JP5728152B2 - Dry liposome preparation - Google Patents

Description

  The present invention relates to a technique for a dry liposome preparation in which retinol palmitate and astaxanthin are stably blended.

Liposomes containing retinoids or carotenoids for the purpose of reducing irritation are known (Patent Document 1: JP-A-62-215513). It is advantageous to add a lipophilic antioxidant such as α-tocopherol to the lipid layer, and add a water-soluble antioxidant such as ascorbic acid to the aqueous phase. It can be stored for a long time without doing so. However, the stability of retinoids or carotenoids has not been evaluated.
The retinol palmitate used in the present invention has been used in skin external preparations such as creams and emulsions for the purpose of acne treatment and wrinkle improvement. Retinol palmitate is more stable than retinol, but has the disadvantage of being altered by oxygen and ultraviolet light. There is a demand for a preparation technique in which retinol palmitate is stabilized.
In order to mix water-immiscible carotenoids with water, liposomes filled with carotenoids are known (Patent Document 2: JP-T-2004-518707). Astaxanthin is exemplified as a carotenoid, and lyophilization of liposomes is also described. However, the stability of carotenoids has not been evaluated.
A composition containing an astaxanthin and an antioxidant is known (Patent Document 3: Japanese Patent Laid-Open No. 9-143063). Examples of antioxidants include vitamin A and vitamin C. However, the stability of astaxanthins and antioxidants has not been evaluated.
Astaxanthin is excellent in anti-aging effect, anti-inflammatory effect and anti-oxidation effect, and is used as a skin external preparation. However, astaxanthin has the disadvantage that it is easily decomposed by heat, light, oxygen, etc. and has poor stability during storage, and there is a need for a formulation technique for stably incorporating astaxanthin.
Addition of ascorbic acid to astaxanthin improves the storage stability of astaxanthin (Patent Document 4: Japanese Patent Laid-Open No. 6-264055), and seals astaxanthin together with an oxygen scavenger to improve the storage stability of astaxanthin (Patent Document 5: JP-A-8-12896) is known.
The liposome preparation produced by spray-drying an aqueous dispersion of liposomes is very unstable, and in order to improve stability, an aqueous dispersion of liposomes containing saccharides and having an ion concentration of about 20 mM or less is sprayed. A liposome preparation produced by drying is known (Patent Document 6: Japanese Patent Application Laid-Open No. 64-3115).

JP-A-62-215513 Special table 2004-518707 gazette Japanese Patent Laid-Open No. 9-143063 JP-A-6-264055 JP-A-8-12896 JP-A 64-3115

  It is to provide a dry liposome preparation in which retinol palmitate and astaxanthin are stably blended.

The main configuration of the present invention is as follows.
(1) A dry liposome preparation containing retinol palmitate, astaxanthin, ascorbic acid and / or a salt thereof.
(2) The drying according to (1), wherein the amount of astaxanthin is 1 to 10 parts by weight and the amount of ascorbic acid is 40 to 200 parts by weight when the amount of retinol palmitate is 100 parts by weight. Liposome preparation.
(3) The dry liposome preparation according to (1) or (2), further comprising sucrose.
(4) The dry liposome preparation according to any one of (1) to (3), which is prepared by freeze-drying or spray-drying an ascorbic acid aqueous solution in which liposomes containing retinol palmitate and astaxanthin are dispersed.
(5) The dry liposome preparation according to any one of (1) to (4) is used as the first agent, the aqueous preparation is used as the second agent, and the first agent and the second agent are mixed at the time of use. An external preparation for skin characterized by
(6) The dry liposome preparation according to (1), wherein the amount of astaxanthin is 3 parts by weight and the amount of ascorbic acid is 70 parts by weight when the amount of retinol palmitate is 100 parts by weight.

