JP2011231128A - Skin external preparation for normalizing horny layer cell differentiation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a pharmaceutical composition for external use, that synergistically facilitating normal differentiation of a horny layer cell by facilitating an action of increasing an area of the horny layer cell.SOLUTION: There is provided the pharmaceutical composition for external use, that includes: heparinoid; allantoin or a derivative thereof; and pantothenic acid or the derivative thereof.

Description

  The present invention relates to an external pharmaceutical composition that promotes normal differentiation of epidermal keratinocytes into stratum corneum cells.

  The stratum corneum of the skin exists in the outermost layer of the living body, and has a barrier function such as protection against excessive water evaporation from the living body and contact between the stimulating substance or chemical substance and the living body. The stratum corneum cells that make up the stratum corneum migrate to the superficial layer after the epidermal keratinocytes proliferate in the basal layer, differentiate into spinous layers and granule layers, and morphologically enucleate and flatten during final differentiation Formed through change. The form is a flat plate-like shape. For example, when differentiation of epidermal keratinocytes is not normally performed, abnormally shaped horny layer cells are produced. The area is generally known to be inversely proportional to the differentiation rate of epidermal keratinocytes. The differentiation rate of epidermal keratinocytes is enhanced by disruption of the barrier function or stimulation such as inflammation, resulting in immature cells with small areas and thick shapes. It is known that the stratum corneum composed of such abnormal stratum corneum cells is inferior to the normal stratum corneum in the barrier function that it should originally have. Therefore, the measurement of the stratum corneum cell area is useful for distinguishing the normality of the stratum corneum cells and the differentiation state of the epidermal keratinocytes, and the skin state can be estimated from the results (Patent Documents 1 and 2). That is, normalizing the area of stratum corneum cells means normalization of epidermal keratinocyte differentiation and normalization of the epidermal barrier function associated therewith.

  Conventionally, when differentiation is abnormal in epidermal keratinocytes, a method of using a moisturizing agent such as an external preparation for skin as a symptomatic treatment for the resulting dry skin has been taken. Commonly used moisturizers include polyhydric alcohols such as glycerin, 1,3-butylene glycol, sorbitol, natural plant-derived extract components, amino acids that are skin constituents, natural moisturizing factors such as pyrrolidone carboxylic acid, ceramides, Examples include intercellular lipid components such as cholesterol, and in vivo polymers such as collagen and hyaluronic acid. By using a topical skin preparation containing these, improvement of skin moisture is attempted by improving the skin's moisture retention. It was. However, the application of such an external preparation has only an effect of temporarily increasing the moisture content of the skin, and it takes a long time to recover, so that essential recovery of the skin function cannot be expected. An external preparation for skin having a normalizing action has been desired.

  On the other hand, there is a heparin-like substance as a substance having a moisturizing function blended in a skin external preparation. Heparin-like substances are known to have a blood circulation promoting action and a fibroblast proliferation inhibiting action. In addition, heparin-like substances are used as therapeutic agents for various dry skin diseases such as senile xeroderma that have a moisturizing ability, but reports that they do not affect stratum corneum cells and barrier function. (Non-Patent Document 1).

JP 2003-38449 A JP 2001-13138

Journal of Japanese Society of Clinical Dermatology, 56 ： 87-96, 1998

  The present invention provides an external pharmaceutical composition that can promote normal differentiation into stratum corneum cells, increase the area of the stratum corneum cells constituting the stratum corneum, and normalize the stratum corneum structure and function.

As a result of diligent research, the inventor has promoted normal differentiation into horny layer cells to increase the area of horny cells constituting the horny layer, thereby maintaining or normalizing the structure and function of the horny layer. The present invention has been completed.
That is, this invention is an external pharmaceutical composition characterized by including an allantoin and / or allantoin derivative, and panthenol and / or a panthenol analog in a heparin analog.
In particular, in the pharmaceutical composition for external use of the present invention, the content of heparin-like substance is 0.01-5% by mass of the whole, the content of allantoin and / or allantoin-related substance is 0.01-5% by mass of the whole, panthenol and / or panthenol. The content of related substances is 0.01 to 10% by mass of the whole.

