KR101172066B1 - Lipid-based drug delivery carrier containing stigmasteryl hemisuccinate and the composition of skin external application containing the same - Google Patents

Abstract

The present invention relates to a lipid-based drug carrier comprising stigmasteryl hemisuccinate and a topical skin composition containing the same, more particularly liposomes prepared using stigmasteryl hemisuccinate or salts thereof, micro Lipid-based drug carrier and external skin composition containing the same as an emulsion or nanoemulsion, which has a sensitive sensitivity to changes in acidity to effectively release in vivo the bioactive components incorporated into the inner phase to enhance the effect as a carrier. It is about.

Stigmasteryl Hemisuccinate * Acidity Sensitivity * Liposome * Microemulsion * Nanoemulsion * Drug Carrier

Description

Lipid-based drug delivery carrier containing stigmasteryl hemisuccinate and the composition of skin external application containing the same}

1 shows the size distribution of liposomes prepared in Examples 4 and 7 as a result of Test Example 1 in (a) and (b), respectively.

Figure 2 shows the change in turbidity according to the change in acidity of the liposome dispersion prepared in Example 4 as a result of Test Example 2.

FIG. 3 is a result of Test Example 3, prepared to support HPTS in each of the liposomes prepared in Example 4 and the control group of existing acidity-sensitive liposomes, and the amount of internally supported initial HPTS was measured at pH 7. The initial content is shown in (a) and the initial content measured at pH 3 is shown in (b).

4 is a result of Test Example 3, so as to support the HPTS on the liposome prepared in Example 4 added to the MES buffer (pH 3) and HEPES buffer solution (pH 7), and then sampled at a predetermined time liposomes The amount of HPTS emitted to the outside is shown by measuring with a fluorescence spectrophotometer (Fluorescence Spectrophotometer).

5 is a result of Test Example 4, after incorporating BSA into the inner liposomes prepared in Examples 4 and 9 and the respective control liposomes, and measured and compared the amount of BSA initially included (a: Example 4, b: control, c: Example 9, d: control).

FIG. 6 is a result of Test Example 5, incorporating a whitening effector 0.5mM and 0.25mM actinomycetes culture extract into the liposomes of Example 4, and adding it to the cell culture medium to use it in cell culture and the amount of melanin production (b ) And melanin production in cells cultured with culture medium containing only 1mM, 0.5mM and 0.25mM concentrations of Actinomycetes culture extracts are shown in (a), respectively.

7 is a result of Test Example 5, incorporating a whitening effector 0.5mM actinomycetes culture extract on the inside of the liposome of Example 4, and added to the cell culture medium (a), the control was loaded with 0.5mM actinomycetes culture extract Existing acidity-sensitive lipid constructs were added to the cell culture medium (b) and cultured for 5 days each, followed by optical microscopy of the changed properties of the cells.

8 is prepared as a result of Test Example 6, the casein is encapsulated in the liposomes of Example 4 and treated for 4 hours and then quantified the amount of calcein introduced into the cell by flow cytometry in (b) In addition, liposomes consisting of phosphatidylcholine: cholesterol carrying a control calcein were treated for the same time, and then the amount of intracellular import of calcein was quantified and shown in (a).

The present invention relates to a lipid-based drug carrier comprising stigmasteryl hemisuccinate and a topical skin composition containing the same, more particularly liposomes prepared using stigmasteryl hemisuccinate or salts thereof, micro Lipid-based drug carrier and external skin composition containing the same as an emulsion or nanoemulsion, which has a sensitive sensitivity to changes in acidity to effectively release in vivo the bioactive components incorporated into the inner phase to enhance the effect as a carrier. It is about.

Recently, as the usefulness of drug delivery systems using liposomes has emerged, researches for producing more effective lipid-based nanocarriers through mixing various lipid components have been attracting attention. In particular, in order to ensure that the physiologically active ingredient incorporated into the liposome inner phase can be delivered to a living cell with high efficiency to maximize the efficacy of the active ingredient, studies are being actively conducted to prepare various components of the liposome. For example, to prepare liposomes containing lipids into which biocompatible polymers are introduced to increase the in vivo circulation time to enhance the bioavailability of the active drug; Liposomes including lipids into which cell recognition or cell adhesion molecules have been introduced to specifically deliver large molecular weight drugs such as peptides, proteins or genes to target cells; Or research on liposomes designed to release the physiologically active ingredients embedded in liposomes by reducing the stability of liposomes by the specific environment of the intracellular organelles.

In particular, various systems have been attempted to release the bioactive components incorporated into the internal organs in response to changes in the acidity environment of intracellular organelles. For example, dioleoylphosphatidylethanolamine (“DOPE”) vesicles stabilized by the introduction of a pH-sensitive co-surfactant has been studied extensively in acidic solutions at pH 4.5-6.5 (FC). Szoka et al ., J. Liposome Res . (1994) 4, 361), in fact the delivery of vesicles to the cytoplasm of various cells has been confirmed.

However, formulations made at the beginning of the liposome study show low stability in blood and have many limitations in direct application in vivo. In most cases, as a result of basic research for gene delivery, the production of delivery material and delivery efficiency for practical use could not be solved, and thus, the practical use in the industry was not achieved.

