HK1255152B - Liposome, liposome liquid and cosmetic, and method for producing the liposome - Google Patents

Description

Liposomes, liposome solutions and cosmetics, and methods for manufacturing the above-mentioned liposomes.

Technical Field

This invention relates to liposomes containing proteoglycans in their membranes, liposome solutions containing the above-mentioned liposomes, cosmetics, and methods for manufacturing the above-mentioned liposomes.

Background Technology

The skin, from its outermost layer, comprises the epidermis, dermis, and subcutaneous tissue, and functions as a barrier to protect the organism from external stimuli. The dermis and epidermis are composed of epidermal cells, fibroblasts, and the extracellular matrix of dermal cells. Proteoglycans, along with collagen and other proteins, are the main biomolecules in the extracellular matrix of connective tissue. Young skin maintains its moisture, softness, and elasticity through the constant interaction of these skin tissues, resulting in a firm and radiant appearance.

The outermost layer of the epidermis is the stratum corneum. The stratum corneum is the last layer of skin cells to be formed, consisting of 10-20 layers of corneocytes. These corneocytes contain natural moisturizing factors such as amino acids, urea, and lactic acid. These substances bind with water to retain moisture and keep the skin supple. The outermost layer of the stratum corneum contains more anucleated dead cells, but its barrier function—including retaining moisture and preventing dryness—is crucial to the skin's appearance.

However, the moisture content of the stratum corneum decreases due to external factors such as dryness and ultraviolet radiation, as well as internal factors such as aging, visceral diseases, and stress. This leads to a weakened skin barrier function, causing rough skin, dry skin, and various types of dermatitis. Therefore, the development of moisturizers to maintain the moisture content of the stratum corneum is essential in the fields of cosmetics and dermatology.

On the other hand, in the cosmetics field, liposomes are frequently used to encapsulate active ingredients with moisturizing, antioxidant, and whitening effects on the skin. Liposomes are lipid bilayers or multilayers mainly composed of phospholipids, possessing a structure similar to a cell membrane. Regarding the encapsulation of active ingredients, in the case of water-soluble drugs, they are mostly absorbed into the aqueous phase of the liposome.

Existing technical documents

Patent documents

Patent Document 1: Japanese Patent Application Publication No. 2012-236808

Summary of the Invention

The problem the invention aims to solve

For example, Patent Document 1 describes that: by using cosmetics containing cell growth factors, skin cell proliferation and generation can be effectively promoted, and skin can be improved to have a firm and youthful appearance. It also describes that: from the viewpoint of moderately inhibiting water evaporation and preventing the function of cell growth factors from being inhibited, proteoglycans are encapsulated in the aqueous phase of lipid bodies.

However, we expect further improvements that balance increased skin hydration and improved skin firmness.

The present invention was made in view of the above circumstances, and its object is to provide a liposome that takes into account both the increase of skin moisture and the improvement of skin firmness, a liposome fluid containing the above liposome and a cosmetic, and a method for manufacturing the above liposome.

Solution for solving the problem

The inventors discovered that by using liposomes containing proteoglycans in their membranes (liposome layers), the firmness of the skin is significantly improved, thus completing this invention. That is, this invention is as follows.

(1) A liposome, the membrane of which contains proteoglycans.

(2) A liposome fluid containing the liposomes described in (1) above.

(3) A cosmetic product containing the liposomes described in (1) above.

(4) The use of the liposomes described in (1) above or the liposome fluid described in (2) above, for inclusion in cosmetics.

(5) A method for manufacturing liposomes, comprising a method for manufacturing liposomes whose membranes contain proteoglycans.

The aforementioned liposomes are manufactured by contacting a thin film formed by removing the solvent from a solution in which proteoglycans and lipids are dissolved or dispersed in a solvent with an aqueous solution or water.

The effects of the invention

According to the present invention, liposomes that simultaneously increase skin moisture and improve skin firmness, liposome solutions containing the above-mentioned liposomes, cosmetics, and methods for manufacturing the above-mentioned liposomes can be provided.

Detailed Implementation

<Liposomes whose membranes contain proteoglycans>

In the liposomes of the present invention, the membrane (liposome layer) of the liposome contains proteoglycans. These proteoglycans can be either a membrane component of the liposome or can be ingested into the intercellular space of the liposome. Preferably, the proteoglycans exist in the form of a membrane component of the liposome.

