article, all features or conditions defined in the form of numerical ranges or percentage ranges are only for brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be deemed to have covered and specifically disclosed all possible secondary ranges and individual values ​​within the range, especially integer values. For example, the range description of "1 to 8" should be deemed to have specifically disclosed all secondary ranges such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc., especially secondary ranges defined by all integer values, and should be deemed to have specifically disclosed individual values ​​such as 1, 2, 3, 4, 5, 6, 7, 8, etc. within the range. Unless otherwise specified, the above interpretation method applies to all contents of the entire present invention, regardless of whether the range is broad or not.

[0043] If the quantity or other numerical value or parameter is expressed as a range, a preferred range or a series of upper and lower limits, it should be understood that all ranges consisting of any upper limit or preferred value of the range and the lower limit or preferred value of the range have been specifically disclosed herein, regardless of whether these ranges are disclosed separately. In addition, if a numerical range is mentioned herein, unless otherwise specified, the range should include its endpoints and all integers and fractions within the range.

[0044] In this document, numerical values ​​should be understood to have the accuracy of the number of significant digits of the numerical value, provided that the purpose of the invention can be achieved. For example, the number 40.0 should be understood to cover the range from 39.50 to 40.49. "Parts" involved in this application refer to parts by weight, and the unit is standard weight unit such as g, kg, etc.

[0045] The gel matrix carbomer is a copolymer of polyalkyl sucrose or polyalkyl pentaerythritol and acrylic acid cross-linked polymer, soluble in water, ethanol and glycerin, has strong hygroscopicity, is non-irritating to the skin, forms a transparent film after coating, and has good heat resistance and cold resistance. Carbomer is used as the gel matrix, the active ingredients are evenly dispersed, there is no stratification or hydration phenomenon, and the formed gel is fine and uniform, moderate in consistency, and has strong adhesion, which can prolong the action time of the active ingredients on the skin.

[0046] Preferably, in the gel of the present invention, the surfactant is Tween 80. Tween 80, also known as polysorbate 80, is widely used. In addition to being an emulsifier, it is also used as a solubilizer for certain poorly soluble drugs. In the present invention, Tween 80 can promote the dissolution of various plant extracts, reduce the precipitation of active ingredients, and enhance the efficacy of the present invention.

[0047] One of the features of the present invention is that glycerol is not used in the preparation of propolis liposomes. Glycerol (also known as glycerol) is a common cosmetic ingredient and is widely used in moisturizing products because of its hygroscopicity (ability to attract and retain moisture). It helps the skin stay soft and moisturized and reduces dryness. However, although glycerol is safe and effective in most cases, it may also have some negative effects when used under certain conditions, such as excessive hygroscopicity: the strong hygroscopicity of glycerol means that it can absorb moisture. Since the propolis molecules need to be "wrapped" by water molecules in order to maintain the activity of propolis in the propolis liposomes of the present invention, if glycerol is present, long-term storage may cause the inactivation of the propolis molecules. And if the environment is very dry (such as winter or low humidity areas), glycerol may absorb moisture from the inside of the skin, causing the skin to become drier. In addition, high concentrations of glycerol solutions can cause stinging or burning sensations when applied directly to the skin, especially for open wounds or extremely dry skin. The hygroscopic properties of glycerol may affect the stability or effects of other active ingredients, especially when the formula according to the present invention contains propolis active ingredients.

[0048] The following examples are only listed as examples of embodiments of the present invention and do not constitute any limitation to the present invention. It can be understood by those skilled in the art that modifications within the scope of the essence and concept of the present invention fall within the scope of protection of the present invention. Unless otherwise specified, the reagents and instruments used in the following examples are commercially available products.

[0049] Example 1

[0050] The propolis liposome gel of this embodiment is prepared from the following raw materials in parts by weight:

[0051] Propolis liposomes 2g, Glycyrrhiza glabra root extract 10g, Aloe barbadensis leaf extract 11g, Centella asiatica extract 5g, Snow lotus extract 9g, Paeonia lactiflora root extract 9g and Portulaca oleracea extract 5g;

[0052] Acceptable excipients for the gel include: 1 g of carbomer, 0.8 g of triethanolamine, 1.5 g of Tween 80, 9 g of ethanol with a volume percentage concentration of 90%, and purified water to make up the total amount to 100 g.