Thus, the present inventors have made intensive studies for the purpose of improving the delivery and release of bioactive substances in vivo, and for the purpose of commercialization and formulation of existing liposomes, in the preparation of lipid-based nanocarriers such as liposomes and nanoemulsions, When stigmasteryl hemisuccinate is used in combination, the resulting lipid membrane is stabilized to enhance the amount of drug that can be supported in the system, and is sensitive to specific changes in the acidity environment of tissue organelles in the body. It was found that the present invention was completed.

In other words, the present inventors have found that liposomes or nanoemulsions of the present invention prepared using stigmasteryl hemisuccinate (or "Stigmaster-5,22-diene-3-ol, Hydrogen Butanedioate") Acidity-sensitive lipid-based nanocarriers capable of controlling the release of physiologically active ingredients incorporated into the inner phase and delivery enhancing formulations, which are not easily delivered in vivo due to relatively large molecular weight, chemical structure, or insolubility. It moves to the target organs stably incorporating into the target organs, and in response to specific changes in the acidity environment of the target organs, it is possible to enhance the effect as a carrier by disintegrating the lipid membrane and releasing physiologically active ingredients. It has been found that can be provided to the maximum.

In the present invention, to provide a lipid-based nanocarrier prepared to enhance the delivery efficiency in vivo by enabling the release control of the bioactive components incorporated into the inner phase.

In another aspect, the present invention is to provide a lipid-based nanocarrier having a property to stabilize the phospholipid membrane constructs and to respond to changes in acidity environment.

Accordingly, it is an object of the present invention to provide an acidity-sensitive lipid-based nanocarrier prepared using stigmasteryl hemisuccinate and incorporating a bioactive component into the inner phase.

The present invention also provides an external composition for skin containing the acidity-sensitive lipid-based nanocarrier as an active ingredient.

In the present invention, as a lipid-based nanocarrier having a form of liposomes or nanoemulsions, in addition to the lipid component in the lipid-based structure, Stigmasteryl hemisuccinate (hereinafter referred to as "SHEMS") ), And the inner phase provides lipid-based nanocarriers with bioactive components:

The stigmasteryl hemisuccinate is derived from a carboxyl group at the end of the stigmasterol, the SHEMS alone; Or one selected from salts of SHEMS such as tris salt, tris (hydroxymethyl) aminomethane salt and cyclohexyl ammonium salt thereof; Alternatively, a mixture of SHEMS and two or more selected from the salts of SHEMS may be used in combination with a lipid, which may be appropriately selected depending on the characteristics of the carrier and the manufacturing process.

Lipid-based nanocarriers according to the invention can be prepared in the form of liposomes or nanoemulsions. In preparing such liposomes or nanoemulsions, the lipid membrane structure can be stabilized by using the above-described SHEMS as a component of the lipid membrane structure. On the other hand, the lipid-based nanocarrier is sensitive to acidity, so that the stability of the lipid membrane in the region below the acidity 6.0 is lowered, and at a lower acidity is disintegrated to release the physiologically active component embedded in the inner phase.

As a result, the lipid-based nanocarrier can effectively release the biologically active ingredient contained in the inner box in accordance with the change in acidity in vivo, thereby enabling the preparation of an external preparation for skin that can maximize the efficacy of the biologically active ingredient. It may also be prepared in oral dosage form.

Therefore, it is also within the scope of the present invention to provide an external composition for skin or a composition for oral administration having such properties.

In the present invention, the lipid-based nanocarrier is prepared in the form of liposomes or nanoemulsions, and as the lipid component of the lipid construct, phospholipids or nitrogenous lipids having 12 to 24 fatty acid chains may be used. It is generally more preferable to use phospholipids, for example, egg yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidine acid, phosphatidylserine, phosphatidylglycerol, phosphatidylglycerol Natural phospholipids such as inositol, phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, and plasmalogen; Hydrogenated products obtainable from these phospholipids by conventional methods; And dicetyloylphosphate, distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostaroylphosphatidylcholine, eleostaroylamine Synthetic lipids such as eleostaroylphosphatidylserine; Derivatives thereof; And fatty acid mixtures obtainable by their hydrolysis.

Lipids containing phospholipids may be used alone or in combination of two or more. When two kinds of phospholipids are mixed and used, Preferably, for example, the following combinations; Phosphatidylcholine: phosphatidylethanolamine, phosphatidylcholine: phosphatidylglycerol, phosphatidylcholine: phosphatidyl inositol, phosphatidylcholine: phosphatidine acid or phosphatidylcholine: dioleoyl phosphatidylethanolamine and the like. These mixing ratios differ depending on the component composition to be mixed, and the mixing ratio of the maximum component to the minimum component is preferably 1: 5 or less. For example, in the case of phosphatidylcholine: dioleoylphosphatidylethanolamine, the molar ratio of 5: 1 to 1: 5 is 1: 1, 2: 1, 3: 1, 4: 1, 5: 1, 1, respectively. It can be produced in various ways such as: 5, 1: 4, 1: 3 or 1: 2. In addition, even when three kinds of phospholipids are used in combination, for example, phosphatidylcholine: dioleoylphosphatidylethanolamine: phosphatidylserine in a molar ratio range of 1: 5, respectively, 1: 1: 1, 2: 1: 1, 3: 1: 2, 3: 2: 1. 3: 2: 2. It can be produced in various mixing ratios such as 4: 1: 1 or 4: 2: 1.

