JP2006249010A - Skin external preparation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical preparation which is excellent in the absorbability or targeting of retinol or its derivative in a living body. <P>SOLUTION: The pharmaceutical preparation, improved in the absorbability of retinol or its derivative into the skin etc., and excellent in sustained release and targeting, is prepared by using the following nanoparticles: primary nanoparticles which contains retinol or its derivative, an organic compound having a hydrophobic group and an anionic group, and a surfactant; secondary nanoparticles which is prepared by causing a bivalent or trivalent metal salt to act on a solution containing the primary nanoparticles; and tertiary nanoparticles which is prepared by further dispersing a monovalent, bivalent, or trivalent basic salt. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a nanoparticle containing retinol or a derivative thereof, and more particularly to a nanoparticle containing retinol or a derivative thereof, a method for producing the same, and an external preparation containing the particle.

  As its name is derived from the retina, retinoid is vitamin A and its analogs that are necessary for the protection of retinal cells, and it is known that deficiency causes symptoms such as night blindness. Furthermore, in recent years, it has been found that it is involved in the control of genetic information and has attracted great interest. Retinoid is a general term for retinol, retinal, retinoic acid and their derivatives. Promyelocytic leukemia, Kaposi's sarcoma, acne, skin photoaging, keratinized skin disease (psoriasis, ichthyosis, etc.), keloid, pigment Various synthetic retinoids have been developed in anticipation of the differentiation-inducing action of retinoids on cancer cells, and are used for the prevention and treatment of deposition and fine wrinkles.

  Furthermore, the action of retinoids on the skin is, for the epidermis, promotion of epithelial cell division, prevention of epidermal layer thickening, promotion of epidermal hyaluronic acid synthesis, induction of epidermal TGF-β expression, stratum corneum peeling, and dermis On the other hand, promotion of dermal collagen production, angiogenesis of the dermal papillary layer, suppression of sebaceous gland function, suppression of MMP and collagen expression, and repair of elastin fiber damage are known.

  In particular, among retinoids, retinol is an endogenous compound that occurs naturally in the living body, and retinol or a derivative thereof is less irritating and less irritating and safer than retinoic acid. Therefore, development of a safe external preparation for skin containing retinol or a derivative thereof is particularly desired among retinoids.

  By the way, retinoids are fat-soluble substances, and external preparations using oily bases such as oil and petrolatum have extremely poor permeability to skin, and retinoic acid and retinol are decomposed by light and heat and are stable. Is known to be bad. For these reasons, in preparations in which retinoids are stably blended, various preparations for preparations are required in order to exert their effects sufficiently.

  So far, preparations aiming at improvement and control of skin permeability and skin retention have been investigated as preparations containing stable retinoids by making particles of retinoids themselves or devising them as a base. . For example, a non-phospholipid liposome composition containing retinoids (Patent Document 1: Japanese Patent Publication No. 11-503165) or retinoic acid is dissolved in a small amount of a highly polar organic solvent and dispersed with water containing an alkali. Then, retinoic acid nanocapsules obtained by adding a divalent metal salt to a mixed micelle obtained by adding a nonionic surfactant and then adding a salt having a divalent anion are known (patent) Document 2: JP-A No. 2004-161739).

However, in the liposome preparation disclosed in Patent Document 1, although the skin permeability of retinoid is high, it is absorbed in the body without staying in the epidermis and dermis, so that it is difficult to exert its effect locally. It is. In addition, in the method using nanocapsules disclosed in Patent Document 2, since retinol or a derivative thereof does not have a free radical, it cannot be solubilized with an alkali, so that the dispersion is not performed sufficiently, and stable production of nanoparticles can be achieved. It is difficult to do.
Therefore, there has been a development of a nanoparticle that stably contains retinol or a derivative thereof, has good absorbability into living bodies including skin and mucous membranes, and can exhibit high bioavailability, and a method for producing these nanoparticles. Expected.
Japanese National Patent Publication No. 11-503165 JP 2004-161739 A

  In view of the above-mentioned present situation, the present invention is excellent in absorbability into a living body including a topical region, and retinol or a derivative thereof is efficiently delivered to an action site by an administration method via skin and mucous membrane. An object is to provide an excellent preparation for external use.

