JP2019069909A - Polyglycerol-containing liposome preparation - Google Patents

Abstract

To provide liposome preparations capable of allowing an active ingredient that is formulated in medicines, quasi-drugs, cosmetics, and the like and causes healing or improvement on human health to penetrate into a body through a surface, which is a boundary with the external world of a human body, particularly a skin, using a liposome that consists of a phospholipid having a hydrophilic group and a lipophilic group, and the like and is organized as a monolayer or multilayer vesicle, so as to store much more.SOLUTION: Provided is a liposome preparation for medicines or cosmetics which contains: (A) a polyglycerol with an average degree of polymerization of 2 to 20 calculated from a hydroxyl value; (B) at least one phospholipid ingredient, and (C) at least one active ingredient in a medicine or a cosmetic.SELECTED DRAWING: Figure 1

Description

  The present invention is a liposome that supplies active ingredients contained in medicines, quasi-drugs, cosmetics, etc. that brings about healing or improvement on human health through the surface that is the boundary with the external environment such as human mucous membrane or skin into the human body. It relates to a preparation.

  In the past, active ingredients that cure or improve the health of the human body are blended with other compounds and formulated or marketed as medicines, quasi-drugs, cosmetics etc. with the human body such as mucous membranes or skin, etc. It is regarded as important how it can be delivered to the body through the boundary surface.

  In particular, the skin of the human body is roughly classified into the epidermis that is the boundary with the external world, and the dermis located inside the epidermis, and is exposed to bacteria, fungi, viruses, etc. that are routinely present on the external world or the surface of the epidermis. It is equipped with a defense system that has a barrier function to prevent the intrusion of foreign substances that cause harm to the human body, and even if it is an active ingredient useful for the human body as described above, It is not easy to penetrate the dermis.

  Therefore, in order to allow the above-mentioned active ingredient to penetrate into the dermis through the epidermis, a method using a liposome composed of a phosphate lipid or the like having a hydrophilic group and a lipophilic group and organized as a unilamellar or multilamellar vesicle. It has been known. Liposomes can be microparticulated and have a structure similar to that of human cell membranes, so they are easily adaptable to the human body, and are effective means for causing the above-mentioned active ingredients to penetrate into the dermis through the epidermis.

  For example, Patent Document 1 discloses a liposome preparation comprising a predetermined amount of glycerin, a predetermined amount of a phospholipid component such as lecithin, and a pharmaceutically or cosmetically effective component, which is harmless and has an irritant action such as ethanol. It has been described that the effect of not exerting

Toko 2012-520245

  However, in the liposome preparation disclosed in Patent Document 1, the incorporation of glycerin improves the flexibility of the formed liposome, and as a result, the penetration and transport of the active ingredient into the deep skin layer. Although it is stated that the effect is enhanced, the liposome formulated with glycerin can not sufficiently retain the active ingredient which is incorporated therein to bring about healing or improvement on human health through the epidermis of the skin in the dermis, and they are effective. Further improvements have been sought to penetrate and transport the ingredients to the dermis for storage.

  Therefore, in the health of the human body, it is incorporated in medicines, quasi-drugs, cosmetics, etc. using a liposome composed of a phosphate lipid having a hydrophilic group and a lipophilic group and organized as unilamellar vesicles or multilamellar vesicles. It is an object of the present invention to provide a liposome preparation which can penetrate the body through the surface which is the boundary with the external environment of the human body, in particular, the skin, and retain more, as the active ingredient that causes healing or improvement.

  [1] That is, according to the present invention, (A) polyglycerin having an average degree of polymerization of 2 to 20 calculated from a hydroxyl value, (B) at least one phospholipid component, and (C) at least one pharmaceutical agent Or a pharmaceutical or cosmetic liposomal preparation characterized by containing an active ingredient in cosmetics.

  [2] And, the above-mentioned phospholipid component is at least one selected from phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl serine, phosphatidyl glycerol, diphosphatidyl glycerol, sphingomyelin, It is a liposome formulation as described.