In addition, the molar ratio of the lipids: SHEMS is preferably 9: 1 to 1: 9, more specifically 9: 1, 4: 1, 3: 1, 7: 3, 2: 1, 3: 2, It can be produced in various ways, such as 2: 3 or 1: 4, but is not limited thereto.

The lipid component of the nanocarrier is used in an amount of 0.001 to 20% by weight, preferably 0.2 to 10% by weight, based on the total amount of liposome dispersion or nanoemulsion. Hereinafter, the lipids used with SHEMS in the present invention will be mainly described by phospholipids.

In the present invention, additives such as excipients or stabilizers other than stigmasteryl hemisuccinate may be used in consideration of the stability and operability of the liposome preparation.

The biologically active ingredient incorporated into the nanocarrier inner phase of the present invention is not limited to its kind such as water-soluble or water-insoluble as long as it can be applied to a living body. For example, it may be a single component or an animal or plant or strain extract, and a mixture containing two or more thereof, which may be used to adjust according to the purpose and case. As an ingredient that enhances whitening or has an effect of preventing wrinkles and aging and treating, preferably, N-butyldeoxynojirimycin, 1-dioxynojirimycin, castanospermine, actinomycete culture Extract (SCE) or Retinol may be used. These bioactive components may be incorporated in amounts of 0.01-30% by weight, preferably 0.1-20% by weight, based on the total weight of the liposome dispersion or nanoemulsion.

As a method of forming a nanocarrier containing a physiologically active component using the lipid component added SHEMS proposed in the present invention, the phospholipids and SHEMS are dissolved in an organic solvent, and then the solvent is evaporated and decompressed to form a lipid film. Adding an aqueous solution and then applying ultrasonic waves; Dispersing phospholipids and SHEMS dissolved in an organic solvent in an aqueous solution and then applying ultrasonic waves; Dispersing or dissolving phospholipids and SHEMS in an organic solvent and then extracting or evaporating the organic solvent with excess water; Dispersing or dissolving phospholipids and SHEMS in an organic solvent, then vigorously stirring using a homogenizer or a high pressure emulsifier and evaporating the solvent; Dispersing or dissolving phospholipids and SHEMS in an organic solvent and dialysis with excess water; And a method of gradually adding water after dispersing or dissolving phospholipids and SHEMS in an organic solvent.

In relation to the above production method, in order to dissolve the phospholipids and SHEMS in an organic solvent, a mechanical force may be applied or heated to 20 to 100 ° C, preferably 70 ° C or less. When using the salt of SHEMS, the salt of SHEMS can be dissolved in an aqueous solution, and can be used.

When the bioactive ingredient is water-soluble, the bioactive ingredient is dissolved in water or dissolved in an aqueous solution, and then administered together in the step of adding the aqueous solution or water. When the physiologically active component is water insoluble, the physiologically active component may be prepared by dissolving the physiologically active component in an organic solvent and then including the lipid component in an organic solvent phase.

Organic solvents that can be used to dissolve phospholipids and SHEMS or water-insoluble physiologically active ingredients in the above method include acetone, dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, dioxane, tetrahydrofuran, acetic acid , A solvent selected from ethyl acetate, acetonitrile, methyl ethyl ketone, methylene chloride, chloroform, methanol, ethanol, ethyl ether, diethyl ether, hexane and petroleum ether, or a mixture thereof can be used.

The nanocarrier of the present invention obtained by the above-described manufacturing method has an average particle diameter of 10 to 1,000 nm, more preferably 10 to 500 nm, depending on the composition and the preparation method of the lipid membrane, and the acidity as confirmed in the test example described later. It has a sensitive sensitivity to change, thereby effectively releasing the physiologically active component incorporated into the internal organs in the cell to enhance its effect as a transporter.

Therefore, it can function as a transporter that can double the physiological activity of the physiologically active component, for example, the physiologically active component having the effect of whitening enhancement or wrinkle and anti-aging and treatment, and the particle size of the colloidal stability It has excellent chemical stability and can be used in various external skin preparations. In addition, as a phospholipid construct is harmless to the human body can be usefully used to increase the bioavailability of the pharmaceutical composition or health food composition for oral administration.

The external preparation composition for skin containing the nanocarrier of the present invention is not particularly limited in its formulation, and it is a softening longevity, astringent longevity, nutrient longevity, nutrition cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing Lotion, cleansing foam, cleansing water, pack, powder, makeup base, foundation, body lotion, body cream, body oil, body essence, body cleanser, hair dye, shampoo, rinse, toothpaste, mouthwash, hair conditioner, wool, lotion , Ointments, gels, creams, patches or sprays and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples, and the materials, reagents, costs, and operations described in these Examples and Test Examples may be appropriately changed or selected within the scope of the present invention.

In addition, these Examples are only illustrative description of this invention, and the scope of the present invention should not be interpreted as being limited to these Examples.

<Examples 1-9>

Phosphatidylethanolamine (phospholipid) and SHEMS were mixed in a molar ratio of 3: 2 and dissolved in a chloroform: methanol (95: 5, volume / volume) mixed organic solvent, and the solvent was evaporated under reduced pressure to form a film. To the film from which the organic solvent was removed, 0.5 mM and 0.25 mM actinomycetes culture extracts dissolved in purified water as physiologically active ingredients were added, followed by ultrasound to prepare a liposome dispersion. Specifically prepared according to the ratio of Table 1.

