External composition

Abstract

To provide an external composition used in the inhibition, protection, prevention, or improvement of skin diseases.SOLUTION: An external composition used in the inhibition, protection, prevention, or improvement of skin diseases contains (A) heparinoid, (B) one or more kind selected from the group consisting of allantoin, pantothenic acid, vitamin A and the like, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable derivative thereof, and(C) nonionic surfactant.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for external use used for suppressing, preventing, prophylaxis or amelioration of skin diseases. [Background technology]

[0002] Known skin diseases accompanied by itching include dry skin diseases such as senile xerosis, atopic dermatitis, and housewives' eczema, inflammatory skin diseases, skin pruritus, heat rash, chilblains, cracks, and chapped skin. The skin of patients suffering from these skin diseases becomes sensitive to external stimuli, which is one of the causes of repeated itching. In response to this itching, patients often engage in excessive scratching behavior.

[0003] To treat these symptoms, topical skin compositions containing as active ingredients antipruritic ingredients such as antihistamines and crotamiton, moisturizing agents such as heparinoids and urea, and anti-inflammatory ingredients such as allantoin and glycyrrhizinate are widely used.

[0004] Of these, heparinoids have skin moisturizing and blood circulation promoting effects. Specifically, the skin moisturizing effect of heparinoids reduces dry skin symptoms. In addition, the blood circulation promoting effect of heparinoids promotes blood circulation at the application site, reducing pain and swelling due to blood circulation disorders. For this reason, heparinoids are used to treat or prevent asteatosis, progressive keratoderma tylodes palmaris, chilblains, hypertrophic scars and keloids, pain or inflammatory diseases due to blood circulation disorders, thrombophlebitis, swelling, hematoma, tendonitis, muscle pain, arthritis, etc. after trauma (bruises, sprains, contusions).

[0005] Patent Document 1 describes that a skin topical composition containing crotamiton and a heparinoid can improve itching associated with dry skin diseases, inflammatory skin diseases, and the like.

[0006] Patent Document 2 describes that an external skin composition containing an antihistamine-containing preparation and a heparinoid can improve itching associated with dry skin diseases, inflammatory skin diseases, and the like.

[0007] Patent Document 3 describes that an external skin composition containing a nonsteroidal anti-inflammatory agent and a heparinoid can improve itching associated with dry skin diseases, inflammatory skin diseases, and the like.

[0008] Patent Document 4 discloses a skin topical composition containing a heparinoid, allantoin, and pantothenic acid, and states that this skin topical preparation can promote normal differentiation of stratum corneum cells.

[0009] Patent Document 5 discloses a skin topical composition containing vitamin A, edetic acid or a salt thereof, and a heparinoid, and describes that this skin topical composition can stably contain vitamin A.

[0010] However, excessive scratching behavior of patients in response to itching may cause skin damage or inflammation, leaving itching marks on the skin. In some cases, these itching marks are accompanied by pigmentation. Other cases in which pigmentation occurs on the skin include cases where pigmentation occurs (also called scars, etc.) after external wounds due to burns or trauma have healed completely. In addition, hormone imbalance, aging, sudden strong sunburn, chronic friction, etc. can also cause pigmentation on the skin. It is desirable to suppress, prevent, prevent, or improve skin pigmentation from the viewpoint of skin appearance or beauty. It is also known that melanocytes present in the dermis produce excessive melanin, resulting in bruises (bruises) such as black bruises, brown bruises, and blue bruises. Furthermore, when the skin is injured by trauma (bruises, sprains, contusions, etc.), not only pigmentation occurs, but also hematomas (blue bruises due to subcutaneous bleeding) occur, which can cause noticeable pigmentation.

[0011] A hematoma is a condition in which blood pools in the tissue due to the rupture of capillaries in the skin or subcutaneous tissue caused by trauma, and the symptom of a subcutaneous hematoma is a blue or purple discoloration of the injured area. Although hematomas gradually disappear as blood vessels and tissues are repaired and exudates are removed, they may take a long time to recover due to aging and pigmentation. For this reason, from the perspective of skin appearance or cosmetics, it is desirable to quickly eliminate hematomas or quickly fade (lighten) the color of hematomas. For this reason, there has been a demand for suppression, prevention, prophylaxis, or improvement of pigmentation or hematomas. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] JP 2000-229852 A [Patent Document 2] JP 2000-229884 A [Patent Document 3] JP 2000-38352 A [Patent Document 4] JP 2011-231128 A [Patent Document 5] JP 2012-36183 A Summary of the Invention [Problem to be solved by the invention]

[0013] An object of the present invention is to provide a topical composition that can be used to suppress, prevent, prophylactically, or ameliorate skin diseases. [Means for solving the problem]

[0014] The topical composition used to suppress, prevent, prevent or improve skin diseases that solves the above problems contains (A) a heparinoid, (B) one or more substances selected from the group consisting of allantoin, pantothenic acid, vitamin A, pharma- ceutical acceptable salts thereof, and pharma-ceutical acceptable derivatives thereof, and (C) a nonionic surfactant.