Dry liposome preparations obtained by drying liposome dispersions containing retinol palmitate, astaxanthin and ascorbic acid have improved storage stability, and when re-dissolved, degradation of retinol palmitate and astaxanthin is suppressed and reproducibility. Is expensive. When the blending amount of retinol palmitate is 100 parts by weight, it is suitable that the blending amount of astaxanthin is 1 to 10 parts by weight and the blending amount of ascorbic acid is 40 to 200 parts by weight. The addition of sucrose can contribute to the protection of liposomes from lyophilization.
The dry liposome preparation of the present invention can be obtained by freeze-drying or spray-drying an ascorbic acid aqueous solution in which liposomes containing retinol palmitate and astaxanthin are dispersed.
The dry liposome preparation of the present invention can be applied to the skin by applying retinol palmitate and astaxanthin close to the initial content to the skin by being dispersed in an aqueous solution or emulsion at the time of use.

  The retinol palmitate used in the present invention is a retinol palmitate ester. Retinol palmitate is a pale yellow to yellow-red solid or oily substance with a melting point of 28-29 ° C. It is oil-soluble and hardly soluble in water. A commercial item can be used for retinol palmitate. For example, DSM Nutrition Japan Co., Ltd. product: Vitamin A palmitate, Riken Vitamin Co., Ltd. product: RIKEN A Palmitate 1000 (E) can be mentioned.

  Astaxanthin used in the present invention is a carotenoid discovered from crayfish eggs, and is present in crustaceans such as crabs and shrimps, salmon meat, and kingfisher epidermis. It is a glossy purple-red plate crystal and has a melting point of 216 ° C. Astaxanthin can use a commercial item. For example, Fuji Chemical Industry Co., Ltd. product: Asterel oil 50F is mentioned.

  Ascorbic acid used in the present invention is a substance also called vitamin C, and is involved in redox action in vivo. Ascorbic acid is frequently used as a water-soluble antioxidant. Ascorbic acid is a single swash plate or needle crystal and has a melting point of 190 to 192 ° C. Ascorbic acid used in the present invention can be used as a salt such as an alkali metal salt. A commercial item can be used for ascorbic acid and / or its salt. For example, BASF Japan Co., Ltd. product: Japanese Pharmacopoeia Ascorbic acid 100 mesh, DSM Nutrition Japan Co., Ltd. product: L-sodium ascorbate crystal (food additive) is mentioned.

The dry liposome preparation of the present invention means a preparation prepared by preparing an aqueous dispersion of liposomes and drying it by a technique such as freeze drying or spray drying.
The aqueous dispersion of liposome can be produced by the following method. One example will be explained in detail. The liposome membrane component substance, retinol palmitate and astaxanthin are dissolved in ethanol, and an aqueous solution in which ascorbic acid, sucrose, pH adjuster, etc. are dissolved is added, and the mixture is heated to 65 ° C. Emulsify with. This emulsified suspension can be filtered under pressure to produce an aqueous dispersion of liposomes.
0.01 to 0.1 parts by weight of retinol palmitate, 0.0005 to 0.005 parts by weight of astaxanthin, and 0.02 to 0.1 parts by weight of ascorbic acid are added to 1 part by weight of the membrane component substance. It is preferable. When preparing an aqueous dispersion of liposomes, it is preferable to suspend the membrane component lipid in a proportion of 0.001 to 0.05 parts by weight with respect to 1 part by weight of water.

  As for the above membrane component substances, in addition to phospholipids typified by phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, lysophosphatidylcholine, sphingomyelin, egg yolk lecithin, soybean lecithin, etc., glycolipid, a kind of dialkyl type surfactant The above mixture is the main component. Then, sterols such as cholesterol and cholestanol as membrane stabilizers, dicetyl phosphate, phosphatidic acid, ganglioside, stearylamine and the like as charged substances, and α-tocopherol and the like as antioxidants are added to the membrane components. A substance may be formed.

  The liposome aqueous dispersion thus obtained can be made into a dry liposome preparation by freeze drying, spray drying or the like. For example, as a freeze-drying method, a normal freeze-drying operation may be performed after the above-mentioned aqueous dispersion of liposomes is filled into containers such as vials. Desirable operations in lyophilization include rapid freezing at a freezing temperature of -20 to -80 ° C, preferably -60 ° C, and sublimation of water under reduced pressure conditions of 10 Pa (0.075 Torr) or less. . Further, as a spray drying method, after removing the solvent from the liposome aqueous dispersion by spray drying, the powder can be aseptically divided and sealed. As a desirable operation in spray drying, the inlet temperature is set to 110 to 200. C., preferably 120.degree. C. to 150.degree. C. can be mentioned.