In Examples 1 to 9, first, the nanocarriers were prepared using actinomycetes culture extracts as physiologically active ingredients. In addition, for the preparation and comparison of various cosmetics, in the following test examples, other than the above actinomycetes culture extract, other whitening agents, such as castanospermine, N-butyldioxy nozirimycin or 1-deoxy nojirimycin; And liposome dispersions according to the preparation methods and preparation conditions of Examples 1, 4, 7, and 8 below using calcein (calcein) or HPTS (8-hydroxypyrene-1,3,6-trisulfonic acid) as a fluorescent labeling substance. Prepared. In preparing the following examples, the lipid component content in the lipid dispersion was prepared at 0.2, 2, 5 and 10% by weight, respectively. The lipid-based carriers used in the following test examples were all prepared and used with a lipid component content of 0.2% by weight.

Liposome Preparation Conditions According to the Ratio of Phospholipids and SHEMS Phospholipids Phospholipids: SHEMS molar ratio Lipid content in lipid dispersion (% by weight) Example 1 Phosphatidylethanolamine 3: 2 0.2, 2, 5, 10 Example 2 Dioleoylphosphatidylethanolamine 8: 2 0.2, 2, 5, 10 Example 3 Dioleoylphosphatidylethanolamine 7: 3 0.2, 2, 5, 10 Example 4 Dioleoylphosphatidylethanolamine 6: 4 0.2, 2, 5, 10 Example 5 Dioleoylphosphatidylethanolamine 4: 6 0.2, 2, 5, 10 Example 6 Dioleoylphosphatidylethanolamine 2: 8 0.2, 2, 5, 10 Example 7 Phosphatidylcholine 3: 1 0.2, 2, 5, 10 Example 8 Phosphatidylcholine: dioleoylphosphatidylethanolamine (1: 1) (1: 1)
(1: 1) 1 ((1: 1) 2: 1 0.2, 2, 5, 10 Example 9 Phosphatidylcholine: dioleoylphosphatidylethanolamine (2: 3) 3: 2 0.2, 2, 5, 10

<Examples 10-16>

Saturated lecithin (Lipoid S75-3) extracted from soybean, rapeseed sterol (PEG-5 rapeseed Sterol; Generol RE 5) added with 5 mol of polyethylene glycol, ethyl hexanediol and alcohol or propylene glycol are shown in Table 2 below. Purified water containing 0.4% by weight of actinomycetes culture extract based on disodium EDTA, PEG 75, triethanolamine, and bioactive components, which were heated to 60 ° C, and dissolved at a temperature of 60 ° C. It was mixed with and emulsified at 5,000 rpm for 10 minutes with a homogenizer. Then, using a high-pressure emulsifier (Microfluidizer) three times room temperature recycling at 1,000 atmospheres to prepare a nanoemulsion.

In Examples 10 to 16, the nanocarriers were prepared using the actinomycetes culture extract as a physiologically active ingredient for the preparation of the following formulation examples and the efficacy test of the lipid-based nanocarriers. On the other hand, for the preparation and comparison of various cosmetics, in the following test examples, other whitening agents other than the above actinomycetes culture extract, such as castanospermine, N-butyl dioxy nozirimycin and 1-dioxy nojirimycin is used The nanoemulsion was prepared according to the preparation methods and preparation conditions of Examples 12 and 16.

 Nanoemulsion Manufacturing Conditions ingredient Example (% by weight) 10 11 12 13 14 15 16 Lecithin (Lipoid S75-3) 2.0 2.0 2.0 2.0 2.5 2.5 2.5 SHEMS 1.5 1.0 0.5 0.25 1.0 0.7 0.5 PEG-5 Rape Seed Sterol 0.5 1.0 0.5 1.5 0 0.3 0.5 Ethyl hexanediol (EHDIOL) 1.0 1.0 1.0 1.0 1.0 1.0 1.0 ethanol 10.0 10.0 10.0 10.0 0 0 0 Propylene Glycol (P.G.) 0 0 0 0 15.0 15.0 15.0 Purified water Balance Balance Balance Balance Balance Balance Balance EDTA? 2Na 0.05 0.05 0.05 0.05 0.05 0.05 0.05 PEG75 (PEG4000) 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Triethanolamine (TEA) 0.05 0.05 0.05 0.05 0.07 0.07 0.07

Test Example 1 Measurement of Size by Dynamic Light Scattering of Lipid-Based Nanocarriers

The average particle size of liposomes and nanoemulsions prepared in Examples 1-16 was measured using dynamic laser scattering (Dynamic light scattering, instrument model Zetasizer 3000HS, Malvern, UK). The scattering angle was fixed at 90 ° and the temperature was maintained at 25 ° C., and the average particle diameter was calculated from the Stokes-Einstein equation, and the results are shown in Table 3. The liposomes of Examples 1 to 9 showed only the average particle size when the liposome was prepared with a lipid component ratio of 0.2% by weight.

Nanocarrier average particle size prepared in Examples 1-16 Liposome Average diameter (nm) Nano Emulsion Average diameter (nm) Example 1 184.2 Example 10 101.2 Example 2 195.3 Example 11 100.8 Example 3 200.6 Example 12 102.8 Example 4 205.4 Example 13 98.4 Example 5 182.3 Example 14 105.2 Example 6 178.8 Example 15 99.6 Example 7 182.1 Example 16 104.5 Example 8 178.5 Example 9 175.6

On the other hand, the size distribution of the liposomes prepared in Examples 4 and 7 to confirm the particle distribution of liposomes as a nanocarrier according to the present invention is shown in FIG.