As a liposome membrane containing proteoglycans, it can be a single-layer lipid bilayer or a multilayer lipid bilayer containing multiple layers.

It should be noted that the aqueous phase of the liposomes of the present invention can be simple water, or it can contain or not contain proteoglycans, and it can also contain other physiologically active ingredients.

The source of the proteoglycan in this invention is not particularly limited, and examples include proteoglycans derived from fish cartilage. The type of fish is not particularly limited, but examples include trout (Oncorhynchus gorbuscha, Oncorhynchus masou, Oncorhynchus masouishikawae, etc.), salmon (chum salmon, sockeye salmon, coho salmon, large-scaled salmon, hardhead trout, etc.), sharks, and cod. Fish from the genus Salmon in the family Salmonidae are preferred, with salmon or trout being particularly preferred. Furthermore, the type of cartilage is not particularly limited, but neck cartilage is preferred, with nasal cartilage being the most preferred. In addition, since the neck of fish is usually discarded when processed into food products, neck cartilage has the advantages of low cost and a stable, large-volume supply.

The content of proteoglycans in the liposomes of the present invention is not particularly limited within the range that does not impair the effect of the present invention, and is preferably 0.1 to 30% by mass relative to the total mass of the liposome membrane components, more preferably 0.5 to 20% by mass, further preferably 1 to 15% by mass, and particularly preferably 3 to 10% by mass.

The lipid membrane components constituting the liposomes of the present invention contain at least phospholipids, glycolipids, sterols, glycols, cationic lipids, and polyethylene glycol (PEG) modified lipids (e.g., PEG-phospholipids). Phospholipids and/or glycolipids are preferred.

As usable phospholipids, there are no particular restrictions on the use of, for example, phosphatidylcholine (lecithin), phosphatidylserine, phosphatidic acid, phosphatidylglycerol, dioleoylphosphatidylcholine, distearylphosphatidylcholine, dipalmitoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, sphingomyelin, diceryl phosphate, lysophosphatidylcholine (lysophosphatidylcholine), egg yolk lecithin, soybean lecithin, or hydrogenated phospholipids thereof.

The above-mentioned phospholipids can be used in combination of one or more types.

Examples of glycolipids include: digalactosylglycerol, galactosylglycerol sulfate and other glycerides, galactosylceramide, galactosylceramide sulfate, lactosylceramide, ganglioside G7, ganglioside G6, ganglioside G4 and other glycosphingolipids.

In addition to the lipids mentioned above, other substances can be added as needed to form the liposome membrane. Examples include sterols that act as lipid membrane stabilizers, such as cholesterol, dihydrocholesterol, cholesterol esters, phytosterols, sitosterol, stigmasterol, campesterol, cholesterolanol, or lanosterol. Furthermore, sterol derivatives such as 1-O-sterol glucoside, 1-O-sterol maltodextrin, or 1-O-sterol galactoside are also known to have liposome-stabilizing effects. Cholesterol is particularly preferred.

As for the amount of sterols used, the molar ratio of phospholipids (excluding PEG-phospholipids) to sterols is preferably 100/60 to 100/90, more preferably 100/70 to 100/85. This molar ratio is based on the amount of phospholipids other than PEG-phospholipids. When the molar ratio is 100/60 to 100/90, the stabilization produced by the sterols can be fully utilized to improve the dispersibility of the mixed lipids of phospholipids (excluding PEG-phospholipids) and sterols.

In addition to the aforementioned sterols, diols can also be added as components of the liposome membrane. When diols are added together with phospholipids during liposome fabrication, the retention efficiency of water-soluble compounds within the liposome is improved. Examples of diols include ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, trimethylene glycol, and 1,4-butanediol. The amount of diol used is preferably 0.01 to 20% by mass relative to the total mass of the lipids, more preferably 0.5 to 10% by mass.

The particle size of the liposomes of the present invention is not particularly limited without impairing the effects of the present invention. Particle sizes ranging from 10 nm to 100 μm are examples. From the viewpoint of more effectively achieving the effects of the present invention, a particle size of 50 nm to 50 μm is preferred, and a particle size of 100 nm to 10 μm is more preferred. The particle size of the liposomes can be determined, for example, by a Coulter particle counter (Beckman Coulter Corporation: Submicron Particle Size Analyzer N5).