[0015] According to the above-mentioned configuration, the composition for external use contains the components (A), (B) and (C), and thus exerts a synergistic effect of inhibiting melanin production, thereby suppressing, preventing, preventing or improving pigmentation of the skin. Effect of the Invention

[0016] According to the present invention, the topical composition contains components (A), (B) and (C), and the synergistic effect of these components can suppress, prevent, prevent or ameliorate pigmentation of the skin. [Brief description of the drawings]

[0017] [Figure 1] 1 is a graph showing the relationship between the amount of polyoxyethylene hydrogenated castor oil 50 blended and the rate of decrease in melanin production. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The preferred embodiments of the present invention will be described in detail below. In this specification, when indicating the relative amount of a component added to a topical composition with respect to a specific component, the unit of "parts by weight" is used.

[0019] The topical composition E of this embodiment is used as a skin medication to be applied to areas affected by dry skin diseases such as senile xerosis, atopic dermatitis, and housewives' eczema, inflammatory skin diseases, skin pruritus, prickly heat, chilblains, cracks, and chapped skin, etc.

[0020] Composition for external use E contains as active ingredients the ingredients (A), (B) and (C) described below.

[0021] The above component (A) is a heparinoid, which is known to have blood circulation promoting effects and fibroblast proliferation inhibiting effects.

[0022] The heparinoid contained in the external composition E is a general term for polysulfated mucopolysaccharides such as chondroitin polysulfate. The heparinoid preferably has an average of 0.5 to 5 molecules, and more preferably an average of 0.6 to 3 molecules, of sulfate groups per monosaccharide molecule constituting the mucopolysaccharide.

[0023] As the heparinoid, a commercially available product that is generally available may be used as it is. Heparinoids can also be obtained by a method of sulfated mucopolysaccharides or a method of extracting naturally occurring sulfated polysaccharides. As a method of extracting naturally occurring sulfated polysaccharides, for example, there is a method of extracting and purifying from the internal organs including the bronchi of animals such as cows and pigs using an aqueous carrier, and then sulfate the extract as necessary. As the heparinoid, those listed in the Japanese Pharmacopoeia Non-Drug Standards are preferably used.

[0024] Furthermore, there is no particular limitation on the molecular weight of the heparinoid, but it is usually about 1,000 to 10,000,000, and preferably 5,000 to 1,000,000.

[0025] The proportion of the amount of component (A) in the composition for external use E to the total amount of the composition for external use E is not particularly limited as long as the effect of the present application can be achieved, but is, for example, 0.00001% by weight or more. The proportion of the amount of component (A) in the total amount of the composition for external use E is preferably 0.001% by weight or more, more preferably 0.01% by weight or more, even more preferably 0.05% by weight or more, even more preferably 0.1% by weight or more, and particularly preferably 0.3% by weight or more. In a specific embodiment, the proportion of the amount of component (A) in the total amount of the composition for external use E may be 0.45% by weight or more. A preferable effect can be obtained by making the proportion of the amount of component (A) in the total amount of the composition for external use E 0.00001% by weight or more. An excellent effect can be obtained by making the proportion of the amount of component (A) in the total amount of the composition for external use E 0.001% by weight or more, and an even more excellent effect can be obtained by making it 0.01% by weight or more. The proportion of component (A) in the total amount of topical composition E is preferably 10% by weight or less, more preferably 7% by weight or less, even more preferably 5% by weight or less, and particularly preferably 1% by weight or less. When the proportion of component (A) in the total amount of topical composition E is less than 10% by weight, the sticky feeling during use of topical composition E is suppressed, and a more excellent feeling of use is obtained.

[0026] Ingredient (B) is one or more selected from the group consisting of allantoin, pantothenic acid, vitamin A, pharma- ceutical acceptable salts thereof, and pharma- ceutical acceptable derivatives thereof. Allantoin is an active ingredient having a wound healing promoting effect. Pantothenic acid has an effect of assisting the metabolism of energy such as carbohydrates, proteins, and lipids, and is known to exert useful physiological activities such as activating cells, activating metabolism, moisturizing, and enhancing resistance. Vitamin A is an active ingredient having an effect of promoting the disappearance of bruises caused by subcutaneous bleeding due to bruises, or fading of bruises. Ingredient (B) may be used alone or in any combination of two or more.

[0027] Specific examples of allantoin salts to be incorporated in the topical composition E include allantoin chlorohydroxyaluminum, allantoin dihydroxyaluminum, etc.

[0028] Specific examples of allantoin derivatives that can be incorporated into the topical composition E include allantoic acid, dihydroxyaluminum allantoinate, allantoin glycyrrhetin, and chlorohydroxyaluminum allantoinate.

[0029] Specific examples of the salt of pantothenic acid to be incorporated in the composition for external use E include calcium pantothenate and sodium pantothenate.