  The dry liposome preparation thus obtained has a stabilized liposome structure and retinol palmitate and astaxanthin, so that the effect of retinol palmitate and astaxanthin is effective when dispersed in an aqueous preparation at the time of use. Can be demonstrated. In particular, regarding the storage stability of retinol palmitate and astaxanthin, the stability was significantly improved by the synergistic effect of retinol palmitate, astaxanthin, and ascorbic acid (salt).

  The aqueous preparation in which the dry liposome preparation is dispersed is an aqueous solution, an emulsified composition in which the outer phase is an aqueous solution, or a dispersion composition in which the outer phase is an aqueous solution. For example, lotion, O / W emulsion, O / W cream, aqueous Gels, serums, packs and the like can be mentioned. In addition, the dry liposome preparation of the present invention is premixed and stored in a powder raw material not containing water, for example, sugars such as maltose and sucrose, amino acids, vitamin C and the like, and dispersed in an aqueous preparation at the time of use. Can also be used. The dry liposome preparation of the present invention or the liposome dispersion prepared by dispersing the dry liposome preparation of the present invention in an aqueous preparation can be used as a skin external preparation for cosmetics, quasi drugs, pharmaceuticals and the like.

Preparation of liposome dispersion Components 1 to 4 were mixed and dissolved in ethanol at the composition shown in Table 1. Upon dissolution, it was heated to 65 ° C. and confirmed to be completely dissolved. As for 30% retinol palmitate, Riken Vitamin Co., Ltd .: RIKEN A Palmitate 1000 (E) was used. RIKEN A Palmitate 1000 (E) contains 30% retinol palmitate. As 5% astaxanthin oil, Fuji Chemical Industry Co., Ltd. product: Asteril oil 50F was used. 5% astaxanthin oil contains 5% (w / w) astaxanthin.
Next, an aqueous solution in which the components 6 to 9 in Table 1 were dissolved was added to the solution, and the mixture was dispersed by stirring at 10,000 rpm for 15 minutes with a homomixer while heating at 65 ° C. Then, it filtered under the high pressure with the polycarbonate membrane filter, and prepared the liposome dispersion liquid for Example 1 and the liposome dispersion liquid for Comparative Examples 1-8.

Preparation of dry liposome preparation The liposome dispersion for Example 1 obtained and the liposome dispersion for Comparative Examples 1 to 8 were filled into vials in 1.75 mL portions, and after half-plugging, the shelf temperature was cooled to -60 ° C in advance. The sample was lyophilized under normal lyophilization conditions. After drying, it was purged with nitrogen and capped to obtain dry liposome preparations of Example 1 and Comparative Examples 1-8. The composition (% by weight) of the dry liposome preparation is shown in Table 2. The blending ratio of retinol palmitate, astaxanthin and ascorbic acid in Example 1 is 100: 3: 70.

Evaluation of Storage Stability The dry liposome preparations of Example 1 and Comparative Examples 1 to 8 were stored at 40 ° C. for 3 months, and each 3 dry liposome preparations were condensated with 5.0 mL of water per vial every month. Then, the concentration, particle size and pH of retinol palmitate and astaxanthin were measured.
For measuring the concentrations of retinol palmitate and astaxanthin, a solution obtained by subjecting each condensate dispersion to 20-fold dilution with ethanol exactly 0.45 μm non-aqueous filter was used as a sample. When measuring the retinol palmitate concentration, a certain amount of an internal standard solution (dl-α-tocopherol acetate / ethanol solution or steryl glycyrrhetinate / ethanol solution) was accurately added in advance at the time of dilution.
Each standard solution is prepared at the time of use. In the case of retinol palmitate, each concentration is calculated from the area ratio of the standard solution to the internal standard, and in the case of astaxanthin, the concentration is calculated from the absorbance of the standard solution. %).

