JP2004231558A - External preparation for skin for bleaching - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an external preparation for skin for bleaching containing an extract of stamen of Nelumbo nucifera Gaertn. <P>SOLUTION: The external preparation for skin for bleaching comprises the extract of stamen of Nelumbo nucifera Gaertn. The external preparation for skin for bleaching comprising the extract of stamen of Nelumbo nucifera Gaertn exhibits excellently bleaching action on the skin. <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

【００２８】
実験例２ メラニン生成抑制試験
Ｂ−１６マウスメラノーマ細胞におけるメラニン生成抑制効果を下記の条件にて測定した。
【００２９】
Ｂ−１６マウスメラノーマ細胞をφ６０ｍｍのｄｉｓｈに１×１０^５個播種し、１００、１０００μｇ／ｍＬの試料を添加したＥａｇｌｅ’ｓ ＭＥＭ培地にて３７℃、５％ＣＯ_２の条件下で３日間培養した。次に細胞をｄｉｓｈから剥離し、細胞を超音波破砕した後、４Ｎ ＮａＯＨを加え６０℃で２時間加温することによりメラニンを溶解させ、分光光度計で４７５ｎｍにおける吸光度を測定した。なお、超音波処理後の細胞破砕液中のタンパク量を測定し、タンパク量当りのメラニン量によって、メラニン生成量とした。コントロールのメラニン生成量に対する試料添加時のメラニン生成量値からメラニン生成抑制率を求めた。
【００３０】
これらの試験結果を表２に示した。その結果、蓮鬚の抽出物には優れたメラニン生成抑制効果が認められた。
【００３１】
【表２】

【００３２】
実験例３ ＩＬ−１α生成抑制試験
培養ケラチノサイトにおけるＵＶＢ照射後のＩＬ−１α生成促進に対する抑制効果を下記の条件にて測定した。
【００３３】
コンフルエントな状態のＰａｍ２１２細胞に２０ｍＪ／ｃｍ^２のＵＶＢを照射した後、１０μｇ／ｍＬの試料を添加したＥａｇｌｅ’ｓ ＭＥＭ培地にて３７℃、５％ＣＯ_２の条件下で、さらに２４時間培養した。その後、培養上清を回収し、Ｍｏｕｓｅ ＩＬ−１α ＥＬＩＳＡ ｋｉｔ（Ｅｎｄｏｇｅｎ社）にてＩＬ−１α濃度を測定した。コントロールのＩＬ−１α生成量に対する試料添加時のＩＬ−１α生成量値からＩＬ−１α生成抑制率を求めた。
【００３４】
これらの試験結果を表３に示した。その結果、蓮鬚の抽出物には優れたＩＬ−１α生成抑制効果が認められた。
【００３５】
【表３】

【００３６】
実験例４ 使用試験
実施例１のクリーム及び比較例１の従来のクリームを用いて、各々女性３０人（３０〜４５才）を対象に１ヶ月間の使用試験を行った。使用後、シミ、ソバカス及び透明感の改善についてのアンケート調査を行って、美白効果を判定した。アンケートの評価基準は、有効なものを「優」、やや有効なものを「良」、わずかに有効なものを「可」、無効なものを「不可」として評価した。
【００３７】
これらの結果を表４に示した。実施例１の蓮鬚熱水抽出物を含有することを特徴とする皮膚外用剤は優れた美白効果を示した。なお、試験期間中皮膚トラブルは一人もなく、安全性においても問題なかった。
【００３８】
【表４】