  In order to solve such problems, the present inventors have intensively studied and, as a result, used special nanoparticles that are much smaller than erythrocytes at least partially or completely coated with a divalent or trivalent metal salt, It has been found that it can contain retinol or its derivatives. Furthermore, the present inventors disperse retinol or a derivative thereof, which is a fat-soluble drug and hardly solubilized with alkaline water, in an aqueous medium together with an organic compound having a hydrophobic group and an anion residue, and a surfactant. Thus, it was found that primary nanoparticles can be prepared and used for coating with the metal salt. Such nanoparticles can be prepared to have a desired particle size by adjusting the ratio of each component. As a result of examining administration of the nanoparticles thus obtained with skin and mucosal light oil, the retinol contained in the nanoparticles or a derivative thereof was confirmed to be highly absorbable in vivo, and the present invention was completed. .

  Accordingly, the present invention provides nanoparticles containing retinol or a derivative thereof having excellent transdermal absorbability by dermal or mucosal administration for systemic and topical administration. Moreover, the external preparation containing the said nanoparticle is provided. Furthermore, the nanoparticles of the present invention can be advantageously used for injection.

More specifically, the present invention provides:
(1) Nanoparticles obtained by reacting primary nanoparticles containing retinol or a derivative thereof with a divalent or trivalent metal salt;
(2) Primary nanoparticles containing retinol or a derivative thereof are reacted with a divalent or trivalent metal salt to form secondary nanoparticles, and a monovalent to trivalent basic salt is added to the secondary nanoparticles. Nanoparticles made of acting;
(3) The nanoparticle according to the above 1 or 2, wherein the primary nanoparticle comprises reacting retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant;
I will provide a.

More specifically, the present invention provides:
(4) A divalent or trivalent metal salt in a primary nanoparticle prepared by dispersing retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant in an aqueous medium. Nanoparticles containing retinol or a derivative thereof, wherein
(5) A divalent or trivalent metal salt formed on primary nanoparticles prepared by dispersing retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant in an aqueous medium. To form secondary nanoparticles, and nanoparticles containing retinol or a derivative thereof obtained by allowing a monovalent to trivalent basic salt to act on the secondary nanoparticles;
(6) Nanoparticles containing retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant, at least partially or completely coated with a divalent or trivalent metal salt;
(7) The nanoparticles according to 4 above, further coated at least partially or completely with a monovalent to trivalent basic salt;
(8) The nanoparticle according to 4 above, wherein the aqueous medium is water;
(9) The nanoparticle according to the above 1, 2 or 4, wherein the divalent or trivalent metal salt is at least one selected from the group consisting of calcium salt, zinc salt, iron salt or copper salt;
(10) The monovalent to trivalent basic salt is one or more selected from the group consisting of hydrogen carbonate, hydrogen phosphate, carbonate, phosphate, oxalate, lactate, and urate. A nanoparticle according to 2 or 5 above;
(11) The nanoparticle according to the above 3, 4, 5 or 6, wherein the organic compound having a hydrophobic group and an anion residue is a fatty acid having 6 to 24 carbon atoms or a salt thereof;
(12) The nanoparticle according to 11 above, wherein the fatty acid having 6 to 24 carbon atoms or a salt thereof is one or more selected from the group consisting of an unsaturated fatty acid, a saturated fatty acid or a salt thereof;
(13) The nanoparticle according to 12 above, wherein the unsaturated fatty acid or a salt thereof is one or more selected from the group consisting of oleic acid, linoleic acid, linolenic acid or a salt thereof;
(14) The nanoparticle according to 12 above, wherein the saturated fatty acid or a salt thereof is at least one selected from the group consisting of lauric acid, myristic acid, palmitic acid, isostearic acid or a salt thereof;
(15) The surfactant is lecithin, polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan monolaurate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan Monopalmitate, polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (8) octylphenyl ether, polyoxyethylene (20) cholesterol ester, lipid-polyethylene glycol, polyoxyethylene hydrogenated castor oil and fatty acid-polyethylene glycol The nanoparticles according to the above-mentioned 3, 4, 5 or 6, which are one or more selected from the group consisting of polymers;
(16) The nanoparticle according to the above 1 to 15, wherein the particle has a diameter of 1 to 500 nm;
I will provide a.