[0030] Specific examples of the pantothenic acid derivatives contained in the topical composition E include panthenol, pantothenyl ethyl ether, acetylpantothenyl ethyl ether, etc.

[0031] The vitamin A compound contained in the composition for external use E is preferably retinol or an ester of retinoic acid, such as retinol acetate, retinol palmitate, or δ-tocopherol retinoate.

[0032] The vitamin A group may be either one isolated from natural products such as animal materials or one chemically synthesized. The vitamin A group may also be used in the form of vitamin A oil. The vitamin A oil may be a natural oil extracted and refined from animals, or may be one in which vitamin A group is dissolved in vegetable oil or the like. A representative example of the latter is vitamin A oil described in the Japanese Pharmacopoeia (containing 30,000 vitamin A units (IU) or more per gram).

[0033] The proportion of the amount of component (B) blended in the composition for external use E (when two or more components are blended as component (B), the sum of the amounts of those blended) to the total amount of the composition for external use E is, for example, 0.0001% by weight or more. The proportion of the amount of component (B) blended in the composition for external use E to the total amount of the composition for external use E is preferably 0.001% by weight or more, more preferably 0.04% by weight or more, even more preferably 0.05% by weight or more, even more preferably 0.5% by weight or more, and particularly preferably 1.5% by weight or more. By making the proportion of the amount of component (B) blended in the composition for external use E to be 0.0001% by weight or more, a preferable effect can be obtained, and by making it 0.001% by weight or more, a more excellent effect can be obtained. In addition, the proportion of component (B) to the total amount of the composition for external use E is preferably 10% by weight or less, more preferably 7% by weight or less, and even more preferably 5% by weight or less. Within the above range, the topical composition E can have a good feel when used. By setting the ratio of the amount of component (B) to the total amount of the topical composition E to 10% by weight or less, a better feel when used can be obtained. The content of component (B) is, for example, 0.0001 to 25 parts by weight per part by weight of component (A). Component (B) is preferably contained in an amount of 0.01 to 10 parts by weight, more preferably 0.05 to 5 parts by weight, and even more preferably 0.1 to 2 parts by weight per part by weight of component (A).

[0034] Component (C) is a nonionic surfactant. Component (C) may be used alone or in any combination of two or more kinds.

[0035] Examples of the nonionic surfactant include ester-based nonionic surfactants, ether-based nonionic surfactants, alkyl glycosides, and silicone-based surfactants.

[0036] Examples of ester-based nonionic surfactants include sorbitan fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, polyglycerin fatty acids, propylene glycol fatty acid esters, hydrogenated castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, and polyoxyethylene monococonut fatty acid glyceryl.

[0037] Examples of sorbitan fatty acid esters include nonionic surfactants such as sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, diglycerol sorbitan penta-2-ethylhexyl acid, and diglycerol sorbitan tetra-2-ethylhexyl acid. Examples of glycerin fatty acid esters include glyceryl monostearate, glyceryl monostearate malic acid, and the like. Examples of polyglycerin fatty acid esters include polyglyceryl monostearate, polyglyceryl monoisostearate, polyglyceryl diisostearate, polyglyceryl monolaurate, polyglyceryl monooleate, and polyglyceryl monomyristate. Examples of polyglycerin fatty acids include polyglyceryl monostearate, polyglyceryl monoisostearate, polyglyceryl diisostearate, polyglyceryl monolaurate, polyglyceryl monooleate, and glycerin fatty acid esters. Examples of propylene glycol fatty acid esters include propylene glycol monostearate and polyethylene glycol fatty acid esters. Examples of hydrogenated castor oil derivatives include polyoxyethylene hydrogenated castor oils such as polyoxyethylene hydrogenated castor oil 40 (HCO-40), polyoxyethylene hydrogenated castor oil 50 (HCO-50), polyoxyethylene hydrogenated castor oil 60 (HCO-60), and polyoxyethylene hydrogenated castor oil 80. Examples of polyoxyethylene sorbitan fatty acid esters include polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan monostearate (polysorbate 60), polyoxyethylene (20) sorbitan monooleate (polysorbate 80), and polyoxyethylene (20) sorbitan isostearate.

[0038] Examples of the ether-based nonionic surfactant include glycerin alkyl ether, polyoxyalkylene alkyl ether, and polyoxyethylene polycyclic phenyl ether.

[0039] Examples of polyoxyalkylene alkyl ethers include polyoxyethylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene lauryl ether, polyoxyethylene cetyl ether, polyoxyethylene oleyl ether, polyoxyethylene tridecyl ether, polyoxyethylene castor oil ether, polyoxyethylene-2-ethylhexyl ether, polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, and polyoxyethylene behenyl ether.

[0040] Examples of the alkyl glycoside include octyl glycoside, decyl glycoside, and lauryl glycoside.