INDUSTRIAL APPLICABILITY According to the present invention, an external pharmaceutical composition suitable for skin diseases such as atopic dermatitis associated with abnormal differentiation of epidermal keratinocytes and mild rough skin can be provided.
According to the present invention, the shape and cell area of the horny layer cells that are abnormal due to skin diseases and the like are sufficiently close to the shape and area that the horny layer cells should originally have, and the structure and function that the horny layer should originally have ( For example, the effect of the present invention is “the effect of promoting normalization of stratum corneum cell differentiation”, “the effect of promoting normalization of the stratum corneum cell area” or “the effect of promoting normalization of the stratum corneum”. It can also be called. Therefore, the pharmaceutical composition for external use of the present invention can also be referred to as “keratinocyte differentiation normalizing agent”.
The external pharmaceutical composition of the present invention has an enhanced effect of improving or treating the condition of the affected area.

FIG. 1 shows the stratum corneum cell area in pixels when the composition of the present invention and the composition of the comparative example are applied (Example 1). * p <0.05 (n = 8) FIG. 2 shows the stratum corneum cell area in pixels when the composition of the present invention and the composition of the comparative example are applied (Example 2). ** P <0.01, * P <0.05 (n = 8)

  The present invention synergizes normal differentiation of stratum corneum cells when a heparin-like substance used as a moisturizing agent is combined with allantoin and / or allantoin-related substances, panthenol and / or panthenol-related substances, which are wound healing promoters. Based on finding out to promote. The heparin-like substance used in the present invention is a polysulfated ester of mucopolysaccharide, which is a polysulfated polysaccharide having a repeating unit of a disaccharide composed of D-glucuronic acid and N-acetyl-D-galactosamine. . Those having many hydrophilic groups such as sulfate group, carboxyl group and hydroxyl group in the structure, having high moisturizing ability and listed in the Japanese Pharmacopoeia Standards for Drugs are preferably used.

  In the present invention, allantoin and allantoin-related substances are active ingredients having an effect of promoting wound healing. Allantoin-related substances are active ingredients that have a hydantoin skeleton and have wound healing and anti-inflammatory effects. Specifically, allantoin-related substances include, but are not limited to, allantoin chlorohydroxyaluminum and allantoindihydroxyaluminum. .

  Panthenol and panthenol-related substances used in the present invention are active ingredients having a wound healing action. Specifically, panthenol-related substances include pantothenyl ethyl ether, calcium pantothenate, sodium pantothenate, acetyl Including but not limited to panthenyl ethyl ether.

In the external pharmaceutical composition of the present invention, the above-mentioned heparin-like substance can be 0.01 to 5% by mass, preferably 0.05 to 3% by mass based on the total. Allantoin and / or allantoin-related substances can be 0.01 to 5% by mass, preferably 0.1 to 3% by mass, based on the total amount. Panthenol and / or panthenol-related substances may be 0.01 to 10% by mass, preferably 0.1 to 5% by mass, based on the total.
In the pharmaceutical composition for external use of the present invention, heparin-like substances, allantoin and / or allantoin derivatives, panthenol and / or panthenol analogues can be blended in two or more, respectively, in which case the total amount for each Is preferably blended in such a range as described above.

  Evaluation of the effects of the present invention (“stratification of stratum corneum cell differentiation normalization”, “promotion of stratum corneum cell area normalization” or “promotion of stratum corneum normalization”) was carried out using an appropriate experimental model known to those skilled in the art. Can be used. Specifically, for example, evaluation can be performed as described in the following 1) to 6).