[0053] The method for preparing the product of this embodiment comprises the following steps:

[0054] (1) Preparation of propolis liposomes: weigh propolis, crush it, add 4-5 times the amount of co-solvent to extract for 24 hours, filter and obtain propolis liquid; add dispersion medium to the propolis liquid at 72° C., and continue the reaction for at least 1 hour to obtain the propolis liposomes; the mass ratio of the propolis liquid to the dispersion medium is 1:1; the dispersion medium is composed of phosphatidylethanolamine (cephalin PE), weakly acidic anionic surfactant disodium lauryl sulfosuccinate and water, and the mass ratio is 2.4:0.08:1;

[0055] (2) Preparation of plant extracts: Five plant raw materials, namely, root of Glycyrrhiza glabra, leaf of Aloe vera, Centella asiatica, Saussurea involucrata, root of Paeonia lactiflora and Portulaca oleracea, were mixed or extracted separately by percolation method, the percolation solvent was 95% alcohol by volume, the mass ratio of percolation solvent to plant raw material was 10:1, the percolation solvent flow rate was 4 ml / min, and the mixture was concentrated to a relative density of 1.20 mg / ml at 60° C. to obtain a plant extract;

[0056] (3) Preparation of a gel-forming agent: Carbomer, 0.8 g of part of triethanolamine and Tween were mixed, and part of ethanol with a volume percentage concentration of 90% was added, and after sufficient swelling, a gel-forming agent was obtained;

[0057] (4) Add propolis liposomes and plant extracts to the gel forming agent, mix well, add the remaining Tween 80 and triethanolamine in sequence, and then add the remaining ethanol with a volume percentage concentration of 90% and purified water to a sufficient amount to obtain propolis liposome gel.

[0058] Example 2

[0059] The propolis liposome gel of this embodiment is prepared from the following raw materials in parts by weight:

[0060] Propolis liposomes 2.5g, Glycyrrhiza glabra root extract 15g, Aloe barbadensis leaf extract 9g, Centella asiatica 7g, Snow lotus extract 5g, Paeonia lactiflora root extract 5g and Portulaca oleracea extract 4g;

[0061] Acceptable excipients for the gel include: 3 g of carbomer, 1.2 g of triethanolamine, 0.3 g of Tween 80, 10 g of 90% ethanol, and purified water to make up the total amount to 100 g.

[0062] The preparation method is the same as that of Example 1.

[0063] Example 3

[0064] The propolis liposome gel of this embodiment is prepared from the following raw materials in parts by weight:

[0065] Propolis liposome 1g, Glycyrrhiza glabra root extract 11g, Aloe barbadensis leaf extract 10g, Centella asiatica extract 8g, Snow lotus extract 8g, Paeonia lactiflora root extract 3g and Portulaca oleracea extract 6g;

[0066] Acceptable excipients for the gel include: carbomer 0.8 g, triethanolamine 1.5 g, Tween 80 1 g, 90% ethanol 13 g, and purified water to make up the total amount to 100 g.

[0067] The preparation method is the same as that of Example 1.

[0068] Example 4

[0069] The propolis liposome gel of this embodiment is prepared from the following raw materials in parts by weight:

[0070] Propolis liposome extract 1.9g, Licorice root extract 6g, Aloe barbadensis leaf extract 9g, Centella asiatica extract 7g, Snow lotus extract 4g, Peony root extract 9g and Portulaca oleracea extract 7g:

[0071] Acceptable excipients for the gel include: carbomer 2g, triethanolamine 2g, Tween 80 0.5g, 90% ethanol 7g, and purified water to make up the total amount to 100g.

[0072] The preparation method is the same as that of Example 1.

[0073] Comparative Example 1 (Compared with Example 2, without adding propolis)

[0074] The gel of this comparative example is prepared from the following raw materials:

[0075] 15g of Licorice root extract, 9g of Aloe barbadensis leaf extract, 7g of Centella asiatica extract, 5g of Snow Lotus extract, 5g of Paeonia lactiflora root extract, and 4g of Portulaca oleracea extract;

[0076] Acceptable excipients for the gel include: 3 g of carbomer, 1.2 g of triethanolamine, 0.3 g of Tween 80, 10 g of 90% ethanol, and purified water to make up the total amount to 100 g.