Furthermore, the present invention provides, as another aspect,
(17) A skin or mucosa applicable external preparation containing the nanoparticles according to any one of 1 to 16 above;
(18) External preparations include ointments, gels, sublingual tablets, buccal tablets, liquids, sprays for oral and lower respiratory tracts, suction agents, suspensions, hydrogels, lotions, poultices, patches, The external preparation according to 17 above, which is selected from the group consisting of eye drops and nasal drops;
(19) An injectable preparation containing the nanoparticles according to any one of 1 to 16 above;
I will provide a.

Moreover, this invention is another aspect,
(20) Described below,
(A) a step of dissolving retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant in an organic solvent or a water-containing organic solvent;
(B) a step of dispersing the solution obtained in step (A) in water, and
(C) a step of mixing a divalent or trivalent metal salt with the solution obtained in step (B),
A method for producing nanoparticles comprising:
(21) The method for producing nanoparticles according to the above 20, further comprising (D) a step of mixing a monovalent to trivalent basic salt into the solution obtained in the step (C);
(22) The production method according to the above 20 or 21, wherein the organic solvent is at least one selected from the group consisting of acetone, ethanol, propanol, and butanol;
Is to provide.

The present invention is characterized in that retinol or a derivative thereof can be stably contained in a nanoparticle by mixing retinol or a derivative thereof with an organic compound having a hydrophobic group and an anion residue, which cannot be solubilized in an alkali. It has one characteristic.
In addition, the primary nanoparticles containing the retinol or derivative thereof thus obtained, and further the secondary nanoparticles and tertiary nanoparticles derived from the primary nanoparticles can have desired particle sizes. It has characteristics.

The nanoparticles provided by the present invention can absorb or retain retinol or a derivative thereof contained therein in a living body through the skin or mucous membrane, and can also be administered in vivo by injection administration. In any case, it exhibits excellent sustained release and targeting to the target action site.
Therefore, retinol or a derivative thereof, which has not been sufficiently achieved so far, has an epoch-making effect that enables percutaneous or transmucosal absorption in vivo, and has high absorption and sustained release. An external preparation and an injection containing the derivative are provided.

  Moreover, by transdermally administering the nanoparticles of the present invention from the epidermis to the deep part, retinol or a derivative thereof can be distributed at a high concentration in the dermis and subcutaneous tissue, so that it can be applied to cells forming the skin. It is useful for acting, and is extremely useful for improving skin troubles such as acne, pigmentation, fine wrinkles, and photoaging of the skin.

  Further, the nanoparticle of the present invention has the same effect on submucosa and can be applied to various diseases. Furthermore, it became possible to stabilize by adding retinol or a derivative thereof, which is less irritating but easily oxidized and low in stability, to the nanocapsule particles of the present invention. Therefore, the nanoparticles of the present invention can be used as pharmaceuticals, quasi drugs, and cosmetics.

  When nanoparticles containing retinol or a derivative thereof of the present invention are applied to the skin or mucous membrane, the penetration of the nanoparticles or the retinol or derivative contained therein into the skin is high, and the nanoparticles stay in a high concentration in the skin. Since it is easy, it can be expected that the effect of retinol or a derivative thereof on the skin is sustained.

In this specification, a simple notation “%” means “% (W / V)”.
Nanoparticles containing retinol or a derivative thereof provided by the present invention (hereinafter sometimes simply referred to as nanoparticles) have an average diameter (average particle diameter) of about 1 to 500 nm, preferably about 1 to It is 300 nm, more preferably about 10 to 200 nm. In the nanoparticles of the present invention, the average particle size can be selected according to the purpose, but from the viewpoint of stability and absorbability, 1 to 300 nm is preferable, and about 10 to 200 nm is more preferable.

  The particle size is determined by the blending ratio of the hydrophobic group and the organic compound having an anion residue to the retinol or its derivative to be contained, the addition amount of the surfactant, the addition amount of the basic salt, the amount of the solvent used, and the strength of stirring. It can be adjusted by. The particle diameter can be measured by a light scattering method or an electron microscope.

  Examples of retinol or a derivative thereof used in the present invention include various isomers of retinol and / or esters thereof. The isomer is preferably all-trans or 1,3-cis. More specifically, fatty acid esters such as retinol, retinol acetate, retinol butyrate, retinol propionate, retinol octylate, retinol laurate, retinol palmitate, retinol oleate, retinol linolenate, phosphate esters, phospholipid complexes, Examples include ethers with monohydric alcohols and ethers with polyhydric alcohols.