The analysis conditions for retinol palmitate are shown below.
Liquid chromatography analysis conditions
Analytical column: YMC-Pack, ODS-AQ (AQ-302) 4.6mm Φ x 150mm
Column bath temperature: 40 ° C
Eluent: Methanol: Water = 95: 5
Eluent flow rate: 1.0 mL / min Sample injection volume: 5 μL
Detector: UV 280nm

The analysis conditions for astaxanthin are shown below.
Absorbance analysis conditions
Measurement wavelength: 478.0nm
Control cell: ethanol

For the measurement of particle diameter, a solution obtained by diluting each condensate dispersion 20 times with water is used as a sample, and NICOMP 380ZLS (Particle Sizing Systems) is used for the dynamic light scattering method (volume weighted Gaussian distribution method). The average particle size was measured.
The pH was measured directly on each condensate dispersion.

  The evaluation of color stability was based on the product stored at 5 ° C for 3 months, and the change in color of the product stored at 40 ° C for 3 months was evaluated. A sample having no change in color and having no problem with color stability was evaluated as “◯”, a product having a slight change in color was evaluated as “Δ”, and a sample having a change in color and having a problem in stability was evaluated as “X”.

  Table 3 shows the composition (% by weight) after condensing and the storage stability evaluation results of the dry liposome preparation.

Retinol palmitate and astaxanthin are extremely unstable when incorporated into lyophilized liposomes alone. As shown in Comparative Example 1, in the freeze-dried liposome containing retinoyl palmitate alone, the concentration of retinol palmitate is reduced to 56.6% after storage at 40 ° C. for 3 months. Moreover, as shown in Comparative Example 2, in the lyophilized liposome containing astaxanthin alone, the concentration of astaxanthin decreases to 85.4% after storage at 40 ° C. for 3 months.
Then, when retinol palmitate, which is said to be excellent in wrinkle improvement, and astaxanthin, which is said to be excellent in anti-aging effect, were simultaneously blended into a freeze-dried liposome preparation (Comparative Example 3), the retinol palmitate was stored at 40 ° C. for 3 months. The residual amount was 67.2%, which was improved in stability as compared with the one containing retinol palmitate alone (56.6%). Astaxanthin had a residual amount of 69.degree. It was 3%, and the stability was greatly reduced as compared with the compound containing astaxanthin alone (85.4%). When retinol palmitate and astaxanthin are each incorporated into a freeze-dried liposome, there is a problem that stability is low. However, when they coexist, there is a problem that the stability of astaxanthin is lowered.

When each of retinol palmitate and astaxanthin and ascorbic acid are added to a freeze-dried liposome, the stability of retinol palmitate and astaxanthin is improved. As shown in Comparative Example 4, when ascorbic acid is allowed to coexist with retinol palmitate, the residual amount of retinol palmitate after storage at 40 ° C. for 3 months is 85.5%, which improves stability. However, there was a problem that the color changed when stored at 40 ° C for 3 months. As shown in Comparative Example 5, when ascorbic acid coexists with astaxanthin, the residual amount of astaxanthin after storage at 40 ° C. for 3 months is significantly improved to 93.2%.
In Comparative Example 3 in which retinol palmitate and astaxanthin coexisted, there was a problem that the stability of astaxanthin was lowered. It was found that a lyophilized liposome preparation with significantly improved stability was obtained for both astaxanthin (Example 1). As shown in Example 1, the residual amount of retinol palmitate after storage at 40 ° C. for 3 months was 90.2%, and the residual amount of astaxanthin was 94.4%, indicating extremely high stability. Contrary to the expectation that the stability of astaxanthin of Example 1 would decrease from the knowledge of Comparative Example 3, the stability of astaxanthin was stored at 40 ° C. for 3 months when the astaxanthin of Comparative Example 5 and ascorbic acid were blended. In Example 1, the residual amount of astaxanthin was improved to 94.4% by further incorporating retinol palmitate from the residual amount of astaxanthin of 93.2%. Further, with respect to retinol palmitate, the retinol palmitate remaining amount of 85.5% after storage at 40 ° C. for 3 months when the retinol palmitate and ascorbic acid of Comparative Example 4 were blended was 90.2% in Example 1. Improved. And the freeze-dried liposome of Example 1 is excellent in color stability.