【００３９】
実施例２の化粧水、実施例３の乳液、実施例４の軟膏、実施例５のファンデーション及び実施例６の浴用剤の使用試験を行ったところ、いずれも安全で優れた美白効果を示した。
【００４０】
【発明の効果】
以上のことから、本発明の蓮鬚の抽出物は優れたメラノサイトの活性化抑制効果、メラニン生成抑制効果及びＩＬ−１α生成抑制効果を示した。蓮鬚の抽出物を含有することを特徴とする美白用皮膚外用剤は安全で優れた皮膚の美白作用を示した。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a skin external preparation having an excellent whitening effect.
[0002]
[Prior art]
Patent Document 1 is known as a skin external preparation characterized by containing a beard extract, but it has not been known that this extract has a whitening effect.
Conventionally, it has been considered that the production of melanin is excessive in melanocytes as a cause of the stain, and suppression of melanin production in melanocytes has been a theme of the development of whitening agents.
In recent years, as one of the melanocyte activation mechanisms, in keratinocytes irradiated with ultraviolet light, first, the production of interleukin-1α (IL-1α) is enhanced, and this IL-1α stimulates keratinocytes themselves, and endothelin and prostasin from keratinocytes Promotes the production of melanocyte activators such as glandins. It has been revealed that these melanocyte activating factors stimulate melanocytes to cause melanocyte activation (see Non-Patent Documents 1 and 2). Therefore, various components that suppress the activation of melanocytes by these melanocyte activating factors have been developed as whitening agents.
[0003]
[Patent Document 1]
JP-A-11-71234 [Non-Patent Document 1]
Imokawa, G .; , J. et al. Biol. Chem. , 1992, 267, 24675-24680
[Non-patent document 2]
Nordlund, J .; J. , J. et al. Invest. Dermatol. , 1986, 86, 433-437.
[0004]
[Problems to be solved by the invention]
There are various factors involved in activation of melanocytes by ultraviolet irradiation, and it is not efficient to develop a component that suppresses the action or production of each of them. In UV-irradiated keratinocytes, first, IL-1α production is enhanced, and since this IL-1α promotes the production of melanocyte activator, it is possible to suppress IL-1α production from keratinocytes by UV irradiation. It is considered that the activation of melanocytes is efficiently suppressed, leading to an excellent whitening effect.
[0005]
[Means for Solving the Problems]
Under such circumstances, the present inventors have conducted intensive studies and found that the extract of lotus bean has an excellent inhibitory effect on the promotion of IL-1α production from keratinocytes and activation of melanocytes by ultraviolet irradiation. The inventors have found that the present invention has, and have completed the present invention.
[0006]
That is, the present invention is an external preparation for whitening skin, characterized by containing an extract of lotus beard.
[0007]
The beard used in the present invention is a stamen of a lotus (Nelumbo nucifera Gaertn.) Of the Nymphaeaaceae family. Lotus is distributed on the Eurasian continent and North America, but the main source of lotus is in China's Zhejiang and Jiangsu provinces. Although lotus is grown in Japan, lotus roots are used only for food, and lotus is not collected. Renge is commercially available and its hot water extract is known as astringent, hemostatic and tonic.
[0008]
The extract of the beard used in the present invention is obtained by immersing or heating the beard together with the extraction solvent, followed by filtration and, if necessary, concentration. Examples of the extraction solvent include water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol and the like), liquid polyhydric alcohols (1,3-butylene glycol, propylene) Glycols), hydrocarbons (benzene, hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.), acetonitrile and the like. These solvents may be used alone or as a mixture of two or more. Preferably, one or more of water or a water-soluble solvent (a solvent that can be mixed with water at an arbitrary ratio, for example, ethanol, 1,3-butylene glycol, propylene glycol, and the like) may be used. . The extract may be used as it is, or part or all of the solvent may be distilled off.
[0009]
In the skin whitening preparation for external use of the present invention, the above extract of lotus bean may be used as it is, and as long as the effect is not impaired, oils and fats, waxes, and carbonized ingredients which are commonly used in external preparations are used. Hydrogens, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, humectants, powders, ultraviolet absorbers, thickeners, pigments, antioxidants, whitening agents Also, components such as a chelating agent can be blended.
[0010]
The extract of beard used in the present invention can be used in any of cosmetics, quasi-drugs, and pharmaceuticals. Examples of the dosage form include lotions, creams, emulsions, gels, aerosols, and patches. Preparations, essences, packs, detergents, bath preparations, foundations, powders, lipsticks, ointments, shampoos, rinses, treatments, tonics and the like.
[0011]
The amount of the extract of the mustard extract used in the present invention is not particularly limited, but is preferably in the range of 0.0001 to 75% by weight, more preferably 0.001 to 30% by weight as a dry matter. If it is less than 0.0001% by weight, the whitening effect is low, and if it exceeds 75% by weight, the effect is hardly greatly enhanced and it is not efficient. The method of addition may be added in advance or during the production, and may be appropriately selected in consideration of workability.
[0012]
Next, in order to explain the present invention in detail, Production Examples, Formulation Examples, and Experimental Examples of the extract used in the present invention will be given as Examples, but the present invention is not limited thereto. The blending amounts shown in the examples represent% by weight.
[0013]
Production Example 1 Lotus beard hot water extract 1 L of water was added to 50 g of dried beard and extracted at 95 to 100 ° C. for 2 hours, insolubles were filtered, and the filtrate was concentrated and dried to dryness. 12 g of beard hot water extract was obtained.
[0014]
Production Example 2 Ethanol extract of beard 10 L of ethanol was added to 500 g of dried beard and extracted at room temperature for 7 days. The insoluble material was filtered, and the filtrate was concentrated to dryness to obtain 80 g of an ethanol extract of beard. Obtained.