  [3] And the said active ingredient is a vitamin and its derivative, an anti-inflammatory agent, an antiviral agent, an antibacterial agent, an antifungal agent, an antifungal agent, an analgesic agent, an antipruritic agent, an antioxidant, a moisturizing agent, a whitening agent, a hair restorer, an anti-wrinkle agent The liposome preparation according to the above [1] or [2], which is at least one selected from an agent and a cell activator.

  According to the present invention, it is formulated into pharmaceuticals, quasi-drugs, cosmetics, etc., using a liposome composed of a phosphate lipid having a hydrophilic group and a lipophilic group and organized as a unilamellar or multilamellar vesicle. The active ingredient that causes healing or improvement on human health can penetrate into the body through the surface that is the boundary with the external body of the human body, especially the skin, and can be stored more.

It is a figure which shows visually the accumulation amount of ascorbic acid cetyl ether (VC-cetyl ether) in epidermis and dermis. It is a figure which shows visually the accumulation amount of tocopherol acetate (Tocopherol acetate) in epidermis and dermis.

  Hereinafter, embodiments of the liposome preparation according to the present invention will be described in detail. Note that although the embodiments described below are specific examples preferable for practicing the present invention, various technical limitations have been made, but the present invention is intended to particularly limit the invention in the following description. It is not limited to this form unless otherwise specified. Further, a notation expressing a numerical range includes an upper limit and a lower limit.

  The liposome preparation of the present invention contains a predetermined amount of polyglycerin, a predetermined amount of at least one phospholipid component, and at least one active ingredient in medicine or cosmetics.

  The polyglycerol having an average degree of polymerization of 2 to 20 calculated from the hydroxyl value of the component (A) of the present invention is a polyhydric alcohol obtained by dehydration condensation reaction, addition reaction or the like using glycerin or epichlorohydrin as a raw material. And as polyglycerin of the average degree of polymerization 2-20 computed from the hydroxyl value, diglycerin, triglycerin, tetraglycerin, pentaglycerin, hexaglycerin, nonaglycerin, decaglycerin etc. are preferable, and also the said average degree of polymerization 2 More preferably, it is 15 polyglycerin. By using polyglycerin having an average degree of polymerization calculated from the hydroxyl value in the above range, it is incorporated into medicines, quasi drugs, cosmetics, etc., and penetrates to the dermis of the active ingredient that causes healing or improvement in human health. It can be stored. Moreover, the said polyglycerol can use 1 type, or 2 or more types of those. Furthermore, in order to use the liposome preparation of the present invention for the human body, the above-mentioned polyglycerin obtained by dehydration condensation using glycerin having high safety for human body is used without using epichlorohydrin which is toxic for human body. Is preferred.

The average degree of polymerization of the above polyglycerin is calculated from the hydroxyl value and is calculated by the formula (i) shown below. Moreover, the hydroxyl value in Formula (i) is measured based on "the reference | standard fats-and-oils analysis test method" (Japanese Yeast Chemical Society establishment). Specifically, when 1 g of a sample is acetylated with an acetic anhydride / pyridine solution, it is represented by the number of mg of potassium hydroxide (KOH) required to neutralize acetic acid bonded to a hydroxyl group, and the following formula (ii) It is determined by
Average degree of polymerization = (112.2 x 103-18 x hydroxyl number) / (74 x hydroxyl number-56.1 x 103) (i)
Hydroxyl value = (ab) x 28.05 / amount of sample collected (g) ... (ii)
a: Consumption of 0.5 N potassium hydroxide solution by blank test (ml)
b: Consumption of 0.5 N potassium hydroxide solution according to this test (ml)

  As polyglycerol having an average degree of polymerization of 2 to 20 calculated from the hydroxyl value of the component (A), those synthesized as described above can be used, but commercial products can also be used. As commercially available products, for example, diglycerin S manufactured by Sakamoto Yakuhin Kogyo Co., Ltd., polyglycerin # 310, polyglycerin # 500, polyglycerin # 750 and the like can be used.