  The effect of α-tocopherol, known as a stabilizer, was also examined. In Comparative Example 6, retinol palmitate and α-tocopherol were added to lyophilized liposomes. The residual amount of Retinol palmitate of Comparative Example 6 after storage at 40 ° C. for 3 months is 60.8%, which is more stable than Comparative Example 1 (Retinol palmitate remaining amount of 56.6%) blended solely with retinol palmitate. Improved. However, in Comparative Example 7 in which retinol palmitate, ascorbic acid, and α-tocopherol were blended in freeze-dried liposomes, the residual amount of retinol palmitate stored at 40 ° C. for 3 months was 83.1%, and retinol palmitate and ascorbic acid As compared with the residual amount of retinol palmitate of Comparative Example 4 containing 85.5%, the stability was lowered. α-Tocopherol inhibits the stabilizing effect of ascorbic acid retinol palmitate.

[Prescription Example 1]
Lyophilized liposome containing retinol palmitate, astaxanthin and ascorbic acid prepared in Example 1 of the first agent 0.09 g
2nd Agent Lotion of the following composition 0.91g
Ingredient Mass%
Glycerin 2.0
1,2-pentanediol 5.0
Trimethylglycine 0.5
Sodium hyaluronate 0.0001
Sodium citrate appropriate amount Citric acid appropriate amount Purified water Residual
Usage The first agent is dissolved in the second agent at the time of use.

[Prescription Example 2]
First lyophilized liposome containing retinol palmitate, astaxanthin and ascorbic acid prepared in Example 1 0.03 g
Emulsions of the second agent composition shown below 0.97g
Ingredient Mass%
Glycerin 5.0
Dipropylene glycol 10.0
Sodium hyaluronate 0.0001
Xanthan gum 0.1
Squalane 5.0
Dimethicone 1.0
Jojoba oil 1.0
Lauroyl glutamate di (phytosteryl / octyldodecyl) 1.0
Sorbitan stearate 0.2
Palm oil fatty acid sucrose 0.1
PEG-60 hydrogenated castor oil 0.5
Carbomer K 0.05
Purified water residue
Usage The first agent is dissolved in the second agent at the time of use.

[Prescription Example 3]
Lyophilized liposome containing retinol palmitate, astaxanthin and ascorbic acid prepared in Example 1 of the first agent 0.09 g
Essence of the second agent composition shown below 0.91g
Ingredient Mass%
Glycerin 8.0
1,3-butylene glycol 5.0
Dipropylene glycol 5.0
Sodium hyaluronate 0.001
Sclerotium gum 0.01
Dimethicone 1.0
Macadamia nut oil 0.5
Macadamia nut fatty acid phytosteryl 0.5
Hydrogenated lecithin 0.5
Polyglyceryl stearate-10 0.5
Ethanol 5.0
Purified water residue
Usage The first agent is dissolved in the second agent at the time of use.

Claims (5)

  A dry liposome preparation containing retinol palmitate, astaxanthin, ascorbic acid and / or a salt thereof.   The dry liposome preparation according to claim 1, wherein the amount of astaxanthin is 1 to 10 parts by weight and the amount of ascorbic acid is 40 to 200 parts by weight, when the amount of retinol palmitate is 100 parts by weight.   Furthermore, sucrose is contained, The dry liposome formulation of Claim 1 or 2 characterized by the above-mentioned.   The dry liposome preparation according to any one of claims 1 to 3, which is prepared by freeze-drying or spray-drying an ascorbic acid aqueous solution in which liposomes containing retinol palmitate and astaxanthin are dispersed. The dry liposome preparation according to any one of claims 1 to 4 is a first agent, an aqueous preparation is a second agent, and the first agent and the second agent are mixed at the time of use to form a liposome dispersion, Skin external preparation.