[0077] Prepared by the following method:

[0078] (1) Preparation of plant extracts: Five plant raw materials, namely, root of Glycyrrhiza glabra, leaf of Aloe vera, Centella asiatica, Saussurea involucrata, root of Paeonia lactiflora and Portulaca oleracea, are mixed or extracted separately by percolation method, the percolation solvent is 95% alcohol by volume, the mass ratio of percolation solvent to plant raw material is 10:1, the percolation solvent flow rate is 4 ml / min, and the mixture is concentrated to a relative density of 1.20 mg / ml at 60° C. to obtain a plant extract;

[0079] (2) Preparation of gel forming agent: Carbomer, 0.8 g of part of triethanolamine and Tween were mixed, and part of ethanol with a volume percentage concentration of 90% was added, and after sufficient swelling, a gel forming agent was obtained;

[0080] (3) Add the plant extract to the gel forming agent, mix well, add the remaining Tween 80 and triethanolamine in sequence, and then add the remaining ethanol with a volume percentage concentration of 90% and purified water to a sufficient amount to obtain a gel without propolis.

[0081] Comparative Example 2 (Compared with Example 2, glycerol was added)

[0082] The gel of this comparative example is prepared from the following raw materials:

[0083] Propolis liposomes 2.5g, Glycyrrhiza glabra root extract 15g, Aloe barbadensis leaf extract 9g, Centella asiatica 7g, Snow lotus extract 5g, Paeonia lactiflora root extract 5g and Portulaca oleracea extract 4g;

[0084] Acceptable excipients for the gel include: 3 g of carbomer, 1.2 g of triethanolamine, 0.3 g of Tween 80, 10 g of 90% ethanol, and purified water to make up the total amount to 100 g.

[0085] Prepared by the following method:

[0086] (1) Preparation of propolis liposomes: weigh propolis, crush it, add 4-5 times the amount of co-solvent to extract for 24 hours, filter to obtain propolis liquid; add dispersion medium to the propolis liquid at 72° C., and continue the reaction for at least 1 hour to obtain the propolis liposomes; the mass ratio of the propolis liquid to the dispersion medium is 1:1; the dispersion medium is composed of phosphatidylethanolamine (cephalin PE), glycerol, weakly acidic anionic surfactant disodium lauryl sulfosuccinate and water, and the mass ratio is 2.4:1.2:0.08:1;

[0087] (2) Preparation of plant extracts: Five plant raw materials, namely, root of Glycyrrhiza glabra, leaf of Aloe vera, Centella asiatica, Saussurea involucrata, root of Paeonia lactiflora and Portulaca oleracea, were mixed or extracted separately by percolation method, the percolation solvent was 95% alcohol by volume, the mass ratio of percolation solvent to plant raw material was 10:1, the percolation solvent flow rate was 4 ml / min, and the mixture was concentrated to a relative density of 1.20 mg / ml at 60° C. to obtain a plant extract;

[0088] (3) Preparation of a gel-forming agent: Carbomer, 0.8 g of part of triethanolamine and Tween were mixed, and part of ethanol with a volume percentage concentration of 90% was added, and after sufficient swelling, a gel-forming agent was obtained;

[0089] (4) Add propolis liposomes and plant extracts to the gel forming agent, mix well, add the remaining Tween 80 and triethanolamine in sequence, and then add the remaining ethanol with a volume percentage concentration of 90% and purified water to a sufficient amount to obtain propolis liposome gel.

[0090] Experimental Example 1

[0091] 1. Characterization of propolis liposomes

[0092] Take a small amount of the propolis liposomes prepared in Examples 1 to 4 and Comparative Example 2, dilute with water, drip 1 drop of sample onto the surface of a copper mesh, vacuum dry, negatively stain with a 1% phosphotungstic acid solution for 30 seconds, and observe its microscopic morphology with a transmission electron microscope. The results show that the propolis liposomes are spherical, there is no aggregation between particles, and the distribution is relatively uniform. Take an appropriate amount of propolis liposomes, dilute with deionized water, and measure the particle size and polydispersity index (PDI) with a laser particle size analyzer. The results show that the particle size of the propolis liposomes is (143.7±1.74) nm, and the PDI is 0.092±0.06.