  The polyglycerol having an average degree of polymerization of 2 to 20 calculated from the hydroxyl value of the component (A) is preferably incorporated in an amount of 0.1 to 5.0% by weight in the liposome preparation of the present invention. It is more preferable that it is blended in an amount of 3 to 4.5% by weight. When the above-mentioned polyglycerin is blended in the above ratio, stable liposome can be formed without inhibiting the formation of a single layer or multi layer liposome, and the above active ingredient It can be permeated and stored up to the dermis without being confined to the epidermis of the skin.

  The at least one phospholipid component which is the component (B) of the present invention is a substrate for forming unilamellar or multilamellar liposomes. As the above-mentioned phospholipid component, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl serine, phosphatidyl glycerol, diphosphatidyl glycerol, sphingomyelin etc. are preferable, and one or more of them can be used.

  And, in the liposome preparation of the present invention, it is preferable that 40 to 90% by weight of the at least one kind of phospholipid component which is the component (B) is blended, and further that 50 to 85% by weight is blended. More preferable. By blending the above-mentioned phospholipids in the above proportions, stable unilamellar or multilamellar liposomes can be formed.

  Component (C) of the present invention The active ingredient in at least one pharmaceutical or cosmetic product is a useful compound that brings about healing or improvement on human health, or a preparation containing the compound. And as said active ingredient, a vitamin and its derivative (s), an anti-inflammatory agent, an antiviral agent, an antibacterial agent, an antifungal agent, an analgesic, a spasm, an antioxidant, a moisturizer, a whitening agent, a hair restorer, an anti-wrinkle agent And cell activators are preferred. In addition, one or two or more of them can be used.

  Specifically, as vitamins, fat-soluble vitamins such as vitamin A, vitamin D, vitamin E and vitamin K, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B8, vitamin B9 And vitamin B12 such as vitamin B12 and water-soluble vitamins such as vitamin C, and the like, and their derivatives are also preferable, and one or more of them can be used.

  And, as anti-inflammatory agents, clobetasol propionate, diflorazone acetate, mometasone furan carboxylate, betamezone butyrate ester propionate, fluocinonide, betamethasone dipropionate, diflupredonato, amcinonide, diflucortrone valeric acid Esters, hydrocortisone butyrate, deprodone propionate, dexamethasone propionate, dexamethasone valerate, betamethasone valerate, beclomethasone propionate, fluocinoronacetonide, prednisolone valerate acetate, triamcinolone acetonide, flumeta Zonpivalate, alclomethasone propionate, clobetasone butyrate , Corticosteroids such as hydrocortisone butyrate, dexamethasone, prednisolone, diclofenac sodium, indometacin, indomethacin, ketoprofen, piroxicam, felubinac, loxoprofen sodium hydrate, suprofen, ufenamate, ibuprofen piconol, dimethyl isopropyl azulene, glytyryl retinate Etc., aminobenzoic acid ester, camphor, zinc oxide, calcium hydroxide, crotamiton etc. are preferable, and one or more of them can be used.

  And, as the antibacterial agent, antiviral agents such as vidarabine and acyclovir, chloramphenicol, tetracycline hydrochloride, fradiomycin sulfate, sodium fusidate, gentamycin sulfate, clindamycin ester, nadifloxacin etc are preferable. One or two or more of them can be used.

  And, as an antifungal agent, tolnaphthalate, liranaphthalate, clotrimazole, miconazole nitrate, econazole nitrate, isoconazole nitrate, isoconazole nitrate, sulconazole nitrate, oxonazole nitrate, bifonazole, neticonazole nitrate, ketoconazole, lanaconazole, luliconazole, allomorphin nitrate, terbinafine nitrate Butenafine nitrate, ciclopirox olamine and the like are preferable, and one or more of them can be used.