1) Put the test site on the back of the neck of hairless mouse with absorbent cotton impregnated with acetone / ether (1: 1) and purified water, destroy the skin barrier, and create an animal model with a small stratum corneum cell area.
2) Apply 20 mg of the sample such as the external pharmaceutical composition to the test site of the animal model.
3) The stratum corneum cells are collected from the test site by the tape stripping method.
4) The collected horny layer cells are stained with brilliant green / gentian violet (BG) staining.
5) Microscopic images of stained stratum corneum cells are analyzed by image processing software (for example, ImageJ: developed by National Institutes of Health), and the area of stratum corneum cells is measured. At that time, a fixed number of stratum corneum cells for each sample, for example, 30 stratum corneum cells are randomly selected, the area is averaged, and the standard deviation is calculated in some cases, and these values are used as the stratum corneum cell area in each sample. Can be evaluated.
6) The effect of each specimen can be evaluated by comparing the stratum corneum cell area for each specimen with the stratum corneum cell area of the uncoated control.
In the present specification, “normal differentiation of stratum corneum cells” means that epidermal keratinocytes that have proliferated in the basal layer become flat stratum corneum cells by differentiation into spinous layers and granular layers. It is a process of moving to the skin surface, and is a process in which cells having a morphology and a cell area that cells normally constituting the stratum corneum (corneal cells) appear in the epidermis in an untreated healthy control animal.

  The external pharmaceutical composition of the present invention includes heparin-like substances, allantoin and / or allantoin derivatives, panthenol and / or panthenol-related substances, as well as pharmaceuticals and quasi drugs as long as the effects of the invention are not impaired as required. Various active ingredients used for cosmetics and the like may be blended. For example, it can contain anti-inflammatory agents, antihistamines, herbal medicines, antipruritic agents, wound healing agents, local anesthetics, vitamins, cooling agents, moisturizers, bactericides, vasoconstrictors, amino acids, etc. The following components are exemplified.

Examples of anti-inflammatory agents include glycyrrhetinic acid, glycyrrhizic acid or derivatives thereof, licorice extract, steroid compounds (hydrocortisone, prednisolone, methylprednisolone, clobetasone, betamethasone, dexamethasone, cortisone, flumethasone, beclomethasone, fluticasone or derivatives thereof), indomethacin , Ibuprofen, ibuprofen piconol, bufexamac, ufenamate, piroxicam, ketoprofen, salicylic acid or a derivative thereof, dimethylisopropylazulene, toki extract, sicon extract and the like.
Examples of the antihistamine include diphenhydramine, chlorpheniramine, mequitazine, azelastine, emedastine, ketotifen, and derivatives thereof. Preferred are diphenhydramine hydrochloride and chlorpheniramine maleate.

  Examples of the whitening agent include L-cysteine, hydroquinone, glucosamine, L-ascorbic acid, glutathione, kojic acid, ellagic acid, placental extract, ubiquinones or derivatives thereof.

Examples of herbal ingredients are Psycho, Bukuryo, Keihi, Kanzo, Ogon, Oubak, Ouren, Sanshishi, Giou, Peonies, Senkyuu, Toki, Hamaboufu, Bofuu, Ohi, Kyoko, Gankyo, Dokkatsu, Keigai, Mokutsumo, Ushiboshi, , Centai, Kujin, Sojitsu, Inchinkou etc.
Examples of antipruritic agents include salicylic acid or its derivatives, nonylic acid vanillylamide, capsaicin, benzyl nicotinate, chili tincture and the like.
Examples of wound healing agents include zinc oxide.
Examples of local anesthetics include lidocaine, dibucaine, procaine, tetracaine, aminobenzoic acid, or derivatives thereof.

  Examples of vitamins include vitamins A [retinol and its derivatives (eg, retinal, retinoic acid, retinol palmitate, etc.)], vitamin B1 [thiamine and its derivatives (eg, thiamine hydrochloride, thiamine nitrate, etc.)], Vitamin B2s [riboflavin and its derivatives (for example, riboflavin phosphate, sodium riboflavin phosphate, riboflavin butyrate, and flavin adenine dinucleotide sodium)], vitamin B3 [nicotinic acid and its derivatives (nicotinic acid amide, tocopherol nicotinate) , Benzyl nicotinate, etc.)], vitamin B6s [pyridoxine and its derivatives (eg, pyridoxine hydrochloride, pyridoxine phosphate, pyridoxal, etc.)], vitamin B12 [cobalamin and its derivatives (eg, shea Cobalamin, mecobalamin, and hydroxocobalamin hydrochloride, etc.)], biotin, folic acid or a pharmaceutically acceptable salt thereof, vitamin C [ascorbic acid and its derivatives (for example, erythorbic acid, sodium ascorbate, alcorbic acid palmitate, etc.), Vitamin Ds [calciferol and its derivatives (eg, ergocalciferol, cholecalciferol, etc.)], vitamin E [tocopherol, ubiquinone and its derivatives (eg, tocopherol acetate, tocopherol calcium succinate, etc.)], other vitamins (For example, hesperidin, carnitine, ferulic acid, γ-oryzanol, orotic acid, rutin, eriocitrin, inositol, and pharmaceutically acceptable salts thereof).