  Of these, retinol, retinol acetate, and fatty acid esters of retinol are preferable, and retinol, retinol acetate, and retinol palmitate are particularly preferable. Retinol or a derivative thereof may be not only one kind but also a mixture, and may be a fatty oil or an extract obtained from a marine animal tissue containing retinol or a derivative thereof.

  In the preparation of nanoparticles provided by the present invention, retinol or a derivative thereof can be added as it is, but it is preferable to use it dissolved in water, an organic solvent or a water-containing organic solvent. As such an organic solvent, lower alcohols such as acetone, methanol, ethanol, propanol and butanol can be used, and acetone and ethanol are particularly preferable. In the present invention, the content of retinol or a derivative thereof in the nanoparticles can be adjusted according to the purpose, but is usually 0.001% or more, preferably 0.005% or more, more preferably 0.01% or more. Especially preferably, it is 0.05% or more.

  In order to produce the primary nanoparticles of the present invention, an organic compound having an anion residue such as a carboxyl group, a phosphate group, and a sulfate group and a hydrophobic group such as an alkyl group is required, and these may be included. Any medium long chain organic compound having a carboxyl group as an anion residue is preferable. As the organic compound having an anion residue and a hydrophobic group, an unsaturated or saturated fatty acid having 6 to 24 carbon atoms or a salt thereof is more preferable. Specifically, unsaturated compounds such as oleic acid, linoleic acid, and linolenic acid are used. Fatty acids or salts thereof; saturated fatty acids such as lauric acid, myristic acid, palmitic acid, isostearic acid or salts thereof. Among these, oleic acid, myristic acid, isostearic acid or a salt thereof is particularly preferable. As the salt, potassium salt, sodium salt and the like can be used.

  These organic compounds having an anion residue and a hydrophobic group can be added as they are in the form of powder, but are preferably used by dissolving in water, an organic solvent or a water-containing organic solvent. As such an organic solvent, lower alcohols such as acetone, methanol, ethanol, propanol and butanol can be used, and acetone and ethanol are particularly preferable. As the water-containing organic solvent, those obtained by adding water to these organic solvents can be used, but the amount of water added is preferably about 80% by weight at the maximum.

  The weight ratio of retinol or a derivative thereof to the organic compound having an anion residue and a hydrophobic group is preferably about 1: 0.03 to 30, more preferably about 1: 0.05 to 20, and 1: 0. About 2 to 10 is particularly preferable.

  In order to obtain a good uniform state containing fine particles at the stage of producing these primary nanoparticles and secondary nanoparticles, by using a stirrer, probe type sonicator, etc. Fine primary nanoparticles and secondary nanoparticles can be produced by increasing the pressure using a cloak gorley or the like.

  In producing the primary nanoparticles of the present invention, it is preferable to add an appropriate amount of a surfactant in order to avoid aggregation of the produced nanoparticles. The blending amount can be appropriately selected as long as the nanoparticles do not aggregate with each other. The blending ratio of the organic compound having an anion residue and hydrophobic group to the surfactant is 1: 0.01 to a molar ratio. About 10 is preferable, and about 1: 0.05 to 5 is more preferable.

  As such surfactants, cationic surfactants, anionic surfactants, zwitterionic surfactants and nonionic surfactants can be used. Of these, nonionic surfactants are preferred from the viewpoint of low irritation to the skin or mucous membrane. Nonionic surfactants include polyoxyethylene (20) sorbitan monooleate (polysorbate 80), polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan monostearate (polysorbate). 60), sorbitan fatty acid esters such as polyoxyethylene (20) sorbitan monopalmitate (polysorbate 40), polyoxyethylene (20) sorbitan trioleate (polysorbate 85); polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil Polyoxyethylene castor oil / hardened castor oil derivatives such as polyoxyethylene glycerin monostearate, polyoxyethylene glycerin monoisostearate, etc. Steryls; glycerin fatty acids such as glyceryl monostearate; polyoxyethylene alkylphenyl ethers such as polyoxyethylene (8) octylphenyl ether; polyoxyethylene alkyl ethers such as polyoxyethylene 2-octyldodecyl ether; lecithin Lipid-polyethylene glycol; fatty acid-polyethylene glycol such as polyethylene glycol monolaurate, polyethylene glycol monooleate, polyethylene glycol monostearate; fatty acid-polyethylene glycol copolymer such as oleic acid-polyethylene glycol copolymer, etc. Can do.

