JP6883723B2 - Liposomes, liposome solutions and cosmetics, and methods for producing the above liposomes. - Google Patents

Description

The present invention relates to liposomes in which the membrane of the liposome contains proteoglycan, a liposome solution and cosmetic containing the liposome, and a method for producing the liposome.

The skin consists of the epidermis, dermis, and subcutaneous tissue from the outside, and its barrier function protects the living body from external stimuli. The dermis and epidermis of the skin are composed of epidermal cells, fibroblasts, dermis extracellular matrix and the like. Proteoglycan is a major biopolymer that forms a substrate in the extracellular matrix of connective tissue together with collagen and the like. In young skin, the interaction of these skin tissues maintains homeostasis, thereby ensuring water retention, flexibility, elasticity, etc., and maintaining a state of firmness and luster in appearance.

The outermost layer of the epidermis further has a stratum corneum. The stratum corneum is the part where cells consisting of 10 to 20 layers of corneocytes finally reach. Natural moisturizing factors such as amino acids, urea, and lactic acid are present in corneocytes, and when they combine with water, they retain water and maintain the flexibility of the skin. In the stratum corneum, dead cells without nuclei can be seen toward the outside, but the barrier function of the stratum corneum, such as the function of retaining water and preventing dryness, is greatly related to the beauty of the skin.
However, the water content of the stratum corneum is reduced by external factors such as dryness and ultraviolet rays and internal factors such as aging, aging, visceral diseases, and stress, resulting in a decrease in the barrier function of the skin, resulting in rough skin and dry skin. It is said to cause various dermatitis. Therefore, in the fields of cosmetics and dermatology, it is indispensable to develop a moisturizer that maintains the water content of the stratum corneum.

On the other hand, in the field of cosmetics, it is common practice to enclose an active ingredient in a liposome as a means for delivering an active ingredient having a moisturizing function, an antioxidant effect, a whitening effect, and the like to the skin. Liposomes are lipid bilayers or multiple layers mainly composed of phospholipids, and have a structure similar to a cell membrane. Encapsulation of the active ingredient is often incorporated into the internal aqueous phase of the liposome in the case of water-soluble drugs.

Japanese Unexamined Patent Publication No. 2012-236808

For example, according to Patent Document 1, according to a cosmetic containing a cell growth factor, skin cells can be efficiently proliferated and generations are changed, and the skin can be improved to be firm and youthful. It is stated that it is suggested that proteoglycan is encapsulated in the aqueous phase of liposomes from the viewpoint of preventing the function of cell growth factors and the like from being inhibited while appropriately suppressing the evaporation of water. ing.
However, it has been desired to further improve both the increase in skin moisture content and the improvement of skin firmness.

The present invention has been made in view of the above circumstances, and provides a liposome that achieves both an increase in skin moisture content and an improvement in skin firmness, a liposome solution and cosmetic containing the liposome, and a method for producing the liposome. The purpose.

The present inventor has found that the firmness of the skin is particularly improved by using a liposome having a liposome membrane (lipid layer) containing proteoglycan, and has completed the present invention. That is, the present invention is as follows.

(1) Liposomes in which the membrane of the liposome contains proteoglycan.
(2) A liposome solution containing the liposome according to (1) above.
(3) A cosmetic containing the liposome according to (1) above.
(4) A method for producing a liposome whose liposome membrane contains proteoglycan.
A method for producing the liposome by contacting a thin film formed by removing the solvent from a solution in which proteoglycan and a lipid are dissolved or dispersed in a solvent with an aqueous solution or water.

According to the present invention, it is possible to provide a liposome that achieves both an increase in skin moisture content and an improvement in skin firmness, a liposome solution containing the liposome, a cosmetic, and a method for producing the liposome.

<Liposome membrane containing proteoglycan, liposome>
In the liposome of the present invention, the membrane (lipid layer) of the liposome contains proteoglycan. The proteoglycan may be a membrane component of liposomes or may be incorporated between the membranes of liposomes. Above all, it is preferable that proteoglycan is present as a membrane component of liposomes.
The liposome membrane containing proteoglycan may be a single-layer lipid bilayer or a multi-layer composed of a plurality of lipid bilayers.
The internal aqueous phase of the liposome of the present invention may be simple water, may or may not contain proteoglycan, and may contain other physiologically active ingredients.