<Methods for manufacturing liposomes>

As for the manufacturing method of the above-mentioned liposomes, there are no particular limitations without impairing the effect of the present invention. Examples include: (1) a method of manufacturing the above-mentioned liposomes by contacting an aqueous solution or water with a thin film formed by removing the aforementioned organic solvents from a mixture of proteoglycans and lipids and organic solvents (chloroform, methanol, etc.) by vacuum removal or the like (so-called Bangham method) (Japanese Patent Application Publication No. 2011-032230); (2) a method of preparing liposomes by mixing proteoglycans and lipids with organic solvents such as ether or ethanol, injecting the resulting mixture into an aqueous solution such as a buffer solution or water, and then removing the organic solvents; and further, a mechanized method. Methods include (3) ultrasonic treatment (Japanese Patent Application Publication No. 2005-41869); (4) high-pressure homogenizer and high-speed rotary disperser (Japanese Patent Application Publication No. 11-139961); (5) high-pressure filtration using a polycarbonate membrane filter; (6) preparing an oil phase containing proteoglycans and lipids in the upper part of the aqueous phase, introducing droplets into the oil phase, and using centrifugal force to carry the droplets into the aqueous phase; (7) mixing proteoglycans and lipids with an organic solvent miscible with water, such as ethanol, and then replacing the organic solvent with water using a dialysis membrane (Japanese Patent Application Publication No. 01-224042), etc.

As a particularly preferred method, a method of manufacturing liposomes is preferred by contacting a thin film formed by removing the aforementioned solvent from a solution in which proteoglycans and lipids are dissolved or dispersed in the solution with an aqueous solution or water.

Examples of solvents include water, ethanol, lower alcohols such as butanol, and organic solvents such as chloroform. In particular, proteoglycans and lipids are difficult to dissolve when water is used; therefore, it is preferable to disperse them using methods such as ultrasound to form a solution. When forming a thin film from this solution, drying can be performed while the solution is in contact with the substrate. For example, the aforementioned solution can be added to a glass flask, and the solvent can be removed using a rotary evaporator while heating, forming a thin film of proteoglycans and lipids on the inner wall (substrate) of the flask.

In addition, as a method for bringing a thin film into contact with an aqueous solution or water, methods such as forming a thin film on the wall (substrate) of any container and then oscillating (swelling) the aforementioned thin film in the presence of an aqueous solution or water, such as a buffer solution, can be listed.

There are no particular restrictions on the type of container used to form the vessel wall; examples include flasks, etc.

Furthermore, if necessary, the film can be further peeled off from the aforementioned container wall using mechanical stirring to produce the aforementioned liposomes.

<Lipobody Fluid>

The liposome fluid of the present invention is a liquid containing the above-mentioned liposomes.

The liposome fluid of the present invention can be a liquid in which the separated liposomes are added to any liquid, or it can be a liquid containing liposomes prepared by the above-described method for manufacturing liposomes.

Furthermore, the liposome fluid of the present invention may be appropriately combined with other components, such as anionic surfactants, amphoteric surfactants, nonionic surfactants, viscosity modifiers, oils, powders (pigments, resins, pigments, etc.), preservatives such as phenoxyethanol, fragrances, humectants such as 1,3-butanediol, physiologically active ingredients, salts, solvents, antioxidants, chelating agents, pearlizing agents, neutralizing agents, pH adjusters, enzymes, etc. Specific examples of the combined components are shown below, but are not limited to these.

As anionic surfactants, examples include α-acylsulfonates, alkylsulfonates, alkylallyl sulfonates, alkylnaphthalene sulfonates, alkyl sulfates, alkyl ether sulfates, alkylamide sulfates, polyoxyethylene alkyl ether sulfates, polyoxyethylene alkylamide ether sulfates, alkyl phosphates, alkylamide phosphates, alkylacylalkyl taurines, N-acyl amino acid salts, sulfosuccinates, and perfluoroalkyl phosphates.

In addition, examples of amphoteric surfactants include glycine-type, aminopropionic acid-type, carboxybetaine-type, sulfobetaine-type, sulfonic acid-type, sulfuric acid-type, and phosphoric acid-type surfactants. Preferred examples include 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazoline betaine and coconut oil fatty acid amamidopropyl betaine. Furthermore, in addition to the purpose of forming liposomes, phospholipids such as lecithin can also be used as amphoteric surfactants.