<Test Example 2> Sensitivity of SHEMS-Introduced Lipid-based Nanocarriers to Acidity Environmental Changes

In order to test the sensitivity of the lipid-based nanocarriers introduced SHEMS provided by the present invention according to the change in the acidity environment, the change in the acidity of the liposome dispersion prepared in Example 4 was observed to visually observe the change in the properties of each liposome The results are shown in FIG.

The nanocarrier dispersion prepared in Example 4 had an increase in turbidity from acidity 6.0 and slowly began to disintegrate with lipid structure no longer stable from acidity 4.5, and at acidity 2.5 it was observed that the lipid structure completely collapsed.

Through this test, the lipid-based nanocarrier introduced with the SHEMS provided by the present invention reacts specifically in the general acidity range 6.0-4.5 of the early endosomes, which are organelles of cells, thereby rapidly decreasing the stability of the lipid construct. It was confirmed that the delivery effect as a transporter could be enhanced by effectively releasing the physiologically active ingredient contained in the cell to reduce the activity of the physiologically active ingredient.

Test Example 3 Effect of Controlled Release by SHEMS-Incorporated Lipid-Based Nanocarriers

In order to confirm the release control effect by the acidity change of the lipid-based nanocarrier introduced SHEMS provided in the present invention, prepared in Example 4 using a 10 mM HEPES solution (pH 7.4) dissolved in 17.5 mM HPTS Liposomes were used. In order to confirm the initial amount of HPTS embedded in the liposomes in Example 4, the prepared liposome dispersion was used to remove HPTS not present in the liposomes using a Sephadex ^™ G-25 column. The liposome dispersion from which free HPTS has been removed is dispersed in MES buffer (pH 3) and HEPES buffer (pH 7), and the dispersion is again treated with 10% Triton X-100 to completely dissolve lipid-based structures in the dispersion. The decay was carried out so that all of the HPTS contained therein was released to the outside, and then quantified by Fluorescence Spectrophotometer (Hitachi F-4500).

In the same manner as above, liposomes composed of dioleoylphosphatidylethanolamine were prepared using a 10 mM HEPES solution (pH 7.4) in which 17.5 mM HPTS was dissolved in a film-hydration method. Was prepared. The prepared liposome dispersion was measured as the initial HPTS amount internally supported in the same manner as the control, the initial content at pH 7 is shown in Figure 3a, the initial content at pH 3 is shown in Figure 3b, respectively.

In addition, in order to confirm the effect of controlling the amount of release over time, the lipid-based liposome dispersion of SHEMS introduced in Example 4, in which the free HPTS present in the outer phase was removed using a Sephadex ^TM G-25 column, was used in MES buffer. The amount of HPTS released to lipid-based nanocarriers into which SHEMS was introduced by adding to (pH 3) and HEPES buffer (pH 7) and sampled every hour was measured with a fluorescence spectrophotometer (Hitachi F-4500). The amount of HPTS released with time and acidity change is shown in FIG. 4.

Through this test, it was confirmed that the lipid-based nanocarrier in which the SHEMS provided by the present invention is introduced can enhance the amount of drug that can be supported in the lipid-structure compared to the existing acidity-sensitive lipid-structure. In addition, since the amount of the drug released at pH 3 is significantly improved than at pH 7, it was confirmed that the lipid-structure incorporating SHEMS shows an acidity-sensitive release control behavior.

Experimental Example 4 Enhancement of Macromolecular Drug Encapsulation

In order to evaluate the degree of encapsulation of the macromolecular drug that can be supported on the inner phase of the lipid-based nanocarrier introduced with the SHEMS provided by the present invention, the liposomes of Example 4 and Example 9 were evaluated. To prepare a liposome dispersion containing BSA (Bovine Serum Albumin) in the interior, free BSA not embedded in the liposome was centrifuged at 1000 rcf for 30 minutes using a microcentricon YM-100 (MW 100KD) filter. After removal, the removed BSA solution was quantified by a BCA ^™ protein assay to determine the amount of BSA initially incorporated into the liposome structure by measuring the amount of BSA on the liposome trauma. In this way, the amount of BSA initially incorporated into the liposomes of the liposome dispersion prepared by the film hydration method using dioleoylphosphatidylethanolamine as the main acidity-sensitive lipid-structure as a main component. It was used as a control.

Lipid composition of the control b (FIG. 5B) liposomes is similar to the lipid-structure of Example 4, and the dioleoylphosphatidylethanolamine and CHEMS (cholesteryl hemisuccisinate; Korean patent application No. 2004- 117940), and also the lipid composition of the control d (FIG. 5D) liposomes was the same as the lipid-structure of Example 9 with phosphatidylcholine: dioleoylphosphatidylethanolamine (molar ratio 2: 3) and known acidity- It was prepared using CHEMS, a sensitivity stabilizer. The measured initial BSA amounts in Examples 4 and 9 and the corresponding control liposomes in FIG. 5 are shown in FIG. 5 in terms of their relative values using 100% of the amount of BSA used in preparation (a: Example 4, b: control, c: Example 9, d: control).