The origin of proteoglycan in the present invention is not particularly limited, and examples thereof include proteoglycan derived from fish cartilage. The fish are not particularly limited, and specific examples thereof include trout (pink salmon, cherry salmon, satsukimasu, etc.), salmon (white salmon, sockeye salmon, coho salmon, chinook salmon, steel head, etc.), sharks, cod, and the like. Fish of the genus Salmon in the family Salmonidae are preferred, especially salmon or trout. The cartilage is not particularly limited, but head cartilage, particularly nasal cartilage, is preferable. In addition, since the head is usually discarded when fish are processed into food products, the cost of obtaining head cartilage is low, and there is an advantage that a large amount of cartilage can be stably supplied.

The content of proteoglycan in the liposome of the present invention is not particularly limited as long as the effect of the present invention is not impaired, but is preferably 0.1 to 30% by mass with respect to the total mass of the liposome membrane component. It is more preferably 5 to 20% by mass, further preferably 1 to 15% by mass, and particularly preferably 3 to 10% by mass.

The lipid membrane component constituting the lipid membrane of the liposome of the present invention includes at least phospholipids, glycolipids, sterols, glycols, cationic lipids, polyethylene glycol (PEG) -modified lipids (for example, PEG-phospholipids) and the like. Is done. Of these, phospholipids and / or glycolipids are preferably used.

Examples of phospholipids that can be used include phosphatidylcholine (lecithin), phosphatidylserine, phosphatidylic acid, phosphatidylglycerin, dioleoylphosphatidylcholine, distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, palmitoyloleoil phosphatidylcholine, phosphatidylethanolamine, and phosphatidylethanolamine. , Disetyl phosphate, lysophosphatidylcholine (lysolecithin), egg yolk lecithin, soybean lecithin, hydrogenated phospholipids thereof and the like can be used without particular limitation.
The above phospholipids may be used alone or in combination of two or more.

Examples of the glycolipid include glycerolipids such as digalactosyl diglyceride and galactosyl diglyceride sulfate ester, and sphingolipids such as galactosylceramide, galactosylceramide sulfate, lactosylceramide, ganglioside G7, ganglioside G6 and ganglioside G4.

In addition to the above lipids, other substances may be added as constituents of the liposome membrane, if necessary. For example, sterols acting as a lipid membrane stabilizer, such as cholesterol, dihydrocholesterol, cholesterol ester, phytosterol, sitosterol, stigmasterol, campesterol, cholestanol, lanosterol and the like can be mentioned. Further, sterol derivatives such as 1-O-sterol glucoside, 1-O-sterol maltoside or 1-O-sterol galactoside are also known to be effective in stabilizing liposomes. Of these, cholesterol is particularly preferred.

As the amount of sterols used, the molar ratio of phospholipids (without PEG-phospholipids) / sterols is preferably 100/60 to 100/90, and more preferably 100/70 to 100/85. preferable. This molar ratio is based on the amount of phospholipids excluding PEG-phospholipids. When the molar ratio is 100/60 to 100/90, stabilization by sterols that improves the dispersibility of mixed lipids of phospholipids (excluding PEG-phospholipids) and sterols can be sufficiently exhibited.

In addition to the above sterols, glycols may be added as a constituent component of the liposome membrane. When glycols are added together with phospholipids and the like when producing liposomes, the retention efficiency of the water-soluble compound in the liposomes increases. Examples of glycols include ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, trimethylene glycol, 1,4-butanediol and the like. The amount of glycols used is preferably 0.01 to 20% by mass, more preferably 0.5 to 10% by mass, based on the total mass of lipids.

The particle size of the liposome of the present invention is not particularly limited as long as the effect of the present invention is not impaired, but the particle size is 10 nm to 100 μm, and the particle size is from the viewpoint of more reliably achieving the effect of the present invention. The particle size is preferably 50 nm to 50 μm, and more preferably 100 nm to 10 μm. The particle size of the liposome can be measured by, for example, a Coulter counter (manufactured by Beckman Coulter, Inc .: Submicron Particle Analyzer N5) or the like.