Examples of nonionic surfactants include fatty acid alkanolamides, polyoxyethylene hydrogenated castor oil, polyoxyethylene dehydrated sorbitan fatty acid esters, polyoxyethylene alkyl ethers, polyoxyethylene alkyl esters, sucrose fatty acid esters, polyglycerol fatty acid esters, and alkylamine oxides.

Examples of viscosity modifiers include: acrylic amides and their derivatives, carboxyvinyl polymers, alkyl-modified carboxyvinyl polymers, cellulose, keratin and collagen or their derivatives, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, high-methoxyl pectin, low-methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabinogalactan, ebony gum, tragacanth gum, alginate, albumin, casein, gel polysaccharides, gellan gum, dextran, etc.

As oil-based agents, examples include volatile and non-volatile oils, solvents, and resins commonly used in cosmetics, which can be liquid, paste, or solid at room temperature. Examples of oil-based agents include: higher alcohols such as cetyl alcohol, isostearyl alcohol, lauryl alcohol, hexadecyl alcohol, and octyldodecyl alcohol; fatty acids such as isostearic acid, undecenoic acid, and oleic acid; and myristyl myristate, hexyl laurate, octyldodecyl myristate, decyl oleate, isopropyl myristate, hexadecyl dimethyl octanoate, glyceryl monostearate, glyceryl tricaprylate, diethyl phthalate, ethylene glycol monostearate, and hydroxyl groups. Esters such as octyl stearate, cholesterol stearate, cholesterol oleate, and cholesterol branched-chain fatty acid esters; hydrocarbons such as liquid paraffin, petrolatum, and squalane; waxes such as lanolin, reduced lanolin, and carnauba wax; oils such as mink oil, cocoa butter, coconut oil, palm kernel oil, camellia oil, sesame oil, castor oil, and olive oil; and silicone oils such as dimethyl polysiloxane, cyclic dimethyl polysiloxane, and methylphenyl polysiloxane.

As powders, examples include: pigments such as Red 201, Yellow 4, Blue 1, and Black 401; lake pigments such as Yellow 4 aluminum lake and Yellow 203 Ba lake; resins such as nylon powder, silk powder, organosilicon powder, cellulose powder, organosilicon elastomer spherical powder, and polyethylene powder; colored pigments such as yellow iron oxide, red iron oxide, chromium oxide, carbon black, ultramarine, and Prussian blue; white pigments such as zinc oxide and titanium dioxide; extender pigments such as talc, mica, sericite, and kaolin; pearlescent pigments such as mica titanium; metal salts such as barium sulfate, calcium carbonate, magnesium carbonate, and magnesium silicate; inorganic powders such as silicon dioxide and aluminum oxide; bentonite; montmorillonite; and boron nitride. There are no particular restrictions on the shape of these powders (spherical, rod-shaped, needle-shaped, plate-shaped, irregular, scaly, spindle-shaped, etc.).

As physiologically active ingredients, substances that provide any physiological activity to the skin when applied can be listed. Examples include: anti-aging agents, UV protectants, skin tightening agents, antioxidants, moisturizers, blood circulation promoters, antibacterial agents, bactericides, desiccants, cooling agents, warming agents, vitamins, amino acids, wound healing promoters, irritant soothing agents, analgesics, cell activators, and enzyme components.

<Cosmetics>

The cosmetic of the present invention contains the above-mentioned liposomes.

As a cosmetic product of the present invention, the above-mentioned liposome can be used as is, but it is preferred to use the above-mentioned liposome containing the cosmetic product.

As cosmetics, there are no restrictions on their classification. They broadly include various cosmetics and cosmetic products, such as basic cosmetics like toners, lotions, serums, masks, scrubs, and facial cleansers; color cosmetics like lipsticks and foundations; cosmetic products like body soaps, soaps, shampoos, and conditioners; and hair-care products like hair treatments.

Example

The present invention is illustrated in detail below through examples, but these examples do not constitute a limitation on the scope of the present invention.

Using the raw materials with the compositions shown in Table 1 below, liposomes 1 to 4 were prepared by the following preparation method.