[0093] 2. Preparation and characterization of propolis liposome gel

[0094] The propolis liposome gel prepared in Examples 1 to 4 and Comparative Example 2 was diluted and observed under a transmission electron microscope. It was found that there was still gel residue, but it was also found that the liposomes were spherical and the particle size remained basically unchanged compared with that before being added to the gel. It is believed that the liposomes were not destroyed after being added to the gel. 1g of propolis liposome gel was dissolved in 10mL of water and the mixture was stirred at 3000r·min. -1Centrifuge for 5 minutes, draw the supernatant, and use a laser particle size analyzer to measure its particle size and PDI. The results show that the particle size of the propolis liposome gel is (148.27±2.90) nm, and the PDI is 0.132±0.01. After the propolis liposome gel prepared in Examples 1 to 4 and Comparative Example 2 was stored for 2 weeks at room temperature and pressure, the liposome morphology in the propolis liposome gel prepared in Comparative Example 2 gradually became damaged, and about 5%-8% (average about 6.8%) of the liposome morphology was destroyed, and as time went on, after 4 weeks, about 7%-12% (average about 8.9%) of the liposome morphology was destroyed, while the morphology of the propolis liposome gel prepared in Examples 1 to 4 remained intact.

[0095] 3. Human trial evaluation

[0096] (1) Subjects

[0097] A total of 120 subjects aged 20 to 45 years old, all of whom were indoor workers, were selected and divided into 6 groups, with 20 subjects in each group, of which 5 groups were experimental groups and 1 group was a control group. The exclusion criteria of the subjects met the requirements of GB 171492 "Diagnostic criteria and treatment principles for cosmetic contact dermatitis"; no serious systemic diseases, no immunodeficiency or autoimmune diseases; no active allergic diseases; no history of allergy to skin care cosmetics; no systemic use of hormone drugs and immunosuppressants within 1 month; not pregnant or lactating; no ethical contraindications; according to the Chardon grouping method, the skin color was between Ⅱ and Ⅳ.

[0098] (2) Test indicators

[0099] Skin melanin content (MI value): MI value represents the melanin index, and the value range is 0 to 999. The higher the measured value, the higher the melanin content in the skin. The measurement principle of Mexameter MX18 skin melanin and hemoglobin test probe is based on the principle of spectral absorption (RGB), and the melanin content in the skin is determined by measuring the reflection amount of light of a specific wavelength on human skin.

[0100] Skin whiteness (brightness) (L* value): L* value is white balance. The larger the value, the more the color tends to be white, and vice versa.

[0101] Skin brightness (dullness) (ITA° value): The ITA° value is the skin individual type angle, which is a numerical value related to the L* value and b* value that characterizes the brightness of the skin. The larger the ITA° value, the brighter the skin, and vice versa, the darker the skin.

[0102] (3) Test method

[0103] ①Control the temperature at 20℃±2℃ and the relative humidity at 55%±3%. After the tester cleans his face slightly, sit in this environment for 30 minutes before the test. Use the skin melanin and hemoglobin test probes to test the MI value of the zygomatic area of ​​the face of the test subject before using the sample; use the skin color difference test probe to test the L* value and ITA° value of the face of the test subject before using the sample; use the VISIA facial image analysis system to take a photo of the whole face of the test subject before using the sample, and try to avoid smile lines, reflective areas, moles and hair in the analysis area.

[0104] ② According to the MI value, L* value and ITA° value tested before the subjects used the samples, all subjects were divided into a control group and an experimental group to ensure that there was no significant difference in the initial values ​​of the subjects in the control group and the experimental group.

[0105] ③ After cleansing their faces every day, the subjects in the five experimental groups were treated with the propolis liposome gels prepared in Examples 1 to 4 and the gel prepared in Comparative Example 1 at 0.5 μL / cm 2 The test area was evenly coated with the product at a uniform density, once in the morning and once in the evening, while a control group did not use any product after cleansing.

[0106] ④ 1, 2 and 4 weeks after using the sample, the MI value, L* value and ITA° value of the subjects in the control group and the experimental group were respectively measured under the conditions described in step ①, and the measurement was performed 3 times each time, and the average value was taken. The full-face photos of the subjects in the experimental group were taken using the VISIA facial image analysis system.

[0107] (4) Data analysis

[0108] SPSS 17.0 software was used to perform statistical analysis on the data, and paired t-test was used to analyze the significance of the data. When p < 0.05, the difference was considered significant; when p < 0.01, the difference was considered extremely significant.

[0109] (5) Analysis of test results

[0110] Skin MI value

[0111] The test results of the skin MI value are shown in Table 2 below. As can be seen from Table 2, within the 4 weeks of the test, after the subjects used the propolis liposome gel prepared according to Examples 1 to 4 of the present application, the MI value showed a significant downward trend, and at the 1st, 2nd and 4th weeks, it was lower than that of the comparative example 1 and the control group. The statistical results showed that at the 2nd and 4th weeks, the MI value of the experimental group was significantly different from that before the sample was applied (p < 0.05), and the MI value of the experimental group was also significantly reduced compared with the control group.