  These surfactants can be used alone or in combination of two or more. In particular, lecithin, sorbitan fatty acid esters, polyoxyethylene castor oil / hardened castor oil derivative, and fatty acid-polyethylene glycol copolymer are preferable. More specifically, lecithin, polyoxyethylene (20) sorbitan monooleate, polyoxy Ethylene (20) sorbitan monolaurate and oleic acid-polyethylene glycol copolymer are preferred. Examples of fatty acids in this case include unsaturated fatty acids such as oleic acid, linoleic acid, and linolenic acid; and saturated fatty acids such as lauric acid, myristic acid, palmitic acid, and isostearic acid. As the amount of the surfactant is increased, the particle size is increased. However, if the amount is too small, aggregation between particles or large particles are formed.

  In preparing the primary nanoparticles of the present invention, vegetable oil may be further added. As the vegetable oil used in this case, vegetable oils such as soybean oil, sesame oil, corn oil, olive oil and various salad oils can be preferably used.

  The nanoparticles of the present invention include secondary nanoparticles obtained by reacting the primary nanoparticles obtained as described above with a divalent or trivalent metal salt, and these secondary nanoparticles are monovalent to trivalent basic. It consists of tertiary nanoparticles obtained by acting with salt.

  Examples of the divalent or trivalent metal salt for obtaining the secondary nanoparticles of the present invention include calcium salts such as calcium chloride, calcium acetate, and calcium sulfate; zinc salts such as zinc acetate, zinc chloride, and zinc sulfate; iron chloride Or an iron salt such as iron sulfide; or a copper salt such as copper chloride or copper sulfide, among which a calcium salt, particularly calcium chloride is preferred.

  This metal salt can also be used as a hydrate. The compounding amount of the metal salt cannot be generally limited, but the weight ratio of retinol or a derivative thereof and the metal salt is preferably about 1: 0.001 to 10, more preferably about 1: 0.002 to 5, A ratio of about 1: 0.01 to 5 is particularly preferable.

  In addition, monovalent to trivalent basic salts for obtaining tertiary nanoparticles include hydrogen carbonates such as sodium hydrogen carbonate and potassium hydrogen carbonate; hydrogen phosphates such as sodium hydrogen phosphate and potassium hydrogen phosphate. Salts; carbonates such as sodium carbonate, potassium carbonate and calcium carbonate; phosphates such as sodium phosphate, potassium phosphate and calcium phosphate; oxalates such as sodium oxalate, potassium oxalate and calcium oxalate; sodium lactate, Examples thereof include lactates such as potassium lactate and calcium lactate; urates such as sodium urate, potassium urate and calcium urate. Of these, hydrogen carbonate and carbonate are preferable, and sodium carbonate is particularly preferable. The blending amount of the basic salt cannot be generally limited, but the molar ratio of the divalent or trivalent metal salt to the basic salt is preferably about 1: 0.002 to 10, preferably 1: 0.001. About 5 is more preferable, and about 1: 0.02 to 2 is particularly preferable.

Below, the manufacturing method of the nanoparticle of this invention is demonstrated.
First, retinol or a derivative thereof, an organic compound having an anion residue and a hydrophobic group, and a surfactant are dissolved in an organic solvent or a water-containing organic solvent, and this solution is dispersed in a large amount of an aqueous medium, preferably water. Primary nanoparticles are produced by stirring this solution for about 1 to 30 minutes.

  A divalent or trivalent metal salt is allowed to act on the solution containing the primary nanoparticles thus prepared to prepare secondary nanoparticles. Specifically, a divalent or trivalent metal salt is added to a solution containing primary nanoparticles, and stirred for 1 to 30 minutes to prepare secondary nanoparticles.

  Next, a monovalent to trivalent basic salt is allowed to act on the solution of the secondary nanoparticles thus obtained to prepare tertiary nanoparticles. Specifically, tertiary nanoparticles can be obtained by adding a monovalent to trivalent basic salt to a solution of secondary nanoparticles and stirring for 1 minute to 24 hours.