Lipid-based nanocarriers incorporating SHEMS provided in the present invention have relatively higher incidence of proteins in comparison with conventional control liposomes even when the phospholipid component composition ratios of the lipid-based transporters used in Examples 4 and 9 are different. As it was confirmed that it can be incorporated into, it was confirmed that the utility as various macromolecular drug carriers is greater than the existing liposomes.

Test Example 5 Efficacy Enhancement Effect of Biologically Active Components by Nanotransmitters

In order to evaluate the effect of enhancing the potency of the active ingredient appeared by increasing the transfer efficiency of the bioactive ingredient incorporated into the inner shell of lipid-based nanocarriers introduced SHEMS provided in the present invention, the whitening effect on the liposome inner shell of Example 4 Liposome dispersions containing 0.5 mM and 0.25 mM actinomycetes culture extracts were added to a cell culture medium [MEM) containing 1% (v / v) antibiotic and 10% serum bovine serum. , Human melanoma cell line (Human melanoma cell line) was applied to HM3KO culture, which was incubated for 5 days in a 37 ℃ humidified incubator supplied with 5% CO ₂ (Fig. 6b). The culture containing the liposomes was changed to a new one every two days. As a control group, the cells treated with the phosphate buffer solution (PBS) and the bioactive components not incorporated into the liposomes were added to the culture medium at concentrations of 1 mM, 0.5 mM, and 0.25 mM. Treated cells (Fig. 6A) were used.

Actinomycetes culture extract, a whitening agent, is an inhibitor that inhibits N-glycosylation, which is required in the pigment production step of tyrosinase, which plays an important role in producing melanin pigment in the body. Therefore, it has a whitening effect of reducing melanin production by inhibiting the normal activity of tyrosinase. In order to evaluate the effect of enhancing the efficacy, the absorbance at 490 nm was measured to determine the amount of melanin produced by HM3KO cells, and the culture medium to which the same amount of phosphate buffer solution (PBS) as the volume of the liposome dispersion treated in the experimental group was added. The treated cells were used as a positive control to measure the amount of melanin production, which was expressed based on the amount of melanin production 100%. Thus, the treatment results of the control group and Example 4 are shown in FIG. As shown in Figure 6, the lipid-based nano-carriers (Fig. 6b) in which the SHEMS of Example 4 is introduced (FIG. 6b) efficiently delivers the whitening components of 0.5mM and 0.25mM actinomycetes culture extracts into cells as compared to the control group (Fig. 6a). It was confirmed that the whitening effect to suppress the production of melanin.

In addition, after incubation for 5 days, the changed properties of the cells were observed before measuring the melanin production. The experimental group was treated with SHEMS-introduced lipid-based nanocarriers carrying the 0.5 mM actinomycetes culture extract of Example 4 (FIG. 7A) and the control group was prepared with the same composition as the control group b used in Test Example 4 as an existing acidity. -HM3KO cells (FIG. 7B) cultured by treating liposomes carrying 0.5 mM actinomycetes culture extract inside the sensitive lipid constructs were observed in an optical microscope, and are shown in FIG. As shown in FIG. 7A, the cells treated with the lipid-structure prepared by Example 4 were observed to have well developed dendrite, and compared with the acidity-sensitive liposome treatment control (FIG. 7B). Little effect on normal growth was observed.

Lipid-based nanocarriers incorporating SHEMS provided by the present invention can enhance the melanin production inhibitory effect by efficiently delivering the efficacy of the internally incorporated physiologically active ingredients within the cell without affecting the normal activity of the cells. there was.

Test Example 6 Verification of Intracellular Delivery Efficiency of Lipid-Based Nanocarriers Incorporating SHEMS

The intracellular delivery efficiency of the material encapsulated inside the acidity sensitive lipid-based nanocarrier in which the SHEMS provided by the present invention was introduced was quantitatively tested by analyzing with a flow activated cell sorter (FACS). In order to encapsulate the labeling substance in the lipid-based nanocarrier, a liposome dispersion prepared by preparing purified water containing the fluorescent substance calcein (calcein) and encapsulating calcein in the lipid-based nanocarrier of Example 4 was cultured. A certain amount was added to the HM3KO cells for 4 hours (FIG. 8B). Culture treated cells containing liposomes composed of lipid components of phosphatidylcholine: cholesterol (molar ratio 3: 2) carrying the same amount of calcein were used as a control (FIG. 8A). Cells treated for 4 hours were washed three times with PBS, and cells implanted with trypsin / EDTA were dropped from the bottom of the culture vessel and centrifuged for 5 minutes at 1000 rpm. Centrifuged cell pellet was resuspended in PBS and analyzed by FACS.

 Intracellular delivery of calcein (calcein, Ex / Em = 494/517 nm) enclosed inside the acidity sensitive lipid-based nanocarriers into which the SHEMS of Example 4 was introduced was assessed by FACS. In terms of its properties, calceindo, a substance that cannot penetrate the cell membrane, was encapsulated in acid-sensitive lipid-based nanocarriers into which SHEMS was introduced, and the amount delivered to cells was estimated to be greater than that enclosed in control liposomes. In conclusion, FACS analysis showed that acidity sensitive lipid-based nanocarriers incorporating SHEMS are more efficient in intracellular delivery than lipid-based constructs that are not acidity sensitive.