[0112] Table 2: Skin MI value test (mean ± SD, n = 3)

[0113] Detection temperature Week 0 Week 1 Week 2 Week 4 Control group 230 229 231 230 Example 1 230 220 201 173 Example 2 230 219 196 174 Example 3 230 220 199 170 Example 4 230 218 195 168 Comparative Example 1 230 220 210 199 Comparative Example 2 230 223 208 194

[0114] Skin L* Value

[0115] The test results of the skin L* value are shown in Table 3 below. As can be seen from Table 3, within 4 weeks of the test, after the subjects used the propolis liposome gel prepared according to Examples 1 to 4 of the present application, the L* value showed an upward trend, and at the 1st, 2nd and 4th weeks, it was higher than that of the comparative example 1 and the control group. The statistical results show that at the 2nd and 4th weeks, the L* value of the experimental group was significantly different from that before the sample was applied (p < 0.05); the L* value of the experimental group was significantly different from that of the control group (p < 0.05), indicating that the propolis liposome gel of the present application can effectively improve the color of the skin.

[0116] Table 3: Skin L* value test (mean ± SD, n = 3)

[0117] Detection temperature Week 0 Week 1 Week 2 Week 4 Control group 60 60 60 60 Example 1 60 63 65 67 Example 2 60 63 66 68 Example 3 60 62 65 67 Example 4 60 63 66 68 Comparative Example 1 60 61 62 62 Comparative Example 2 60 61 61 62

[0118] Skin ITA° value

[0119] The test results of the skin ITA° value are shown in Table 4 below. As can be seen from Table 4, within the 4 weeks of the test, after the subjects used the propolis liposome gel prepared according to Examples 1 to 4 of the present application, the ITA° value showed an upward trend, and at the 1st, 2nd and 4th weeks, it was higher than that of the comparative example 1 and the control group. The statistical results showed that at the 2nd and 4th weeks, the ITA° value of the experimental group was significantly different from that before the sample was applied (p < 0.05), indicating that the propolis liposome gel of the present application can significantly improve the dullness of the skin and improve the brightness of the skin.

[0120] Table 4: Skin ITA° value test (mean ± SD, n = 3)

[0121]

[0122]

[0123] VISIA effect analysis before and after using the propolis liposome gel according to the present application

[0124] Before and after 4 weeks of using the propolis liposome gel according to the present application, full-face photos of the subjects were taken using the VISIA facial image analysis system. The results showed that before using the lotion, the subjects' skin was dark yellow with scattered small spots; after 4 weeks of use, the skin texture became finer, the color became whiter, and the spots became lighter. Compared with before and after use, the number of spots decreased by 18%, and the brown spots decreased by 8%, indicating that the propolis liposome gel according to the present application also has a certain spot-lightening effect and effectively reduces the deposition of melanin.

[0125] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A propolis liposome, wherein the propolis liposome is prepared according to the following method: Weigh propolis, crush it, add 4-5 times the volume of co-solvent to extract for 12-36 hours, filter to obtain propolis liquid, add dispersion medium to the propolis liquid, continue to react for 0.5 to 2 hours at 60-85° C. and 500-600 r / min under stirring conditions to obtain propolis liposomes; Preferably, the co-solvent is an ethanol solution with a volume percentage concentration of 70 to 95%; Preferably, the dispersion medium consists of phosphatidylethanolamine (cephalin PE), weakly acidic anionic surfactant disodium lauryl sulfosuccinate and water; Preferably, the mass ratio of the propolis liquid to the dispersion medium is 0.5:1 to 1.5:1, more preferably 0.8:1 to 1.2:1, and most preferably 1:1; Preferably, the mass ratio of phosphatidylethanolamine, disodium lauryl sulfosuccinate and water is (4-6):(0.01-0.8):1; preferably (2.4-3):(0.04-0.5):1, and most preferably 2.4:0.08:1; Preferably, the reaction temperature is 65 to 80°C, most preferably 70-75°C; Preferably, the reaction time is 0.5 to 1.5 hours, most preferably 1 hour; Preferably, the propolis liposomes according to the present invention do not contain glycerol components.