<Liposome manufacturing method>
The method for producing the liposome is not particularly limited as long as the effect of the present invention is not impaired. For example, (1) the organic solvent is depressurized from a mixture of proteoglycan and a lipid and an organic solvent (chloroform, methanol, etc.). A method for producing the liposome by contacting the thin film formed by removing it by removal or the like with an aqueous solution or water (so-called Bangham method) (Japanese Patent Laid-Open No. 2011-032230), (2) Proteoglycan and lipid and ether or ethanol. The method of preparing by mixing with an organic solvent such as, etc., injecting the obtained mixed solution into an aqueous solution such as a buffer solution or water, and then removing the organic solvent, and further, as a mechanical method, (3) Method by sonic treatment (Japanese Patent Laid-Open No. 2005-41869), (4) Method by high-pressure homogenizer or high-speed rotary disperser (Japanese Patent Laid-Open No. 11-139961), (5) Method by high-pressure filtration using a polycarbonate solvent filter , (6) A method in which an oil phase containing a proteoglycan and a lipid is prepared above the aqueous phase, droplets are introduced into the oil phase, and the droplets are dragged into the aqueous phase by the force of centrifugation, (7) Proteoglycan and Examples thereof include a method of mixing a lipid and an organic solvent compatible with water such as ethanol, and then replacing the organic solvent with water via a dialysis membrane (Japanese Patent Laid-Open No. 01-224042).
As a particularly preferable method, a method of producing liposomes by contacting a thin film formed by removing the solvent from a solution in which proteoglycan and a lipid are dissolved or dispersed in a solvent with an aqueous solution or water is preferable.
Examples of the solvent include water, lower alcohols such as ethanol and butanol, and organic solvents such as chloroform. In particular, when water is used, proteoglycans and lipids are difficult to dissolve, so it is preferable to disperse them by ultrasonic waves or the like to prepare a solution. When forming a thin film from this solution, the solution may be brought into contact with the substrate and dried. For example, by putting the solution into a glass flask and using a rotary evaporator while heating, the solvent can be removed and a thin film of proteoglycan and lipid can be formed on the inner wall (substrate) of the flask.
Further, as a method of bringing the thin film into contact with an aqueous solution or water, after forming the thin film on the vessel wall (substrate) of an arbitrary container, the thin film is shaken (swelled) in the presence of an aqueous solution such as a buffer solution or water. The method and the like can be mentioned.
The container forming the vessel wall is not particularly limited, and examples thereof include a flask and the like.
Further, if necessary, the liposome can be produced by further peeling the thin film from the vessel wall by mechanical stirring means.

<Liposome solution>
The liposome solution of the present invention is a solution containing the above-mentioned liposome.
The liposome solution of the present invention may be a solution obtained by adding the isolated liposome to any solution, or may be a solution containing the liposome prepared by the method for producing the liposome as it is. ..
Further, the liposome liquid of the present invention contains other components, for example, anionic surfactant, amphoteric surfactant, nonionic surfactant, viscosity regulator, oil agent, as long as the effect of the present invention is not impaired. Powders (pigments, resins, pigments, etc.), preservatives such as phenoxyethanol, fragrances, moisturizers such as 1,3-butylene glycol, physiologically active ingredients, salts, solvents, antioxidants, chelating agents, pearlizing agents, medium Ingredients such as a Japanese agent, a pH adjuster, and an enzyme can be appropriately blended. Specific examples of the compounding ingredients are shown below, but the present invention is not limited to these.

Anionic surfactants include α-acyl sulfonate, alkyl sulfonate, alkylallyl sulfonate, alkylnaphthalene sulfonate, alkyl sulfate, alkyl ether sulfate, alkylamide sulfate, and polyoxyethylene. Alkyl ether sulfate, polyoxyethylene alkylamide ether sulfate, alkyl phosphate, alkylamide phosphate, alkiloyl alkyl taurine salt, N-acyl amino acid salt, sulfosuccinate, perfluoroalkyl phosphate, etc. Be done.