  The aqueous medium used here is preferably water, but may be any water-containing organic solvent other than water, such as water-containing alcohol. In short, any aqueous medium that can be dispersed as nanoparticles may be used.

  The secondary or tertiary nanoparticle containing the retinol or derivative thereof of the present invention thus produced can be used as it is, or lyophilized, vacuum dried, spray dried, etc. Can be used as a dry product after removing the solvent. By using these solutions or dried products, formulation bases, additives and the like as appropriate, it is possible to prepare a desired preparation for parenteral administration, an external preparation for skin or mucosa or an injection for injection.

  The present invention also provides such an external preparation or injectable preparation for skin / mucosa application. Examples of such external preparations are those that can be administered in the form of local application, application, application, dripping, spraying, etc. for the purpose of systemic and local administration / treatment. Specifically, ointments, gels, sublingual agents Oral tablets, liquids, sprays for oral cavity and lower respiratory tract, suction agents, suspensions, hydrogels, lotions, poultices, patches and the like. As the solution, nasal drops and eye drops are preferable.

  In addition, application to the skin or mucous membrane, spraying to the lower respiratory tract, etc. are effective dosage forms. As an injectable agent, any of administration forms such as intravenous injection, subcutaneous injection and intramuscular injection is possible.

  Examples of these external preparations, bases used for the preparation of injections, and other additive components include bases and components that are pharmaceutically used for preparing external preparations and injections. Specifically, oily bases such as petrolatum, plastibase, paraffin, liquid paraffin, light liquid paraffin, honey beeswax, silicone oil; water, water for injection, ethanol, methyl ethyl ketone, cottonseed oil, orb oil, peanut oil, sesame oil, etc. Solvent: Polyvinylpyrrolidone, sodium carboxymethylcellulose, xanthan gum, tragacanth gum, gum arabic, gelatin, albumin and other thickeners; stabilizer such as dibutylhydroxytoluene; glycerin, 1,3-butylene glycol, propylene glycol, urea, sucrose Moisturizers such as erythritol and sorbitol; preservatives such as methyl paraoxybenzoate, butyl paraoxybenzoate, sodium dehydroacetate, p-cresol, etc. Door can be.

  In the case of nasal drops, it is preferable to add a nasal absorption promoter such as hydroxypropylcellulose. Furthermore, it is good to use gelling agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, polyvinylpyrrolidone, to make a hydrogel agent. For example, in the case of an ointment containing the nanoparticles of the present invention as an active ingredient, 0.05 to 0.5% sodium carboxymethylcellulose is used to stabilize the suspension using petrolatum as a component such as a base. Should be used together.

  Examples of the present invention and test examples are described below, but the present invention is not limited to these examples.

Example 1 Preparation of Nanoparticles 50 μL of an aqueous sodium oleate solution (100 mg / mL) was added to 850 μL of purified water and stirred. Further, 20 μL of an aqueous solution of polysorbate 80 (50 mg / mL) and 50 μL of an ethanol solution of retinol (20 mg / mL) were added and stirred to prepare a solution containing primary nanoparticles. To this solution containing primary nanoparticles, 25 μL of 1M calcium chloride aqueous solution was added and stirred to prepare a solution containing secondary nanoparticles. Further, 5 μL of 1M sodium hydrogen carbonate aqueous solution was added to prepare a solution containing tertiary nanoparticles.

When the particle size of the obtained particles was measured with a dynamic light scattering particle size distribution analyzer (NICOMP 380ZLS, US Particle Sizing Systems), the average particle size of the tertiary nanoparticles was 9.22 nm. Further, the mixture was centrifuged at 10,000 rpm for 10 minutes, and the retinol concentration in the supernatant (tertiary nanoparticle solution) was quantified according to a conventional method using high performance liquid chromatography. It was 80.8%. The retinol content of the tertiary nanoparticles was calculated by the following formula.
Retinol content (%) = (retinol concentration in supernatant × supernatant volume) / input amount × 100

Test Example 1 : Effects of calcium and surfactant on nanoparticle formation A 5.0 mg / mL sodium oleate aqueous solution, a calcium chloride aqueous solution and a polysorbate 80 aqueous solution described in Tables 1 and 2 below were prepared. According to the method, tertiary nanoparticles were prepared, and the particle diameter and the retinol content of the tertiary nanoparticles were measured.
The results are shown in Tables 1 and 2 below.