[0041] Examples of silicone surfactants include polyoxyethylene-methylpolysiloxane copolymer, lauryl PEG-9 polydimethylsiloxyethyl dimethicone, and PEG-9 polydimethylsiloxyethyl dimethicone.

[0042] The HLB value of the nonionic surfactant is based on the value measured by the Kawakami method. The HLB value is preferably 8 or more, particularly 9 or more, and more preferably 10 or more, in order to improve the compatibility of the composition with the skin. In other words, it is preferable that the nonionic surfactant is not hydrophobic.

[0043] In the topical composition E, the proportion of the amount of component (C) (when two or more components are used as component (C), the sum of the amounts of the components) in the total amount of the topical composition E is preferably 0.0001% by weight or more, more preferably 0.005% by weight or more, even more preferably 0.01% by weight or more, and particularly preferably 0.05% by weight or more. By making the proportion of the amount of component (C) in the total amount of the topical composition E 0.0001% by weight or more, more excellent effects can be obtained. In addition, the proportion of component (C) in the total amount of the topical composition E is preferably 25% by weight or less, more preferably 10% by weight or less, and even more preferably 5% by weight or less. By making the proportion of the amount of component (C) in the total amount of the topical composition E 25% by weight or less, the sticky feeling during use of the topical composition E is suppressed, and a more excellent feeling of use can be obtained. The content of component (C) is, for example, 0.1 to 80 parts by weight relative to 1 part by weight of component (A). Component (C) is preferably contained in an amount of 1 to 50 parts by weight, more preferably 5 to 40 parts by weight, and even more preferably 6 to 35 parts by weight, relative to 1 part by weight of component (A).

[0044] Composition E for external use, which contains the above-mentioned components (A), (B), and (C), can suppress, prevent, prevent, or improve pigmentation of the skin after scratching an affected area in cases of skin diseases accompanied by itching, for example. In addition, composition E for external use, which contains the above-mentioned components (A), (B), and (C), can promote disappearance and / or fading of hematomas occurring due to bruises, etc.

[0045] In order to further enhance the effectiveness against various skin diseases, in addition to the above essential components, various active ingredients such as nonsteroidal anti-inflammatory agents, vitamins, anti-inflammatory analgesics, antifungal agents, steroids, local anesthetics, antibacterial agents, antiviral agents, keratin softeners, moisturizers, astringents, antioxidants, and cooling agents can be blended into the external composition E as necessary. The amounts of these ingredients to be blended are not particularly limited as long as the effects of the present invention are achieved, but are preferably selected appropriately within the upper limit of the pharma- ceutical acceptable amounts.

[0046] In addition to the above-mentioned active ingredients, topical composition E may contain water-soluble ingredients, preservatives, pH adjusters, thickeners, antioxidants, etc., which are typically contained in topical compositions, as necessary.

[0047] Examples of the dosage form of the external composition E include ointments, creams, gels, emulsions, external liquids, patches, solids, poultices, packs, tapes, and the like.

[0048] The topical composition E can be prepared by a conventional method for each formulation. At this time, known excipients and other ingredients necessary for preparing formulations of various dosage forms are appropriately blended.

[0049] The amount of composition E to be applied to the skin and the method of use are not particularly limited, and for example, an appropriate amount (e.g., about 0.05 to 5 g) may be applied to the skin several times a day (e.g., about 1 to 5 times, preferably 1 to 3 times). The amount of composition E to be applied to the skin and the method of use vary depending on the skin condition, age, sex, etc. of the subject. The application period may be, for example, about 2 weeks to 6 months, preferably about 1 to 6 months. Composition E for external use can be suitably used for people who are at risk of developing skin pigmentation due to melanin production promoted by itching, or people who have skin pigmentation caused by melanin production promoted by itching.

[0050] The topical composition E according to the present embodiment has been described as being used as a skin drug to be applied to areas affected by dry skin diseases such as senile xerosis, atopic dermatitis, and housewives' eczema, inflammatory skin diseases, skin pruritus, heat rash, chilblains, cracks, and chapped skin, but the use of the topical composition E is not particularly limited thereto. For example, the topical composition E may be used as a skin drug to be applied to pigmentation (e.g., scars) that occurs on the skin after external wounds such as burns and trauma have healed completely. The topical composition E may also be used as a skin drug to be applied to pigmentation that occurs due to, for example, hormone imbalance, aging, sudden strong sunburn, chronic friction, etc. Furthermore, composition E for external use may be applied to the skin for the purpose of preventing or preventing pigmentation even before pigmentation occurs, after a burn or wound from an external injury has completely healed, or after the occurrence of hormone imbalance, aging, sudden strong sunburn, chronic friction, etc. Composition E for external use may also be applied to an area of ​​the skin where symptoms of bruises (e.g., black bruises, brown bruises, blue bruises) are observed, or composition E for external use may be applied for the purpose of preventing or preventing bruises. EXAMPLES

[0051] The present invention will be specifically described below with examples and comparative examples prepared in Evaluation Test I and Evaluation Test II, but the present invention is not limited to the following examples. In this experiment, a cell lysate containing melanoma cells was prepared, and the amount of melanin produced per cell was measured to evaluate the effect of reducing the amount of melanin produced, which was used as an index of the effect of suppressing, preventing, preventing, or improving pigmentation. Note that the present invention evaluates pigmentation from the perspective of suppressing melanin production.