Table 1

Lipobody fluid 1 Lipofluid 2 Lipofluid 3 Lipofluid 4 Lecithin content 1 1 1 1 Proteoglycan content 0.05 0.05 - 0.05 Proteoglycan position membrane Encapsulation - extracellular Purified water content margin margin margin margin

[Unit: mass %]

• Proteoglycan: derived from salmon nasal cartilage (manufactured by Wako Pure Chemical Industries, Ltd.)

(1) Lipofluid 1

The liposome membrane components (lecithin and proteoglycan) were mixed in a water/ethanol mixture (1/9 by volume), and the solvent was removed using an evaporator to prepare a membrane. Purified water was added, and the membrane was peeled off by shaking to prepare a liposome fluid containing proteoglycan.

(2) Lipobody fluid 2

The liposome membrane component (lecithin) was mixed into a water/ethanol mixture (1/9 by volume), and the solvent was removed using an evaporator to prepare a membrane. An aqueous solution of proteoglycan dissolved in purified water was added to the membrane, and the membrane was peeled off by shaking to prepare a proteoglycan-encapsulated liposome solution.

(3) Lipofluid 3

The liposome membrane component (lecithin) was mixed into a water/ethanol mixture (1/9 by volume), and the solvent was removed using an evaporator to prepare a membrane. Purified water was added, and the membrane was peeled off by shaking to prepare a proteoglycan-free liposome solution 3.

(4) Lipofluid 4

The liposome membrane component (lecithin) was mixed in a water/ethanol mixture (1/9 by volume), and the solvent was removed using an evaporator to prepare a membrane. Purified water was added, and the membrane was peeled off by shaking to prepare liposomes. Proteoglycans were dissolved in the mixture to prepare a liposome fluid containing proteoglycans outside the liposome membrane.

<Examples and Comparative Examples 1-4>

Using the above-mentioned liposomes 1 to 4, the cosmetic lotions of the examples and comparative examples 1 to 4 with the compositions shown in Table 2 below were prepared according to conventional methods.

Table 2

[Unit: mass %]

The lotions from Examples 1 to 4 and Comparative Examples 1 to 4 were evaluated.

<Evaluation Methods>

Ten inspectors were randomly selected, and the coating area was set on the inside of their forearms. An appropriate amount of the test sample was applied twice a day, morning and evening. In addition, the usage period was set at 1 week.

The corneal moisture content of the uncoated and coated evaluation samples was determined using a SKICON-200EX (I.B.S. manufactured) corneal moisture meter one week after coating. The increase or decrease in corneal moisture content after coating was calculated when the corneal moisture content before coating was set to 100.

In addition, a questionnaire survey was conducted to determine whether the "skin firmness" had improved, using the following criteria.

Improvement: 1 point; No improvement: 0 points. Evaluation is based on the sum of the scores.

The evaluation results are shown in Table 3.

Table 3

According to the results shown in Table 3, the toner containing proteoglycans in the liposome membrane of the example achieved both increased skin hydration and improved skin firmness compared to Comparative Example 2, which used liposome fluid 3 without proteoglycans, Comparative Example 3, which used liposome fluid 4 with proteoglycans present outside the liposome membrane, and Comparative Example 4, which did not contain proteoglycans or liposomes.

It is evident that, in particular, the lotion of Example 1, which contains proteoglycans in the liposome membrane, compared with the lotion of Comparative Example 1, which encapsulates proteoglycans in liposomes, achieves both an increase in skin moisture and an improvement in skin firmness.

Claims (7)

1. A liposome, wherein the membrane of the liposome contains proteoglycans in the form of membrane components of the liposome. 2. The liposome according to claim 1, wherein the content of the proteoglycan is 0.1 to 30% by mass relative to the total mass of the liposome membrane components. 3. A liposome fluid comprising the liposomes as described in claim 1 or 2. 4. A cosmetic product comprising the liposomes as described in claim 1 or 2. 5. Use of the liposomes of claim 1 or the liposome fluid of claim 3, for inclusion in cosmetics. 6. A method for manufacturing liposomes, comprising the method of manufacturing liposome membranes containing proteoglycans in the form of liposome membrane components. The liposomes are manufactured by contacting a thin film formed by removing the solvent from a solution in which proteoglycans and lipids are dissolved or dispersed in a solvent with an aqueous solution or water. 7. The method according to claim 6, wherein the content of the proteoglycan is 0.1 to 30% by mass relative to the total mass of the liposome membrane components.