[0015]
Production Example 3 300 g of 1,3-butylene glycol extract was added to 30 g of dried beard and extracted at normal temperature for 10 days. -280 mL of butylene glycol extract was obtained.
[0016]
Example 1 Cream Formulation Amount (% by weight)
1. Renge hot water extract (Production Example 1) 0.05
2. Squalane 5.5
3. Olive oil 3.0
4. Stearic acid 2.0
5. Beeswax 2.0
6. Octyldodecyl myristate 3.5
7. Polyoxyethylene cetyl ether (20EO) 3.0
8. Behenyl alcohol 1.5
9. Glycerin monostearate 2.5
10. Fragrance 0.1
11.1,3-butylene glycol 8.5
12. Ethyl paraoxybenzoate 0.05
13. Methyl paraoxybenzoate 0.2
14． Purified water 68.1
[Production method] Components 2 to 9 are mixed by heating, and kept at 70 ° C to obtain an oil phase. Components 1 and 11 to 14 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase, emulsified, cooled while stirring, and the component 10 is added at 45 ° C, and further cooled to 30 ° C to obtain a product.
[0017]
Comparative Example 1 Conventional cream A conventional cream was prepared by replacing the lotus bean hot water extract with purified water in Example 1.
[0018]
Example 2 Lotion Formulation Amount (% by weight)
1. Lotus beard ethanol extract (Production Example 2) 0.01
2.1,3-butylene glycol 8.0
3. Glycerin 2.0
4. Xanthan gum 0.02
5. Citric acid 0.01
6. Sodium citrate 0.1
7. Ethanol 5.0
8. Methyl paraoxybenzoate 0.1
9. Polyoxyethylene hydrogenated castor oil (40EO) 0.1
10. Fragrance 0.1
11. Purified water 84.56
[Manufacturing method] Components 1 to 6 and 11 and components 7 to 10 are each uniformly dissolved, and both are mixed and filtered to obtain a product.
[0019]
Example 3 Emulsion Formulation Amount (% by weight)
1. Renge 1,3-butylene glycol extract (Production Example 3) 0.05
2. Squalane 5.0
3. Olive oil 5.0
4. Jojoba oil 5.0
5. Cetanol 1.5
6. Glycerin monostearate 2.0
7. Polyoxyethylene cetyl ether (20EO) 3.0
8. Polyoxyethylene sorbitan monooleate 2.0
9. Fragrance 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12. Methyl paraoxybenzoate 0.2
13. Purified water 73.15
[Production method] Components 2 to 8 are dissolved by heating and mixed, and the mixture is kept at 70 ° C to obtain an oil phase. Components 1 and 10 to 13 are dissolved by heating and mixed, and kept at 75 ° C. to obtain an aqueous phase. The oil phase is added to the water phase, emulsified, cooled while stirring, and the component 9 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.
[0020]
Example 4 Ointment Formulation Amount (% by weight)
1. Renge hot water extract (Production Example 1) 1.0
2. Polyoxyethylene cetyl ether (30EO) 2.0
3. Glycerin monostearate 10.0
4. Liquid paraffin 5.0
5. Cetanol 6.0
6. Methyl paraoxybenzoate 0.1
7. Propylene glycol 10.0
8. Purified water 65.9
[Production method] Components 2 to 5 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Components 1 and 6 to 8 are dissolved by heating and mixed, and the mixture is kept at 75 ° C. to form an aqueous phase. The oil phase is added to the water phase, emulsified, and cooled to 30 ° C. with stirring to obtain a product.
[0021]
Example 5 Foundation Formulation Amount (% by weight)
1. Lotus beard ethanol extract (Production Example 2) 1.0
2. Stearic acid 2.4
3. Polyoxyethylene sorbitan monostearate 1.0
(20EO)
4. Polyoxyethylene cetyl ether (20EO) 2.0
5. Cetanol 1.0
6. Liquid lanolin 2.0
7. Liquid paraffin 3.0
8. Isopropyl myristate 6.5
9. Butyl paraoxybenzoate 0.1
10. Sodium carboxymethyl cellulose 0.1
11. Bentonite 0.5
12. Propylene glycol 4.0
13. Triethanolamine 1.1
14． Methyl paraoxybenzoate 0.2
15. Titanium dioxide 8.0
16. Talc 4.0
17. Bengala 1.0
18. Yellow iron oxide 2.0
19. Fragrance 0.1
20. Purified water 60.0
[Production method] Components 2 to 9 are heated and dissolved, and kept at 80 ° C to obtain an oil phase. Swell component 10 well with component 20, then add components 1 and 11-14 and mix uniformly. Components 15 to 18 crushed and mixed by a crusher are added thereto, and the mixture is stirred with a homomixer and kept at 75 ° C. to obtain an aqueous phase. The oil phase is added to the aqueous phase while stirring, cooled, and the component 19 is added at 45 ° C, and cooled to 30 ° C with stirring to obtain a product.
[0022]
Example 6 Formulation of bath agent Formulation amount (% by weight)
1. Renge 1,3-butylene glycol extract (Production Example 3) 5.0
2. Sodium bicarbonate 50.0
3. Yellow 202 (1) 0.05
4. Fragrance 0.25
5. Anhydrous sodium sulfate 44.7
[Production method] Components 1 to 5 are uniformly mixed to obtain a product.
[0023]
Next, experimental examples will be described in order to explain the effects of the present invention in detail.
[0024]
Experimental Example 1 Melanocyte Activation Inhibition Test Melanocyte activation inhibition effect was measured under the following conditions.
[0025]
Auricular skin of C57BL / 6 mice (male, 6 weeks old, 4 mice per group) was irradiated with 120 mJ / cm ² of UVB, and immediately and 6 hours later, the auricular skin was washed with ethanol, and then a 50 μL sample (20% ethanol solution containing 1% of Production Examples 1 to 3) was applied. After washing the auricle skin with ethanol for the next 12 days, 50 μL of the sample was applied twice a day. One day after the final sample application, the auricle skin was collected and incubated at 37 ° C for 2 hours in a 2N sodium bromide solution to prepare an epidermal sheet, and then at 37 ° C in a 0.1% L-DOPA solution. Dopa staining was performed by incubating for 3 hours. The number of melanocytes that became positive by dopa staining (dopa-positive melanocytes) was counted for 10 fields per sample at 100 times magnification of an optical microscope, and the average cell number was defined as the number of dopa-positive melanocytes in the sample. Similarly, auricle skin irradiated with UVB and coated with only the base was used as a control, and the activation inhibition rate of melanocytes was determined from the dopa-positive melanocyte value at the time of sample application relative to the control dopa-positive melanocyte count.
[0026]
The test results are shown in Table 1. As a result, the extract of lotus beard was found to have an excellent melanocyte activation inhibitory effect.
[0027]
[Table 1]