[0052] (Evaluation Test I) (Sample preparation) First, mouse-derived B16 melanoma cells were seeded in 3 mL of DMEM (Dulbecco's Modified Eagle) medium containing 10% FBS (fetal bovine serum) in a 6-well plate (15.53 mL per well). At this time, the cell density of mouse-derived B16 melanoma cells was 3.0 × 10 4The plate was then cultured at 37°C in an environment of 5% carbon dioxide and 95% air for 1 day.

[0053] After that, the medium was replaced with a new DMEM medium, and the test drug prepared using 10% FBS-containing DMEM medium was added to each well so that the final concentration was a predetermined value. The type and concentration of the test drug added here differed depending on each Example, Comparative Example, and Reference Example, which will be described later. Then, the sample was further cultured for 3 days. The final liquid volume in the plate was about 3 mL per well.

[0054] After incubation, the supernatant from each well was removed and the samples were washed once with PBS(-). Then, 600 μL of 0.1N NaOH was added to each well and left to stand at room temperature for 24 hours. This caused the cells in the plate to dissolve, and a cell lysate was obtained. In the cell lysate, the melanin pigment was completely dissolved by the alkaline treatment.

[0055] (Test Drug) In each of Examples 1 to 12 and Comparative Examples 2 to 17, the test drugs used were adjusted so that the final concentrations of components (A) to (C) contained in the cell lysate were the values ​​shown below. Note that no test drug was added in Comparative Example 1. The final concentrations of components (A) to (C) contained in each cell lysate are also shown in Tables 1 to 6.

[0056] Comparative Example 1 Comparative Example 1 was a test drug to which none of components (A) to (C) was added.

[0057] (Comparative Examples 2 to 3) As the test drug, only component (A) of components (A) to (C) was added, which were set as Comparative Examples 2 and 3. Comparative Examples 2 and 3 were prepared so that the final concentrations of the heparinoid were 0.006% by weight and 0.002% by weight, respectively.

[0058] (Comparative Examples 4 to 7) Comparative Examples 4 to 7 were prepared by adding only component (B) of components (A) to (C) as the test drug. Comparative Example 4 was prepared so that the final concentration of panthenol was 0.01% by weight. Comparative Examples 5, 6, and 7 were prepared so that the final concentrations of allantoin were 0.01% by weight, 0.02% by weight, and 0.1% by weight, respectively.

[0059] (Comparative Examples 8 to 11) Comparative Examples 8 to 11 were prepared by adding only component (C) out of components (A) to (C) as the test drug. Comparative Examples 8, 9, and 10 were prepared so that the final concentrations of polyoxyethylene hydrogenated castor oil 50 were 0.004% by weight, 0.01% by weight, and 0.04% by weight, respectively. Comparative Example 11 was prepared so that the final concentration of polysorbate 60 was 0.02% by weight.

[0060] (Comparative Examples 12 to 13) Comparative Examples 12 and 13 were prepared by adding only components (A) and (B) of components (A) to (C) as test drugs. Comparative Example 12 was prepared so that the final concentration of the heparinoid was 0.006% by weight, and the final concentration of panthenol was 0.01% by weight. Comparative Example 13 was prepared so that the final concentration of the heparinoid was 0.006% by weight, and the final concentration of allantoin was 0.02% by weight.

[0061] Comparative Example 14 The test drug was prepared as Comparative Example 14 by adding only components (A) and (C) out of components (A) to (C). Comparative Example 14 was prepared so that the final concentration of the heparinoid was 0.006% by weight, and the final concentration of polyoxyethylene hydrogenated castor oil 50 was 0.04% by weight.

[0062] (Comparative Examples 15 to 17) Comparative Examples 15 to 17 were prepared by adding only components (B) and (C) of components (A) to (C) as test drugs. Comparative Example 15 was prepared so that the final concentration of panthenol was 0.01% by weight, and the final concentration of polysorbate 60 was 0.02% by weight. Comparative Example 16 was prepared so that the final concentration of allantoin was 0.02% by weight, and the final concentration of polyoxyethylene hydrogenated castor oil 50 was 0.04% by weight. Comparative Example 17 was prepared so that the final concentration of allantoin was 0.1% by weight, and the final concentration of polyoxyethylene hydrogenated castor oil 50 was 0.04% by weight.