  And, as the analgesic, glycol salicylate, methyl salicylate, aminobenzoic acid ester, camphor and the like are preferable, and one or more of them can be used.

  And, as the antipruritic agent, aminobenzoic acid ester and crotamiton are preferable, and one or more of them can be used.

  And, as an antioxidant, glutathione, N-acetylcysteine, ascorbic acid, α-tocopherol, butylhydroxyanisole, catechin, quercetin, uric acid, bilirubin, glucose, flavonoids, carotenoids, zeluroblastin, albumin, ferritin, metallotonein, superoxide Dismutase, glutathione peroxidase, catalase, thioredoxin and the like are preferable, and one or more of them can be used.

  And, as the moisturizer, moisturizers such as collagen, hyaluronic acid, ceramide, urea, sodium chondroitin sulfate, amino acids and salts thereof are preferable, and one or more of them can be used.

  And as a skin-whitening agent, hydroquinone, ascorbic acid 2-glucoside, ascorbic acid, tranexamic acid, gentisic acid, arbutin, placenta extract, ellagic acid, kojic acid, ferulic acid, rucinol etc. are preferable, and one or two or more of them. Can be used.

  And, as the hair-growing agent, minoxidil, finasteride, cepharanthine, seembly extract, carpronium chloride and the like are preferable, and one or more of them can be used.

  And, as the anti-wrinkle agent, retinol, retinoic acid and the like are preferable, and one or more of them can be used.

  And as a cell activator, DNA, a placenta extract, a chlorella extract etc. are preferable, and 1 type, or 2 or more types of those can be used.

  And, in the liposome preparation of the present invention, the active ingredient which is the component (C) is preferably incorporated in an amount of 9.9 to 55.0% by weight, and further, 14.7 to 45.5% by weight Is more preferable. When the above active ingredients are blended in the above proportion, stable liposomes can be formed without inhibiting the formation of unilamellar or multilamellar liposomes, and the formation of desired liposomes is also possible. While exerting an effect, it is possible to express a stable effect that does not cause any adverse effects.

  Generally, as a method for producing a liposome, a thin film method, an ultrasonic method, a reverse phase evaporation method, a French press method, a homogenization method, an extrusion method, an ethanol injection method, a dehydration-rehydration method, etc. are used. However, two or more of these methods can be used in combination. In the present invention, a step of dissolving at least one phospholipid component in an organic solvent such as alcohol in a container such as a flask, a step of almost completely volatilizing and removing the organic solvent by heating under reduced pressure, etc. A buffer solution was added to the thin film, stirred, and further subjected to ultrasonic wave irradiation, and the obtained suspension was subjected to pressure passing through micropores and adjusted. According to such a method for producing a liposome, the liposome produced has a phospholipid component formed as a vesicle or the like having a monolayer or bilayer, and the surface is rendered hydrophilic.

  In addition, on the surface of the skin, sebum and sweat discharged from the sweat glands are mixed to form a sebum film to prevent external stimulation, and the sebum film of healthy skin is maintained at a weak acidity of pH 4.5 to 6.0. Therefore, in order to stably contact the prepared liposome preparation on the surface of the skin and permeate the above-mentioned active ingredient contained in the liposome preparation, a charge inducing agent is added when the liposome preparation is prepared. be able to. As the charge inducer, an anionic surfactant having a carboxylic acid, sulfonic acid or phosphoric acid structure as a hydrophilic group is preferable. Specifically, sodium cholate, sodium octanoate, sodium decanoate, sodium laurate, sodium myristate, sodium palmitate, sodium stearate, sodium N-lauroyl sarcosine, sodium cocoyl glutamate, alpha sulfo fatty acid methyl ester salt, etc. Carboxylic acid type, sodium 1-octane sulfonate, sodium 1-decane sulfonate, sodium alkylbenzene sulfonate, sodium naphthalene sulfonate, etc. sulfonic acid type, sodium lauryl sulfate, sodium myristyl sulfate, sodium laureth sulfate, polyoxyethylene alkylphenol Sodium sulfonate, sulfate type such as ammonium lauryl sulfate, lauryl phosphate, sodium lauryl phosphate, And phosphoric acid ester types, such as potassium Urirurin acid.