[0028]
Experimental Example 2 Melanin Production Inhibition Test B-16 The melanin production inhibitory effect on mouse melanoma cells was measured under the following conditions.
[0029]
1 × 10 ⁵ B-16 mouse melanoma cells were seeded on a φ60 mm dish, and cultured in Eagle's MEM medium supplemented with 100 and 1000 μg / mL samples at 37 ° C. and 5% CO ₂ for 3 days. did. Next, the cells were detached from the dish, the cells were ultrasonically crushed, melanin was dissolved by adding 4N NaOH and heating at 60 ° C. for 2 hours, and the absorbance at 475 nm was measured with a spectrophotometer. In addition, the amount of protein in the cell lysate after the ultrasonic treatment was measured, and the amount of melanin was determined based on the amount of melanin per protein amount. The melanin production inhibition rate was determined from the melanin production value at the time of sample addition relative to the control melanin production amount.
[0030]
Table 2 shows the test results. As a result, an excellent melanin production inhibitory effect was observed in the extract of lotus beard.
[0031]
[Table 2]

[0032]
Experimental Example 3 IL-1α Production Inhibition Test The inhibitory effect on the promotion of IL-1α production in cultured keratinocytes after UVB irradiation was measured under the following conditions.
[0033]
After irradiating the confluent Pam212 cells with UVB at 20 mJ / cm ² , the cells were further cultured in Eagle's MEM medium supplemented with a 10 μg / mL sample at 37 ° C. and 5% CO ₂ for 24 hours. . Thereafter, the culture supernatant was collected, and the IL-1α concentration was measured using a Mouse IL-1α ELISA kit (Endogen). The IL-1α production inhibition rate was determined from the IL-1α production amount at the time of sample addition relative to the control IL-1α production amount.
[0034]
Table 3 shows the test results. As a result, an excellent inhibitory effect on IL-1α production was observed in the extract of lotus beard.
[0035]
[Table 3]