[0063] (Examples 1 to 4) Examples 1 to 4 were prepared by blending a heparinoid as the test drug for component (A), allantoin as the test drug for component (B), and polyoxyethylene hydrogenated castor oil 50 as the test drug for component (C). In Examples 1 to 4, the amount of the heparinoid was adjusted so that the final concentration was 0.006% by weight. The amount of allantoin was adjusted so that the final concentration was 0.02% by weight in Examples 1 to 3 and 0.1% by weight in Example 4. The ratio of the amount of component (B) to the amount of component (A) was 3.33 in Examples 1 to 3 and 16.7 in Example 4. The amount of polyoxyethylene hydrogenated castor oil 50 was adjusted so that the final concentrations in Examples 1, 2, 3, and 4 were 0.01% by weight, 0.04% by weight, 0.2% by weight, and 0.1% by weight, respectively. The ratio of the amount of component (C) to the amount of component (A) was 1.67 parts by weight, 6.67 parts by weight, 33.3 parts by weight, and 16.7 parts by weight in Examples 1 to 4, respectively.

[0064] (Examples 5 to 9) Examples 5 to 9 were prepared by blending a heparinoid as the test drug for component (A), panthenol as the test drug for component (B), and polyoxyethylene hydrogenated castor oil 50 as the test drug for component (C). The final concentration of the heparinoid was adjusted to 0.006% by weight in Examples 5 and 6, and 0.002% by weight in Examples 7 to 9. In Examples 5 to 9, the final concentration of panthenol was adjusted to 0.01% by weight. The ratio of the amount of component (B) to the amount of component (A) was 1.67 in Examples 5 and 6, and 5 in Examples 7 to 9. The amount of polyoxyethylene hydrogenated castor oil 50 was adjusted to 0.01% by weight, 0.04% by weight, 0.004% by weight, 0.01% by weight, and 0.04% by weight in Examples 5, 6, 7, 8, and 9, respectively. The ratio of the amount of component (C) to the amount of component (A) was 1.67 parts by weight, 6.67 parts by weight, 2 parts by weight, 5 parts by weight, and 20 parts by weight in Examples 5 to 9, respectively.

[0065] (Examples 10 to 12) A sample containing the same formulation as in Example 5 except that polysorbate 60 was used instead of polyoxyethylene hydrogenated castor oil 50 as component (C) and the final concentration was adjusted to 0.02% by weight was used as component (C), was designated as Example 10. The ratios of components (B) and (C) to the amount of component (A) were 1.67 and 3.33, respectively. In addition, a sample containing the same formulation as in Example 10 except that allantoin was used instead of panthenol as component (B) and the final concentration was adjusted to 0.01% by weight was used as component (B), was designated as Example 11. The ratios of components (B) and (C) to the amount of component (A) were 1.67 and 3.33, respectively. Furthermore, a sample containing the test drug in the same formulation as in Example 8, except that polysorbate 60 was used as component (C) instead of polyoxyethylene hydrogenated castor oil 50 and the final concentration was adjusted to 0.01% by weight, was prepared as Example 12. The ratios of components (B) and (C) to the amount of component (A) were 5 parts by weight and 5 parts by weight, respectively.

[0066] (Evaluation method) The cell lysates of the Examples and Comparative Examples were measured for absorbance at 475 nm using a spectrophotometer (Microplate Reader SH-9000Lab, manufactured by Corona Electric Co., Ltd.) to determine the melanin production level. The melanin production level here refers to the amount of melanin in all melanoma cells.

[0067] In addition, the cell lysates of the Examples and Comparative Examples were subjected to protein quantification using a BCA protein quantification kit (manufactured by Thermo Fisher Scientific Co., Ltd., BCA Protein Assay Reagent). Here, the protein amount of B16 melanoma cells was measured. This measurement value was used as an index of the number of melanoma cells in the entire cell lysate.

[0068] To evaluate the amount of melanin per melanoma cell, the amount of melanin produced was divided by the amount of protein, as shown in the following formula, and this value was used as the standard value for the amount of melanin. The standard value for the amount of melanin was evaluated as a relative value.

[0069] (Melanin amount standard value) = (Melanin production amount) / (protein amount) Specifically, for Comparative Examples 1 to 3, the melanin amount standard value of Comparative Example 1 was set as standard value m0 (100). In addition, the melanin amount standard value m of Comparative Examples 2 and 3 was divided by the melanin amount standard value m0 of Comparative Example 1, and the result was subtracted from 1 to express it as a percentage, which was set as the melanin production reduction rate r of Comparative Examples 2 and 3.

[0070] Regarding the melanin production reduction rate r in Examples 1 to 6, Examples 10 to 12, and Comparative Examples 4 to 17, the respective melanin amount standard values ​​m are calculated based on the melanin amount standard value m when only a heparinoid is added as the test drug to a final concentration of 0.006% by weight. 0.006 The percentage was calculated by dividing the amount of melanin produced by the test drug by the standard value m of the amount of melanin, subtracting the result from 1.0.002a The percentage is calculated by dividing the amount of melanin produced by the skin and subtracting the result from 1. The formula for calculating the rate of decrease in melanin production r is shown below.