  Furthermore, in addition to the above components, a preservative such as methyl paraben, a cooling agent such as l-menthol, a fragrance, a pigment, and the like can be added to the liposome preparation of the present invention as required.

Example 1
17.2 mg of phosphatidylcholine (Soya phosphatidylcholine) derived from soybean as a substrate for forming liposome, prepared in advance, 0.59 mg of sodium cholate as a charge inducer, and vitamin C derivative as an active ingredient A mixture of ethanol solutions, each of which has been adjusted in advance to an amount of 2.18 mg of cetyl ascorbate (VC-cetyl ether) and 0.72 mg of tocopherol acetate (Tocopherol acetate) which is a vitamin E derivative (Soya PC) / Sodium cholate / VC-cetyl ether / Tocopherol acetate molar ratio is 80/5/20/6), then the mixture is heated in a water bath at 40 ° C. under reduced pressure using a rotary evaporator to distill off ethanol I left it. Then, 0.1% disodium edetate (EDTA-2Na), 0.4 mg of 50 mM citrate buffer (pH = 5.0), and 0.12 mg of diglycerin in the buffer were added to the remaining thin film. They were added and stirred, and ultrasonication was performed for 5 minutes in a bath-type sonicator to suspend the thin film. Then, an operation of extruding the suspended liquid from a polycarbonate filter with a pore size of 100 nm was performed 10 times using an extruder to prepare a liposome preparation.

Comparative Example 1
A liposome preparation was prepared in the same manner as in Example 1 except that 0.12 mg of glycerol was added instead of 0.12 mg of diglycerin.

Comparative Example 2
A liposome preparation was prepared in the same manner as in Example 1 except that no additive was added instead of 0.12 mg of diglycerin.

  Table 1 shows a list of the compounding amounts of the respective components in the preparation of the liposome preparation of Example 1 and Comparative Examples 1 and 2 and Table 2 shows a list converted in terms of the mixing ratio of each component to the liposome preparation.

<Permeability test and storage ability test>
Using the liposome preparations of Example 1 and Comparative Examples 1 and 2, the amounts of the active ingredients of cetyl ascorbate ascorbic acid which is a vitamin C derivative and tocopherol acetate which is a vitamin E derivative are stored in the epidermis and dermis of the skin. It was measured. The skin is widely used in dermatological research, and was performed using Yucatan Micro Pig skin (hereinafter referred to as YMP skin) close to human skin tissue structure.

Specifically, first, YMP skin which had been stored frozen at -80 ° C in advance was naturally thawed at room temperature, subcutaneous fat was removed with scissors in a half thawed state, and it was further adhered to the dermis with a scalpel for surgery Removed fat. Then, the YMP skin thus treated was attached to a Franz diffusion cell (effective diffusion area: 0.38 cm ² ) such that the stratum corneum side was the donor side and the dermis side was the receiver side. Then, 500 μL of each liposome preparation was placed in the donor compartment, and 5.0 mL of a 30% by weight ethanol solution (pH 7.4) of phosphate buffered saline (PBS) was used as the receiver solution on the reservoir side. During the test, the receiver solution was continuously stirred by a stirrer, and the temperature was maintained at 37 ° C. by means of a thermostatic water pump, and parafilm was used during the test, under the blocking condition with the top of the donor compartment closed. In the test, 200 μl solution is collected from the receiver phase every 0, 1, 2, 4, 6, 24 hours after the start of the test, and cetyl ascorbate contained in each sample (VC-cetyl ether), tocopherol acetate The analysis was carried out by quantifying the amount of ester (Tocopherol acetate) by high performance liquid chromatography. In the retention test, the YMP skin after 24 hours is taken out, the donor solution on the YMP skin is sufficiently wiped off, then separated into the epidermis and the dermis, each is ground using a homogenizer, and centrifuged at 10000 rpm for 10 minutes The supernatant is collected and filtered through a filter, and the filtrate is again treated with the amount of cetyl ascorbate (VC-cetyl ether) and tocopherol acetate (Tocopherol acetate) contained in each sample. It quantified by high performance liquid chromatography.