Examples of the refreshing agent include camphor, borneol or related substances thereof, fennel oil, eucalyptus oil, peppermint oil, and the like.
Examples of humectants include polyhydric alcohols (such as glycerin and 1,3-butylene glycol), hyaluronic acid or derivatives thereof, polymer compounds (such as collagen and chitosan), amino acids (such as glycine, alanine, and aspartic acid), and natural moisturizing agents. Factors (sodium lactate, urea, etc.), ceramide, plant extract (chamomile extract, aloe extract, etc.) and the like can be mentioned.

Examples of fungicides include isopropylmethylphenol, chlorhexidine hydrochloride, benzethonium chloride, benzalkonium chloride, cetrimide and the like.
Examples of the vasoconstrictor include naphazoline, tetrahydrozoline, methylephedrine or salts thereof.
Examples of amino acids include glutamic acid, aspartic acid, glycine, alanine, serine, aminoethylsulfonic acid (taurine), and pharmaceutically acceptable salts thereof (for example, ephedrine hydrochloride, methylephenoline hydrochloride, etc.) and the like. .

The pharmaceutical composition for external use of the present invention can be prepared by blending the above-mentioned heparin-like substance, allantoin and / or allantoin derivative, panthenol and / or panthenol-related substance and various bases according to a conventional method. . Any of the raw materials generally used as a base can be used for preparing the external pharmaceutical composition of the present invention.
Examples of such bases include hydrocarbons such as petrolatum, paraffin, liquid paraffin, squalane, ceresin, gelled hydrocarbon and microcrystalline wax; stearic acid, isostearic acid, myristic acid, oleic acid, etc. and behenic acid, etc. Higher fatty acids such as cetanol, stearyl alcohol and behenyl alcohol; fatty acid ester oils such as isopropyl myristate, octyldodecyl myristate, isopropyl palmitate and diisopropyl adipate; glycerin triisooctanoate and tri (capryl-capric acid) Polyhydric alcohol fatty acid esters such as glyceryl; glyceryl monostearate, glyceryl monooleate, sorbitan monostearate, sorbitan monooleate, sesquiolei Sorbitan acid, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (60) sorbitol tetrastearate, polyoxyethylene (60) sorbit tetraoleate, polyoxyethylene (60) castor oil, polyoxyethylene (60) hydrogenated castor oil, polystearic acid monooxyethylene glycol (4EO), polyethylene glycol monooleate (10EO), polyoxyethylene (9) lauryl ether, polyoxyethylene (10 ) Nonionic surfactants such as cetyl ether and polyoxyethylene oleyl ether; ionic surfactants such as sodium lauryl sulfate and sodium cetyl sulfate; propylene glycol, 1,3-butylene glycol and polyethylene glycol Polyhydric alcohols such as carboxy vinyl polymer, hydroxypropyl cellulose, hypromellose, hydrophobized hydroxypropyl methyl cellulose, acrylated starch 300, and polyvinyl pyrrolidone; diisopropanolamine, triethanolamine, sodium hydroxide, hydrochloric acid, PH adjusting agents such as citric acid, sodium citrate, potassium dihydrogen phosphate and sodium hydrogen phosphate; organic solvents such as ethanol, isopropyl alcohol, acetone, methyl ethyl ketone and ethyl acetate; sodium chloride, sodium thiosulfate, sodium sulfite and edet Stabilizers such as sodium acid; preservatives such as methylparaben, propylparaben, benzalkonium chloride, and benzethonium chloride, but are not limited thereto.