* 1: The molar ratio of oleic acid to polysorbate 80 ([Polysorbate 80] / [Oleic acid]) is 0.93, 0.47, 0.00 when the polysorbate 80 aqueous solution concentration is 2, 10, and 20 mg / mL, respectively. 93.

  As can be seen from the results shown in the table, it was confirmed from the measured values of retinol content and particle size that tertiary nanoparticles containing retinol were produced under any conditions.

Examples 2 to 4 / Comparative Examples 1 and 2 : Preparation of tertiary nanoparticles Tertiary nanoparticles were prepared according to the formulation shown in Table 3 below. The preparation of the tertiary nanoparticles was carried out according to the preparation method of Example 1, and after dissolving sodium oleate or sodium laurate in purified water, polysorbate 80 or polysorbate 20 and retinol dissolved in ethanol were stirred, and 1M Prepared by adding calcium chloride and 1M sodium bicarbonate sequentially with stirring.

Test Example 2 : Stability Test of Retinol-Containing Nanoparticles In the tertiary nanoparticle solutions of Examples and Comparative Examples prepared according to the formulation in Table 3, the average particle size, turbidity, and retinol content were measured immediately after preparation and at 40 ° C. for 14 days. Measured after storage. The average particle size is measured with a dynamic light scattering particle size distribution analyzer, the turbidity is measured at an absorbance of 550 nm, and the retinol content is determined by quantifying the retinol content according to a conventional method using high performance liquid chromatography. did.
The results are summarized in Table 4 below.

  As can be seen from the results shown in the table, the storage stability of retinol was remarkably improved by using the nanoparticles of the present invention.

Test Example 3 : Absorbency test of nanoparticles The absorbability of the tertiary nanoparticles of the present invention into the skin was evaluated.
First, a 6-week-old hairless mouse (female, HR-1 strain, Japan SLC Co., Ltd.) was euthanized, and then the skin was removed from the back. This was attached to a plate membrane permeation experiment apparatus (capacity: 5 mL, effective area: 0.64 cm ² , manufactured by Vidrex), and the receptor (dermis side) solution was filled with 50% ethanol. The inside of the cell was kept constant at 37 ° C. by refluxing water to the water jacket of the flat membrane permeation test apparatus, and the samples of Example 1 and Comparative Example 1 were used as specimens, and 5 mL each was placed on the donor (keratin layer) side. An absorption test was conducted. After 24 hours, the skin was removed from the permeation test apparatus for flat membranes, washed with purified water to remove retinol adhering to the surface of the skin, homogenized skin, and extracted with retinol to contain retinol in the skin. The amount was quantified by liquid chromatography according to a conventional method.
The results are shown in Table 5 below.

  As can be seen from the results shown in the table, it is understood that the absorption of retinol into the skin is remarkably improved by using the nanoparticles of the present invention.

Example 5 / Comparative Example 3 External Cream According to Table 6 below, the oil phase components (1) to (5) were mixed and dissolved, and kept at 75 ° C. Separately, the aqueous phase components (6) to (7) were mixed and kept at 75 ° C. The mixture of the oil phase components is added to the mixture of the water phase components, uniformly emulsified with a homomixer, cooled to 40 ° C., and then the components (8) to (9) are added thereto. The mixture was uniformly mixed to obtain a skin cream.

  As a result of using the external skin cream obtained in Example 5 above for 3 months on a subject who showed significant wrinkle formation on the face, an effect of improving wrinkles was observed.

  From the above results, retinol was stably contained in the nanoparticles of the present invention, and high transdermal absorbability was confirmed. In the external preparation for skin containing the retinol nanoparticles of the present invention, more retinol can be delivered into the skin, so that it was possible to sufficiently exhibit effective actions such as the anti-wrinkle effect of retinol.

As described above, the present invention provides nanoparticles containing retinol or a derivative thereof having excellent transdermal absorbability through skin or mucosal administration for systemic and local administration.
The nanoparticle of the present invention penetrates well from the epidermis to the deep part by transdermal administration, and can distribute retinol or its derivative in a high concentration in the dermis and subcutaneous tissue, thus acting on the cells forming the skin. Useful for making
Therefore, the external preparation containing the nanoparticles of the present invention is extremely useful for improving skin troubles such as acne, pigmentation, fine wrinkles, and photoaging of the skin, and its industrial applicability is great. Is.