2. A propolis gel comprising the following active ingredients in parts by weight: 0.1-10 parts of the propolis liposome according to claim 1, 2-25 parts of Glycyrrhiza glabra root extract, 1-25 parts of Aloe vera leaf extract, 0.5-20 parts of Centella asiatica extract, 0.5-20 parts of Snow Lotus extract, 0.2-20 parts of Paeonia lactiflora root extract and 0.1-20 parts of Portulaca oleracea extract, and gel-acceptable excipients; The acceptable excipients of the gel include 0.1-10 parts of gel matrix carbomer, 0.1-10 parts of pH value regulator, 0.05-2 parts of surfactant, and 20-75 parts of cosolvent; Preferably, the gel matrix is ​​carbomer; Preferably, the surfactant is Tween 80; Preferably, the pH regulator is triethanolamine; Preferably, the co-solvent is ethanol with a volume percentage concentration of 70 to 95%.

3. The propolis gel according to claim 2, characterized in that The propolis gel comprises the following active ingredients in parts by weight: 0.2-5 parts of the propolis liposome according to claim 1, 5-20 parts of Glycyrrhiza glabra root extract, 2-20 parts of Aloe barbadensis leaf extract, 1-15 parts of Centella asiatica extract, 1-15 parts of Snow Lotus extract, 0.5-15 parts of Paeonia lactiflora root extract and 0.5-15 parts of Portulaca oleracea extract; Preferably, the acceptable excipients of the gel include 0.2-5 parts of gel matrix, 0.2-5 parts of pH regulator, 0.05-1 parts of surfactant, and 40-60 parts of cosolvent.

4. The propolis gel according to claim 2, characterized in that The propolis gel comprises the following active ingredients in parts by weight: 0.3-2.5 parts of the propolis liposome according to claim 1, 6-15 parts of Glycyrrhiza glabra root extract, 5-11 parts of Aloe barbadensis leaf extract, 5-8 parts of Centella asiatica extract, 4-9 parts of Snow Lotus extract, 2-9 parts of Paeonia lactiflora root extract and 3-7 parts of Portulaca oleracea extract; Preferably, the acceptable excipients of the gel include 0.8-3 parts of carbomer, 0.8-2 parts of triethanolamine, 0.3-1.5 parts of Tween 80, and 7-13 parts of ethanol with a volume percentage concentration of 70-95%.

5. The propolis gel according to claim 2, characterized in that The propolis gel comprises the following active ingredients in parts by weight: 2.2-2.7 g of the propolis liposome according to claim 1, 13-17 g of Glycyrrhiza glabra root extract, 8-10 g of Aloe vera leaf extract, 6-8 g of Centella asiatica extract, 4-6 g of Snow Lotus extract, 4-6 g of Paeonia lactiflora root extract and 3-5 g of Portulaca oleracea extract; Preferably, the acceptable excipients of the gel include 2.8-3.2 g of carbomer, 1.1-1.3 g of triethanolamine, 0.2-0.4 g of Tween 80, and 8-12 g of ethanol with a volume percentage concentration of 90%.

6. The method for preparing propolis gel according to claim 2, comprising the following steps: (1) Preparation of plant extracts: Glycyrrhiza glabra root, Aloe vera leaf, Centella asiatica, Saussurea involucrata, Paeonia lactiflora root and Portulaca oleracea are mixed or extracted independently by percolation method, and concentrated to a relative density of 0.8-1.5 mg / ml at 60° C. to obtain respective plant extracts; (2) Preparation of gel forming agent: mixing the gel matrix and part of the co-solvent, and fully swelling the mixture to obtain the gel forming agent; (3) Add propolis liposomes and plant extracts to the gel forming agent, mix well, add the remaining surfactant and pH regulator in sequence, and then add the remaining cosolvent and purified water to a sufficient amount to obtain propolis liposome gel.

7. The method for preparing propolis gel according to claim 6, characterized in that: In the percolation extraction process described in step (1), the percolation solvent is 70-95% ethanol by volume, the mass ratio of the percolation solvent to the plant raw material is 1-20, and the percolation solvent flow rate is 1-15 ml / min; More preferably, the diafiltration solvent is 80-95% ethanol, the volume mass ratio of the diafiltration solvent to the plant material is 1-15, and the diafiltration solvent flow rate is 2-10 ml / min.

8. Use of the propolis gel according to claim 2 in the preparation of skin care products, wherein the skin care products include lotions, emulsions, cosmetic creams, facial masks, paste gels, sprays, facial cleansers, foundations, powder creams, body lotions or massage creams, etc.

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