[0071]

number

[0072] (Evaluation Results) The evaluation results are shown in Tables 1 to 6.

[0073] [Table 1]

[0074] [Table 2]

[0075] [Table 3]

[0076] [Table 4]

[0077] [Table 5]

[0078] [Table 6]

[0079] According to Table 1, comparing Comparative Example 1, which does not contain a heparinoid in the cell lysis solution, with Comparative Examples 2 and 3, which contain a heparinoid, Comparative Example 2, in which the final concentration of the heparinoid is 0.006% by weight, has a lower effect of reducing the amount of melanin produced than Comparative Example 1, while Comparative Example 3, in which the final concentration of the heparinoid is 0.002% by weight, has a slightly higher effect of reducing the amount of melanin produced than Comparative Example 1. This shows that even if a heparinoid is added alone to the cell lysis solution, it does not contribute to reducing melanin production.

[0080] According to Table 2, in Comparative Examples 4 to 7 in which component (B) allantoin or panthenol was blended in the cell lysis solution, the amount of melanin produced was increased compared to when the heparinoid was blended alone. Therefore, when allantoin or panthenol was blended alone in the cell lysis solution, no effect of reducing the amount of melanin produced was obtained.

[0081] Also, according to Table 3, in Comparative Examples 8 to 11 in which component (C) polyoxyethylene hydrogenated castor oil 50 or polysorbate 60 was blended in the cell lysis solution, the amount of melanin produced was increased compared to the case in which a heparinoid was blended alone. Therefore, it is clear that when polyoxyethylene hydrogenated castor oil 50 or polysorbate 60 is blended alone in the cell lysis solution, no effect of reducing the amount of melanin produced is obtained at all.

[0082] Therefore, it is clear that components (A), (B) and (C) do not have the effect of reducing melanin production by themselves.

[0083] According to Table 4, it can be seen that the amount of melanin production is increased in Comparative Examples 12 and 13, which contain components (A) and (B) but not (C), Comparative Example 14, which contains components (A) and (C) but not (B), and Comparative Examples 15 to 17, which contain components (B) and (C) but not (A), compared to the case where a heparinoid is contained alone. This shows that the combination of two of the components (A), (B), and (C) does not have the effect of reducing the amount of melanin production.

[0084] Tables 5 and 6 show that in Examples 1 to 12, in which all of components (A), (B), and (C) were blended in the cell lysis solution, the amount of melanin production was reduced compared to when the heparinoid was blended alone. Therefore, it is evident that the effect of reducing the amount of melanin production can only be obtained by combining both components (B) and (C) with component (A), which has no effect on suppressing melanin production.

[0085] In addition, JP 2011-231128 A discloses that a skin preparation contains a heparinoid, allantoin, and pantothenic acid, and that a surfactant such as sorbitan monostearate is added as an additive. In addition, JP 2012-36183 A discloses a composition for external use containing a heparinoid, allantoin, and pantothenic acid, and describes that this composition for external use may contain allantoin, pantothenic acid, and a surfactant as optional components. In other words, it is known that all of the components (A), (B), and (C) are added to a composition for external use as in the present invention. However, these documents do not disclose anything about the effect of suppressing melanin production by combining the components (A), (B), and (C), and the effect of suppressing melanin production was discovered for the first time by the present inventors.

[0086] It is understood that the topical composition contains components (A), (B) and (C) and has the effect of reducing the amount of melanin produced in melanocytes, thereby suppressing or preventing skin pigmentation. It is also understood that the topical composition contains components (A), (B) and (C) and reduces the amount of melanin produced, so that when the keratinocytes in the basal layer of the skin turn over, the amount of melanin produced in new cells is reduced, thereby preventing or improving skin pigmentation. It is also understood that the topical composition contains components (A), (B) and (C) and has the effect of reducing the amount of melanin produced in melanocytes, so that in the dermis, it is also effective in preventing or improving pigmentation of bruises (e.g., black bruises, brown bruises, blue bruises).

[0087] Next, consider Examples 1 to 3 and 5 to 9 in which polyoxyethylene hydrogenated castor oil 50 is blended as component (C). FIG. 1 is a graph showing the relationship between the amount of polyoxyethylene hydrogenated castor oil 50 blended and the melanin production reduction rate. Here, E1 to E3 and E5 to E9 in the graph indicate the data of the melanin production reduction rate of Examples 1 to 3 and Examples 5 to 9, respectively. A comparison of Examples 1 to 3 in which a heparinoid (final concentration: 0.006% by weight) was blended as component (A), allantoin (final concentration: 0.02% by weight) was blended as component (B), and only the amount of component (C) was changed is shown by the broken line L1. In addition, a comparison of Examples 5 and 6 in which a heparinoid (final concentration: 0.006% by weight) was blended as component (A), panthenol (final concentration: 0.01% by weight) was blended as component (B), and only the amount of component (C) was changed is shown by the broken line L2. Furthermore, a solid line L3 shows a comparison of Examples 7 to 9 in which a heparinoid (final concentration: 0.002 wt%) was blended as component (A), panthenol (final concentration: 0.01 wt%) was blended as component (B), and only the amount of component (C) was changed.