<Formulation Examples 1-16> Cream Formulation

Cream formulations containing the liposome dispersions prepared in Examples 1-9 and the nanoemulsions prepared in Examples 10-16 were prepared according to conventional preparation methods with the compositions shown in Tables 4 and 5, respectively.

Liposomal-Containing Cream Formulations ingredient Example of formulation (% by weight) One 2 3 4 5 6 7 8 9 Glyceryl Stearate 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Polysorbate 60 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Cetostearate 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 Wax 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Squalane 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Sorbitan sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cetylethylhexanoate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Liquid paraffin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Active ingredient mixture 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 glycerin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Example 1 5.0 - - - - - - - - Example 2 - 5.0 - - - - - - - Example 3 - - 5.0 - - - - - - Example 4 - - - 5.0 - - - - - Example 5 - - - - 5.0 - - - - Example 6 - - - - - 5.0 - - - Example 7 - - - - - - 5.0 - - Example 8 - - - - - - - 5.0 - Example 9 - - - - - - - - 5.0 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance

Nanoemulsion-containing cream formulation ingredient Example of formulation (% by weight) 10 11 12 13 14 15 16 Glyceryl Stearate 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Polysorbate 60 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Cetostearate 2.5 2.5 2.5 2.5 2.5 2.5 2.5 Wax 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Squalane 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Solbitan Sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cetylethylhexanoate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Liquid paraffin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Active ingredient mixture 5.0 5.0 5.0 5.0 5.0 5.0 5.0 glycerin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Example 10 10.0 - - - - - - Example 11 - 10.0 - - - - - Example 12 - - 10.0 - - - - Example 13 - - - 10.0 - - - Example 14 - - - - 10.0 - - Example 15 - - - - - 10.0 - Example 16 - - - - - - 10.0 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance

<Formulation Examples 17-32> Softening Longevity Formulation

The flexible longevity formulations containing the liposome dispersions prepared in Examples 1-9 and the nanoemulsions prepared in Examples 10-16 were prepared according to conventional preparation methods with the compositions shown in Tables 6 and 7, respectively.

Liposomal Containing Softener Formulation ingredient Example of formulation (% by weight) 17 18 19 20 21 22 23 24 25 Betaine 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Natto gum 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Cellulose gum 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 ethanol 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Polyoxyethylene Cured Castor Oil 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Tocopherol Acetate 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Example 1 5.0 - - - - - - - - Example 2 - 5.0 - - - - - - - Example 3 - - 5.0 - - - - - - Example 4 - - - 5.0 - - - - - Example 5 - - - - 5.0 - - - - Example 6 - - - - - 5.0 - - - Example 7 - - - - - - 5.0 - - Example 8 - - - - - - - 5.0 - Example 9 - - - - - - - - 5.0 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance

Nanoemulsion Containing Softener Formulation ingredient Example of formulation (% by weight) 26 27 28 29 30 31 32 Betaine 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Natto gum 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Cellulose gum 0.005 0.005 0.005 0.005 0.005 0.005 0.005 ethanol 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Polyoxyethylene Cured Castor Oil 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Tocopherol Acetate 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Example 10 7.0 - - - - - - Example 11 - 7.0 - - - - - Example 12 - - 7.0 - - - - Example 13 - - - 7.0 - - - Example 14 - - - - 7.0 - - Example 15 - - - - - 7.0 - Example 16 - - - - - - 7.0 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance

<Formulation examples 33 to 48> nutrient cosmetics formulation

The nutrient longevity formulations containing the liposome dispersions prepared in Examples 1 to 9 and the nanoemulsions prepared in Examples 10 to 16 were prepared according to conventional preparation methods with the compositions shown in Tables 8 and 9, respectively.

Liposomal-containing nutrient formulation ingredient Example of formulation (% by weight) 33 34 35 36 37 38 39 40 41 Cetylethylenehexanoate 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Lipophilic Monostearylic Acid Stearate 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Squalane 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Example 1 5.0 - - - - - - - - Example 2 - 5.0 - - - - - - - Example 3 - - 5.0 - - - - - - Example 4 - - - 5.0 - - - - - Example 5 - - - - 5.0 - - - - Example 6 - - - - - 5.0 - - - Example 7 - - - - - - 5.0 - - Example 8 - - - - - - - 5.0 - Example 9 - - - - - - - - 5.0 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Sorbitan sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Triethanolamine 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Carboxyvinyl polymer 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance

 Nutritional Longevity Formulation with Nanoemulsion ingredient Example of formulation (% by weight) 42 44 44 45 46 47 48 Cetylethylenehexanoate 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Lipophilic Monostearylic Acid Stearate 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Squalane 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Example 10 8.0 - - - - - - Example 11 - 8.0 - - - - - Example 12 - - 8.0 - - - - Example 13 - - - 8.0 - - - Example 14 - - - - 8.0 - - Example 15 - - - - - 8.0 - Example 16 - - - - - - 8.0 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Sorbitan sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Triethanolamine 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Carboxyvinyl polymer 0.2 0.2 0.2 0.2 0.2 0.2 0.2 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance

<Formulation Examples 49-64> Essence Formulation

The essence formulations containing the liposome dispersions prepared in Examples 1-9 and the nanoemulsions prepared in Examples 10-16 were prepared according to conventional preparation methods with the compositions shown in Tables 10 and 11, respectively.