Examples of the amphoteric tenside include glycine type, aminopropionic acid type, carboxybetaine type, sulfobetaine type, sulfonic acid type, sulfuric acid type, phosphoric acid type and the like, and 2-alkyl-N- is suitable. Examples thereof include carboxymethyl-N-hydroxyethyl imidazolinium betaine and coconut oil fatty acid amide propyl betaine. In addition, phospholipids such as lecithin can be used as an amphoteric surfactant in addition to the purpose of forming liposomes.

Nonionic surfactants include fatty acid alkanolamide, polyoxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene alkyl ester, sucrose fatty acid ester, polyglycerin fatty acid ester, and alkyl. Examples include amine oxides.

Examples of the viscosity modifier include acrylic acid amide and its derivatives, carboxyvinyl polymers, alkyl-modified carboxyvinyl polymers, cellulose, keratin and collagen or its derivatives, calcium alginate, purulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, Examples thereof include high methoxyl pectin, low methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabinogalactan, carrageenan, tragacant gum, alginic acid, albumin, casein, curdlan, gellan gum and dextran.

Examples of the oil agent include volatile and non-volatile oil agents, solvents and resins usually used in cosmetics, and may be liquid, paste, or solid at room temperature. Examples of the oil agent include higher alcohols such as cetyl alcohol, isostearyl alcohol, lauryl alcohol, hexadecyl alcohol, and octyldodecanol, fatty acids such as isostearic acid, undecylenic acid, and oleic acid, myristyl myristate, hexyl laurate, and octyl myristate. Dodecyl, decyl oleate, isopropyl myristate, hexyl decyl dimethyl octanoate, glycerin monostearate, glycerin trioctanoate, diethyl phthalate, ethylene glycol monostearate, octyl oxystearate and other esters, cholesteryl stearate, oleic acid Cholesteryl, cholesterol esters such as branched fatty acid cholesteryl, hydrocarbons such as liquid paraffin, vaseline, squalane, wax such as lanolin, reduced lanolin, carnauba wax, mink oil, cacao butter, palm oil, palm kernel oil, camellia oil, sesame oil, sunflower Examples thereof include oils and fats such as olive oil, and silicone oils such as dimethylpolysiloxane, cyclic dimethylpolysiloxane, and methylphenylpolysiloxane.

As powders, pigments such as Red No. 201, Yellow No. 4, Blue No. 1, Black No. 401, Lake No. 4 Al Lake, Yellow No. 203 Ba Lake and other rake pigments, nylon powder, silk powder, silicone powder, cellulose Powder, silicone elastomer spherical powder, resin such as polyethylene powder, colored pigments such as yellow iron oxide, red iron oxide, chromium oxide, carbon black, ultramarine blue, dark blue, white pigments such as zinc oxide and titanium oxide, talc, mica, Constitution pigments such as sericite and kaolin, pigments of pearl pigments such as mica titanium, metal salts such as barium sulfate, calcium carbonate, magnesium carbonate, magnesium silicate, inorganic powders such as silica and alumina, bentonite, smectite, boron nitride, etc. Can be mentioned. The shape of these powders (spherical, rod-shaped, needle-shaped, plate-shaped, indefinite-shaped, fluff-shaped, spindle-shaped, etc.) is not particularly limited.

Examples of the bioactive component include substances that give some bioactivity to the skin when applied to the skin. For example, anti-aging agents, UV protection agents, tightening agents, antioxidants, moisturizers, blood circulation promoters, antibacterial agents, bactericidal agents, desiccants, cooling agents, warming agents, vitamins, amino acids, wound healing promoters. , Stimulant relievers, analgesics, cell activators, enzyme components and the like.