  When preparing the pharmaceutical composition for external use of the present invention, it is prepared by mixing the above ingredients and a medium such as water, but the mixing ratio is not limited, for example, 0 to 80% by mass of water, 0 to 0 to hydrocarbons 99 mass%, higher fatty acid 0-25 mass%, higher fatty alcohol 0-25 mass%, fatty acid ester oil 0-50 mass%, polyhydric alcohol fatty acid ester 0-30 mass%, surfactant 0-10 mass%, Polyhydric alcohol 0-90 mass%, polymer compound 0-10 mass%, pH adjuster 0-10 mass%, organic solvent 0-99 mass%, stabilizer 0-10 mass%, preservative 0-5 mass %.

  The pharmaceutical composition for external use of the present invention can be produced according to a method for producing ordinary ointments, creams, emulsions, gels, liquids for external use, lacquers, patches, poultices, suppositories, and injections. The preparation method is not particularly limited. For example, each preparation is prepared by blending appropriate ingredients such as known excipients necessary for preparing preparations of various dosage forms together with the above essential ingredients and optional ingredients. It can be prepared by conventional methods.

  The amount and usage of the external pharmaceutical composition of the present invention are not particularly limited, and can be appropriately selected depending on the dosage form and the like, for example, for keratinization failure associated with dry skin disease and inflammatory skin disease. Dry skin for 1 to several days, usually 1 to several times a day with the usual amount of various external preparations for skin (external pharmaceutical composition) used for treatment or care, such as conventional pharmaceuticals, quasi-drugs, and cosmetics Apply to skin where keratosis failure or keratosis dysfunction is caused by the above-mentioned disease by applying uniformly on the affected area where illness or inflammatory skin disease is occurring or in the vicinity. Thus, it is possible to effectively improve or prevent a keratosis condition associated with dry skin diseases and inflammatory skin diseases represented by atopic dermatitis and housewife eczema.

  EXAMPLES Hereinafter, although an Example and a comparative example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to the following Example.

The stratum corneum area normalizing action of an external solution containing allantoin and panthenol in a heparin-like substance and an external solution containing a heparin-like substance alone and an external solution containing allantoin and panthenol were compared.
[Preparation of Composition of Example and Composition of Comparative Example]
The following compositions were prepared. The composition of each composition is shown in Table 1. In the table, the numerical value regarding each component is the mass% of each component with respect to the whole composition.
1) Composition of Example 1 Add heparin analogs, allantoin and panthenol as active ingredients to purified water and dissolve to make uniform. Next, a solution prepared by adding citric acid and sodium citrate to purified water was added to obtain a solution for external use. The blending amounts and base components are shown in Table 1.
2) Composition of comparative example 1 Except not adding a heparin analog, operation similar to preparation of the composition of Example 1 was performed, and the liquid for external use was obtained.
3) Composition of comparative example 2 Except not adding allantoin and panthenol, operation similar to preparation of the composition of Example 1 was performed, and the liquid for external use was obtained.

Table 1

[Measurement of stratum corneum normalization promoting effect by mouse of each composition]
Eight male hairless mice (weight approx. 30 g, 7 weeks old) per group in a room adjusted to 21.0 ± 0.5 ° C and humidity 55 ± 5% on the back skin with an equal volume of acetone and ether Distilled water was applied twice a day for 5 days, and the dorsal skin of hairless mice with sufficiently small stratum corneum cells was used as the test site. 20 μL of the topical solutions of Examples and Comparative Examples were applied to the test site at the test site once a day for a total of 3 times on each back. Moreover, the purified water was applied by the same operation as object.
The day after application, stratum corneum cells are collected from the test site by tape stripping and stained with brilliant green / gentian violet (BG), and then the microscopic images of the stained stratum corneum cells are image processing software (ImageJ : Developed by National Institutes of Health), the observation images of stratum corneum cells were analyzed, about 30 stratum corneum cells were randomly selected for each sample, the area was averaged, and the corneum cell area in each sample was evaluated.