[0036]
Experimental Example 4 Use Test Using the cream of Example 1 and the conventional cream of Comparative Example 1, a use test for 30 women (30 to 45 years old) was performed for one month. After use, a questionnaire survey on the improvement of spots, freckles and transparency was performed to determine the whitening effect. The evaluation criteria of the questionnaire were evaluated as "excellent" for valid, "good" for slightly effective, "acceptable" for slightly effective, and "impossible" for invalid.
[0037]
Table 4 shows the results. The topical skin preparation containing the hot water extract of lotus root of Example 1 showed an excellent whitening effect. There was no skin trouble during the test, and there was no problem in safety.
[0038]
[Table 4]

[0039]
When use tests of the lotion of Example 2, the emulsion of Example 3, the ointment of Example 4, the foundation of Example 5, and the bath agent of Example 6 were carried out, all showed a safe and excellent whitening effect. .
[0040]
【The invention's effect】
From the above, the extract of lotus beard of the present invention showed excellent melanocyte activation inhibitory effect, melanin production inhibitory effect and IL-1α production inhibitory effect. The skin whitening preparation for external use characterized by containing a beard extract showed safe and excellent skin whitening action.

Claims (3)

An external preparation for skin whitening, comprising an extract of lotus beard. A whitening agent characterized by containing a bean extract. An interleukin-1α production inhibitor comprising a beard extract.