  High-performance liquid chromatography can be performed using a pump (LC-20AD, Shimadzu Corporation), an auto injector (SIL-20AC, Shimadzu Corporation), a UV detector (SPD-20A, Shimadzu Corporation), and an analysis system (CBM-20A, Shimadzu Corporation) The column was TSK-gel ODS-100Z 3 μm (100 × 2.0 mm, Tosoh Corporation), and the guard column was TSK-gel guardgel ODS-100Z 3 μm (10 × 2.0 mm, Tosoh Corporation). As the mobile phase, methanol (containing 0.1% trifluoroacetic acid): water (containing 0.1% trifluoroacetic acid) was used at a ratio of 97: 3 (v / v). The flow rate was 0.2 mL / min, the UV wavelength was 246 nm (for detecting cetyl ascorbate), 285 nm (for detecting tocopherol acetate), and the column temperature was 40 ° C. In this condition, the retention time of cetyl ascorbate (VC-cetyl ether) was 2.5 minutes, and that of tocopherol acetate was 10 minutes. For quantification, cetyl ascorbate (VC-cetyl ether) and tocopherol acetate (Tocopherol acetate) were quantified using ergocalciferol as an internal standard and using an internal standard method.

  As a result, in the permeability test, each sample collected from the receiver phase every 0, 1, 2, 4, 6 or 24 hours after the start of the test was cetyl ascorbate (VC-cetyl ether), tocopherol acetate Since the ester (Tocopherol acetate) is below the detection limit, it can be judged that neither ascorbyl cetyl ether (VC-cetyl ether) nor tocopherol acetate (Tocopherol acetate) permeated the YMP skin.

  And, in the retention test, actual accumulation of cetyl ascorbate ascorbic acid (VC-cetyl ether) or tocopherol acetate (Tocopherol acetate) in the epidermis or dermis when the liposome preparations of Example 1 and Comparative Examples 1 and 2 are used. The amount was determined by performing each test six times, and from the results, the arithmetic mean was determined after converting to the accumulated amount (ng) per unit weight (mg) of the epidermis or dermis. The results for each of the accumulated amounts (ng) of cetyl ascorbate (VC-cetyl ether) or tocopherol acetate (Tocopherol acetate) per unit weight (mg) of the epidermis or dermis are shown in Table 3 and Table 4.

  As shown in Table 3 and Table 4 and FIG. 1 and FIG. 1, cetyl ascorbate (VC-cetyl ether) and tocopherol acetate (Tocopherol acetate) penetrate the epidermis and reach the dermis and are stored. As compared with Comparative Example 1 in which glycerin was blended, and Comparative Example 2 in which nothing was blended, it was found that the largest amount was stored in the dermis in Example 1 in which diglycerin was blended.

Claims (3)

(A) polyglycerin having an average degree of polymerization of 2 to 20, which is calculated from hydroxyl value,
(B) at least one phospholipid component,
(C) A pharmaceutical or cosmetic liposome preparation comprising an active ingredient in at least one pharmaceutical or cosmetic product. The liposome preparation according to claim 1, wherein the phospholipid component is at least one selected from phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl serine, phosphatidyl glycerol, diphosphatidyl glycerol and sphingomyelin. The said active ingredient is a vitamin and its derivative, an anti-inflammatory agent, an antiviral agent, an antibacterial agent, an antifungal agent, an analgesic agent, an analgesic agent, an antipruritic agent, an antioxidant, a moisturizer, a whitening agent, a hair restorer, an anti-wrinkle agent, cell activation The liposome preparation according to claim 1 or 2, which is at least one selected from agents.

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