Liposomal Containing Essence Formulation ingredient Example of formulation (% by weight) 49 50 51 52 53 54 55 56 57 Glyceryl Stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Solbitan Sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Cetylethylhexanoate 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Squalane 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 lanolin 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 glycerin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Carboxyvinyl polymer 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Hydrolyzed Collagen 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Triethanolamine 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Example 1 5.0 - - - - - - - - Example 2 - 5.0 - - - - - - - Example 3 - - 5.0 - - - - - - Example 4 - - - 5.0 - - - - - Example 5 - - - - 5.0 - - - - Example 6 - - - - - 5.0 - - - Example 7 - - - - - - 5.0 - - Example 8 - - - - - - - 5.0 - Example 9 - - - - - - - - 5.0 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance

Nano emulsion-containing essence formulation ingredient Example of formulation (% by weight) 58 59 60 61 62 63 64 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Squalane 7.0 7.0 7.0 7.0 7.0 7.0 7.0 Polysorbate 60 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Sorbitan monostearate 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Example 10 10.0 - - - - - - Example 11 - 10.0 - - - - - Example 12 - - 10.0 - - - - Example 13 - - - 10.0 - - - Example 14 - - - - 10.0 - - Example 15 - - - - - 10.0 - Example 16 - - - - - - 10.0 glycerin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Triethanolamine (TEA) 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Carboxyvinyl polymer 0.2 0.2 0.2 0.2 0.2 0.2 0.2 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance Balance

As discussed above, lipid-based nanocarriers such as stigmas-5,22-diene-3-ol, hydrolysis butanedioate (SHEMS), or liposomes or nanoemulsions, according to the present invention are conventionally acid-sensitive. Compared to liposomes, relatively large amounts of physiologically active ingredients could be loaded into the internal organs, and they reacted specifically in the general acidity range 6.0-4.5 of endosomes, which are organelles in living cells, thereby degrading and degrading lipid membrane stability. It was possible to effectively release the physiologically active ingredient in the cell to enhance the effect as a carrier. Therefore, it is excellent in carrying capacity of bioactive components having whitening enhancement and / or anti-wrinkle and anti-aging and treatment effects, and can function as a transporter capable of doubling the physiological activity in the body, and has a small particle size and colloidal stability. It has excellent chemical stability and can be used in various external skin preparations. In addition, the phospholipid construct has an advantage that is harmless to the human body can be usefully used to enhance the bioavailability of the pharmaceutical composition or health food composition for oral administration.

Claims (12)

As lipid-based nanocarriers, A lipid structure prepared by mixing a phospholipid component and stigmasteryl hemisuccinate represented by Chemical Formula 1 below; And Consists of a structure in which a bioactive component is embedded in the lipid structure Lipid-based nanocarriers prepared to disintegrate below pH 6 to release bioactive components: [Formula 1]

The lipid-based nanocarrier of claim 1, wherein the nanocarrier has an average particle diameter of 10 nm to 1,000 nm. The method of claim 1, wherein the stigmasteryl hemisuccinate is selected from the group consisting of stigmasteryl hemisuccinate; Or one selected from tris salt, tris (hydroxymethyl) aminomethane salt and cyclohexyl ammonium salt thereof; Or a mixture of at least one selected from stigmasteryl hemisuccinate and salts thereof. According to claim 1, The phospholipid component of the lipid structure is egg yolk lecithin (phosphatidylcholine) having a fatty acid chain having 12 to 24 carbon atoms, soybean lecithin, risorecithin, sphingomyelin, phosphatidic acid, phosphatidylserine, phosphatidylglycerol, Phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, plasmagen and hydrogenated products of said phospholipids; Dicetylophosphate, distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostaroylphosphatidylcholine, eleostaloylolamine, Osteoaroylphosphatidylserine; And a fatty acid mixture obtained by hydrolyzing them; at least one selected from the group consisting of lipid-based nanocarriers. 5. The lipid-based nanocarrier according to claim 4, wherein the mixing ratio of the maximum component to the minimum component is 1: 5 or less when two or more kinds of the above-described lipid components are mixed. The lipid-based nanocarrier of claim 1, wherein the lipid component: stigmasteryl hemisuccinate is mixed at a molar ratio of 9: 1 to 1: 9. The lipid-based nanocarrier of claim 1, wherein the lipid component including the stigmasteryl hemisuccinate is used in an amount of 0.001 to 20% by weight based on the total weight of the nanocarrier. The lipid-based nanocarrier according to claim 1, wherein the bioactive component is a component for enhancing whitening or for preventing wrinkles and aging. The lipid-based nanocarrier of claim 8, wherein the bioactive component is incorporated in an amount of 0.01 to 30 wt% based on the total weight of the nanocarrier. delete A composition for external application for skin, comprising the lipid-based nanocarrier according to any one of claims 1 to 9. 12. The composition of claim 11, wherein the composition comprises softening cream, astringent makeup, nutrition cream, nutrition cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, may Cup base, foundation, body lotion, body cream, body oil, body essence, body cleanser, hair dye, shampoo, rinse, toothpaste, mouthwash, hair dresser, wool, lotion, ointment, gel, cream, patch and spray The external preparation composition for skin, characterized in that it has a formulation selected from.

Images