[0088] From the graph of FIG. 1, it can be seen that, for all of the broken line L1, the broken line L2, and the solid line L3, when the amount of polyoxyethylene hydrogenated castor oil 50 in the cell lysate is 0.04% by weight, the melanin production reduction rate increases to nearly 40%. Also, for the broken line L1, when the amount of polyoxyethylene hydrogenated castor oil 50 increases beyond 0.04% by weight, the melanin production reduction rate becomes constant at about 40%. As a result, it can be seen that the change in the melanin production reduction rate changes significantly around the amount of polyoxyethylene hydrogenated castor oil 50 of 0.04% by weight. Therefore, the amount of polyoxyethylene hydrogenated castor oil 50 of 0.04% by weight is an inflection point for the change in the melanin production reduction rate, and has a critical significance in increasing the melanin production reduction rate to about 40%. Therefore, it is found that an excellent melanin production reduction rate can be obtained when the amount of component (C) is 0.04% by weight or more, regardless of the type and amount of component (A) and component (B). For this reason, it is more preferable that the lower limit of the amount of polyoxyethylene hydrogenated castor oil 50 is 0.04% by weight or more.

[0089] From this, it is believed that even in topical compositions for actual use, a particularly excellent effect of reducing melanin production can be obtained when the amount of component (C) is a predetermined concentration or higher, regardless of the amounts and types of components (A) and (B).

[0090] (Evaluation Test II) Next, Example 13 and Comparative Examples 18 to 19 prepared in Evaluation Test II will be described.

[0091] (Sample preparation) Samples were prepared in the same manner as in Evaluation Test I, except that in the melanoma cell dissolution treatment, the concentration of NaOH added to each well was 4N and the dissolution treatment conditions were 60°C and 3 hours.

[0092] (Test Drug) The test drugs used in Example 13 and Comparative Examples 18 to 19 were adjusted so that the final concentrations of components (A) to (C) contained in the cell lysate were as shown below. The final concentrations of components (A) to (C) contained in each cell lysate are also shown in Table 7.

[0093] (Comparative Examples 18 and 19) Comparative Example 18 was prepared by adding only component (A) of components (A) to (C) as the test drug. Comparative Example 18 was prepared so that the final concentration of the heparinoid was 0.002% by weight. Comparative Example 19 was prepared by adding only component (B) of components (A) to (C) as the test drug. Comparative Example 19 was prepared so that the final concentration of vitamin A oil was 0.003% by weight.

[0094] (Example 13) Example 13 was prepared by blending a heparinoid as the test drug for component (A), vitamin A oil as the test drug for component (B), and polyoxyethylene hydrogenated castor oil 40 as the test drug for component (C). In Example 13, the blending amounts were adjusted so that the final concentration of the heparinoid was 0.002% by weight, the final concentration of the vitamin A oil was 0.003% by weight, and the final concentration of the polyoxyethylene hydrogenated castor oil 40 was 0.16% by weight. The ratio of the blending amount of component (B) to the blending amount of component (A) was 1.5 parts by weight. The ratio of the blending amount of component (C) to the blending amount of component (A) was 80 parts by weight.

[0095] (Evaluation method) For the cell lysates of Example 13 and Comparative Examples 18 to 19, absorbance was measured and protein was quantified in the same manner as in Evaluation Test I. From these results, a melanin amount standard value was obtained based on the following formula, as in Evaluation Test I.

[0096] (Melanin amount standard value) = (Melanin production amount) / (protein amount)

[0097] The melanin amount standard value of Comparative Example 18 is m 0.002bIn addition, the melanin amount standard value of Example 13 or Comparative Example 19 and the melanin amount standard value m 0.002b The melanin production reduction rate r of Example 13 or Comparative Example 19 was calculated using the above.

[0098]

number

[0099] (Evaluation Results) The results of these evaluations are shown in Table 7.

[0100] [Table 7]

[0101] According to Table 7, it was found that even when vitamin A oil was added as component (B), the effect of reducing melanin production was observed when used in combination with components (A) and (C). [Industrial Applicability]

[0102] The present invention is useful for a composition for external use used for suppressing, preventing, prophylaxis or amelioration of skin diseases.

Claims

[Claim 1] A composition for external use used for suppressing, preventing, preventing or improving a skin disease, comprising: (A) a heparinoid; (B) one or more selected from the group consisting of allantoin, pantothenic acid, vitamin A, pharma- ceutically acceptable salts thereof, and pharma-ceutically acceptable derivatives thereof; (C) a nonionic surfactant; and A topical composition comprising:

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