<Cosmetics>
The cosmetic of the present invention contains the above liposomes.
As the cosmetic of the present invention, the liposome solution can be used as it is, but the liposome solution is suitable for use by being contained in the cosmetic.
There are no restrictions on the category of cosmetics, and they include a wide range of cosmetics, toiletry products, etc., and basic cosmetics such as lotions, emulsions, essences, packs, peeling agents, and wash pigments, lipsticks, and foundations. Examples include make-up cosmetics such as, body soap, soap, shampoo, conditioner, toiletry products such as conditioner, and hair cosmetics such as hair set agents.

Hereinafter, the present invention will be specifically described with reference to Examples, but these do not limit the scope of the present invention.
Liposomal solutions 1 to 4 were prepared by the following preparation methods using each raw material having the composition shown in Table 1 below.

(1) Liposome solution 1
The liposome membrane components (lecithin and proteoglycan) were mixed with a water / ethanol mixed solvent (1/9 (volume ratio)), and then the solvent was removed with an evaporator to prepare a film. Purified water was added thereto, and the film was peeled off by shaking to prepare a liposome solution containing proteoglycan in the membrane.
(2) Liposomal solution 2
The liposome membrane component (lecithin) was mixed with a water / ethanol mixed solvent (1/9 (volume ratio)), and then the solvent was removed with an evaporator to prepare a film. An aqueous solution prepared by dissolving proteoglycan in purified water was added thereto, and the film was peeled off by shaking to prepare a liposome solution containing proteoglycan.
(3) Liposomal solution 3
The liposome membrane component (lecithin) was mixed with a water / ethanol mixed solvent (1/9 (volume ratio)), and then the solvent was removed with an evaporator to prepare a film. Purified water was added thereto, and the film was peeled off by shaking to prepare a liposome solution 3 containing no proteoglycan.
(4) Liposomal solution 4
The liposome membrane component (lecithin) was mixed with a water / ethanol mixed solvent (1/9 (volume ratio)), and then the solvent was removed with an evaporator to prepare a film. Purified water was added thereto, and the film was peeled off by shaking to prepare liposomes. Proteoglycan was dissolved therein to prepare a liposome solution 4 in which proteoglycan was present outside the liposome membrane.

<Examples and Comparative Examples 1 to 4>
Using the above liposome solutions 1 to 4, the cosmetics of Examples and Comparative Examples 1 to 4 having the compositions shown in Table 2 below were prepared by a conventional method.

The lotions of Examples and Comparative Examples 1 to 4 were evaluated.
<Evaluation method>
Ten monitors were arbitrarily extracted, an application site was set on the inside of the forearm, and an appropriate amount of the test sample was applied twice in the morning and evening. The period of use was one week.

The water content of the stratum corneum at the time of non-coating and one week after the coating of each evaluation sample was measured using a stratum corneum moisture meter SKICON-200EX (manufactured by IBS). When the water content of the stratum corneum before coating was set to 100, the rate of increase / decrease in the water content of the stratum corneum after coating was calculated.

At the same time, a questionnaire survey was conducted to see if "skin firmness" had improved, using the following criteria.
Improved: 1 point, not improved: 0 points, and evaluated by the total score.
The evaluation results are shown in Table 3.

As is clear from the results shown in Table 3, the lotion of the example in which the proteoglycan was contained in the liposome membrane was Comparative Example 2 using the liposome solution 3 containing no proteoglycan, and the liposome solution in which the proteoglycan was present outside the liposome membrane. It can be seen that both the increase in skin moisture content and the improvement in skin firmness are achieved as compared with Comparative Example 3 using 4 and Comparative Example 4 containing no proteoglycan and liposome.
In particular, it can be seen that the lotion of Example 1 containing proteoglycan in the liposome membrane achieves both an increase in skin moisture content and an improvement in skin firmness as compared with the lotion of Comparative Example 1 in which the liposome contains proteoglycan.

Claims (4)

Liposomes in which the membrane of the liposome contains proteoglycan derived from fish. A liposome solution containing the liposome according to claim 1. The cosmetic containing the liposome according to claim 1. The liposome membrane is a method for producing liposomes containing fish-derived proteoglycans.
A method for producing the liposome by contacting a thin film formed by removing the solvent from a solution in which proteoglycan and a lipid are dissolved or dispersed in a solvent with an aqueous solution or water.