Claims (22)

  Nanoparticles obtained by reacting primary nanoparticles containing retinol or a derivative thereof with a divalent or trivalent metal salt.   Making primary nanoparticles containing retinol or its derivatives into secondary nanoparticles by acting with a divalent or trivalent metal salt, and allowing the monovalent to trivalent basic salt to act on the secondary nanoparticles. Nanoparticles consisting of   The nanoparticle according to claim 1 or 2, wherein the primary nanoparticle comprises a retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant.   A divalent or trivalent metal salt is allowed to act on primary nanoparticles prepared by dispersing retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant in an aqueous medium. Nanoparticles containing retinol or a derivative thereof.   A divalent or trivalent metal salt is allowed to act on primary nanoparticles prepared by dispersing retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant in an aqueous medium. Secondary nanoparticles, and nanoparticles containing retinol or a derivative thereof obtained by allowing a monovalent to trivalent basic salt to act on the secondary nanoparticles.   A nanoparticle comprising a retinol or derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant, at least partially or completely coated with a divalent or trivalent metal salt.   The nanoparticle according to claim 4, which is further at least partially or completely coated with a monovalent to trivalent basic salt.   The nanoparticle according to claim 4, wherein the aqueous medium is water.   The nanoparticle according to claim 1, 2, or 4, wherein the divalent or trivalent metal salt is at least one selected from the group consisting of calcium salts, zinc salts, iron salts and copper salts.   The monovalent to trivalent basic salt is at least one selected from the group consisting of hydrogen carbonate, hydrogen phosphate, carbonate, phosphate, oxalate, lactate and urate. The nanoparticle according to 2 or 5.   The nanoparticle according to claim 3, 4, 5, or 6, wherein the organic compound having a hydrophobic group and an anion residue is a fatty acid having 6 to 24 carbon atoms or a salt thereof.   The nanoparticle according to claim 11, wherein the fatty acid having 6 to 24 carbon atoms or a salt thereof is one or more selected from the group consisting of an unsaturated fatty acid, a saturated fatty acid, or a salt thereof.   The nanoparticle according to claim 12, wherein the unsaturated fatty acid or a salt thereof is at least one selected from the group consisting of oleic acid, linoleic acid, linolenic acid or a salt thereof.   The nanoparticle according to claim 12, wherein the saturated fatty acid or a salt thereof is at least one selected from the group consisting of lauric acid, myristic acid, palmitic acid, isostearic acid or a salt thereof.   Surfactant is lecithin, polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan monolaurate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan monopalmitate , Polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (8) octylphenyl ether, polyoxyethylene (20) cholesterol ester, lipid-polyethylene glycol, polyoxyethylene hydrogenated castor oil and fatty acid-polyethylene glycol copolymer The nanoparticle according to claim 3, 4, 5, or 6, which is one or more selected from the group consisting of:   The nanoparticle according to any one of claims 1 to 15, wherein the particle has a diameter of 1 to 500 nm.   An external preparation for skin or mucosa application comprising the nanoparticles according to any one of claims 1 to 16.   External preparations include ointments, gels, sublingual tablets, buccal tablets, solutions, sprays for oral and lower respiratory tracts, suction agents, suspensions, hydrogels, lotions, poultices, patches, eye drops and The external preparation according to claim 17, which is selected from the group consisting of nasal drops.   An injectable preparation comprising the nanoparticles according to any one of claims 1 to 16. (A) a step of dissolving retinol or a derivative thereof, an organic compound having a hydrophobic group and an anion residue, and a surfactant in an organic solvent or a water-containing organic solvent;
(B) a step of dispersing the solution obtained in step (A) in water, and
(C) a step of mixing a divalent or trivalent metal salt with the solution obtained in step (B),
A process for producing nanoparticles comprising   21. The method for producing nanoparticles according to claim 20, further comprising (D) a step of mixing a monovalent to trivalent basic salt into the solution obtained in step (C). The production method according to claim 20 or 21, wherein the organic solvent is one or more selected from the group consisting of acetone, ethanol, propanol, and butanol.