＊p < 0.05 vs対照
表２及び図１から明らかなように、実施例１の群では各比較例群に対して角層を構成する角層細胞面積が有意に大きくなった。角層細胞の正常な分化が促進され、角層正常化が特に促進されていることが示され、ヘパリン類似物質とアラントイン・パンテノールが組み合わさることで、ヘパリン類似物質に対するアラントイン及びパンテノール配合による相乗効果が明らかになった。 (Test results)
The results are shown in Table 2 and FIG.
Table 2 Effect of promoting normalization of stratum corneum by mice (n = 8)

* P <0.05 vs control As is clear from Table 2 and FIG. 1, the stratum corneum cell area constituting the stratum corneum was significantly increased in the group of Example 1 with respect to each comparative example group. It is shown that normal differentiation of stratum corneum cells is promoted, and normalization of stratum corneum is particularly promoted. By combining heparin analogs and allantoin panthenol, allantoin and panthenol are added to heparin analogs. A synergistic effect was revealed.

  External cream containing allantoin and panthenol in heparin analog, external cream containing heparin analog alone, external cream containing allantoin and panthenol, heparin analog, allanin and panthenol free The effect of normalizing the stratum corneum area was compared.

[Preparation of Composition of Example and Composition of Comparative Example]
The following compositions were prepared. The composition of each composition is shown in Table 3. In the table, the numerical value regarding each component is the mass% of each component with respect to the whole composition.
1) Composition of Example 2 As a preparation method, first, tocopherol acetate, stearyl alcohol, behenic acid, cetyl palmitate, microcrystalline wax, glycerin triisooctanoate, white petrolatum, sorbitan monostearate, glycerin fatty acid ester and poly Oxyethylene cetyl ether is heated and dissolved uniformly. Next, add purified water to heparin-like substance, panthenol, allantoin, glycerin, 1,3-butylene glycol, sodium edetate, citric acid and sodium citrate, heat to dissolve uniformly, and stir into the previous oil phase. While added, an emulsion was prepared. This was further cooled to 40 ° C. or lower to obtain an external cream. The blending amounts and other base components are shown in Table 3.
2) Composition of Comparative Example 3 An external cream was obtained in the same manner as in the preparation of the composition of Example 1 except that no heparin-like substance was added.
3) Composition of comparative example 4 Except not adding allantoin and panthenol, operation similar to preparation of the composition of Example 1 was performed, and the cream for external use was obtained.
4) Composition of Comparative Example 5 A cream for external use was obtained by performing the same operation as in the preparation of the composition of Example 1 except that the heparin-like substance, allantoin and panthenol were not added.

Table 3

  The effect of promoting the normalization of the stratum corneum by mice of each composition was performed in the same manner as described in Example 1. The control was purified water as in the example.

＊＊p < 0.01、＊p < 0.05 vs比較例
表４及び図２から明らかなように、実施例２の群では各比較例群に対して有意に大きくなった。角層細胞の正常な分化が促進され、角層正常化が特に促進されていることが示され、ヘパリン類似物質とアラントイン・パンテノールが組み合わさることで、ヘパリン類似物質に対するアラントイン及びパンテノール配合による相乗効果が明らかになった。 (Test results)
The results are shown in Table 4 and FIG.
Table 4 Effect of promoting normalization of stratum corneum by mice (n = 8)

** p <0.01, * p <0.05 vs Comparative Example As is clear from Table 4 and FIG. 2, the group of Example 2 was significantly larger than each comparative group. It is shown that normal differentiation of stratum corneum cells is promoted, and normalization of stratum corneum is particularly promoted. By combining heparin analogs and allantoin panthenol, allantoin and panthenol are added to heparin analogs. A synergistic effect was revealed.

Claims (4)

  An external pharmaceutical composition comprising a heparin-like substance, allantoin and / or allantoin derivative, and panthenol and / or panthenol-related substance.   The external pharmaceutical composition according to claim 1, wherein the allantoin-related substance is selected from the group consisting of allantoinchlorohydroxyaluminum and allantoindihydroxyaluminum.   The external pharmaceutical composition according to claim 1 or 2, wherein the panthenol-related substance is selected from the group consisting of pantothenyl ethyl ether, calcium pantothenate, sodium pantothenate, and acetyl pantothenyl ethyl ether.   The external pharmaceutical composition according to any one of claims 1 to 3, which is a normalizing agent for horny cell differentiation that promotes normal differentiation of horny layer cells.

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