JP3597067B2 - Antioxidant and composition containing the same - Google Patents

Description

上記混合物を３７℃、５分保温後、１００ｕｌの１ｍＭＮＡＤＨを添加し３７℃，１５分反応させ、９０ｕｌ，２％ＢＨＴと２ｍｌＴＣＡ−ＴＢＡ−ＨＣｌ溶液を添加混合後１００℃，１５分間加熱した。室温に冷却後、５０００ｒｐｍで１０分間遠心分離を行い、上清中の吸光度（５３５ｎｍ）を測定した。
結果を表１（第１表）に示す。抽出例１，２，３のティリア・メキシカーナ（Ｔｉｌｉａ ｍｅｘｉｃａｎａ）抽出物は、既に抗酸化力のあることで知られているｄｌ―α―トコフェロール、カテキンに匹敵するＤＰＰＨラジカル消去作用及びミトコンドリアにおける脂質過酸化の抑制効果を有することがわかった。
【００２７】
【表１】

【００２８】
［実施例１］（クリーム）
下記記載の配合量においてＢ成分をＡ成分に混合し、均一に加熱溶解して温度を８０℃にした。次いでＣ成分を注入撹拌混合した後、撹拌しながら３０℃まで冷却しクリームを得た。

【００２９】
［比較例１］（クリーム）
実施例１において成分（Ｂ）ティリア・メキシカーナ（Ｔｉｌｉａ ｍｅｘｉｃａｎａ）抽出物を除いた以外はすべて実施例１と同様にして調製し、前述した各試験に使用した。
【００３０】
［実施例２］（二相型ローション）
下記記載の配合量においてＡ成分を室温にて均一に混合溶解し、Ｂ成分をゆっくり撹拌添加し二相型ローションを得た。

【００３１】
［比較例２］（二相型ローション）
実施例２において成分（Ｂ）ティリア・メキシカーナ（Ｔｉｌｉａ ｍｅｘｉｃａｎａ）抽出物を除いた以外はすべて実施例２と同様にして調製し、前述した各試験に使用した。
【００３２】
［試験例２］
実施例１、２比較例１、２において得られた組成物の効果を以下の有用性評価試験の試験方法により評価した。
（１） 有用性評価試験
健康な女性（２５〜４０才）８０名を２０名ずつに４群に分け、それぞれ実施例１、２及び比較例１，２の試料を１日２回ずつ塗布し、塗布開始後３ヶ月後の老化防止効果（肌荒れ防止、皮膚の艶・張り）についてアンケート調査を行って評価した。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、比較例と比較して評価を行った。
第２表（表２）に示すごとく、比較例１、２に比べ実施例１、２では良好な結果が得られた。
【００３３】
【表２】

【００３４】
［実施例３］（油性軟膏）
下記記載の配合量において各成分を混合し、８０℃まで加温し徐々に冷却し油性軟膏を得た。

【００３５】
［試験例３］
実施例３で調製した油性軟膏を、試験例１に記載の抗酸化効果及び過酸化脂質生成抑制効果の評価試験法に示す方法に準じ各試験供した。その結果、該油性軟膏は（Ｂ）成分を配合しない以外は、全ての成分を含む油性軟膏と比べて、ＤＰＰＨラジカル消去作用及び生体膜脂質過酸化の抑制効果に優れていた。
【００３６】
【発明の効果】
本発明の抗酸化剤を含む化粧品、医薬品、医薬部外品、食品等の組成物は、生体内に生成した活性酸素や過酸化脂質によって引き起こされる障害を抑制する効果がある。従って、健康上、美容上の障害についての治療に有効であり、さらに飲食品の安定化・保存性の向上にも有用である。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to an antioxidant and a composition containing the antioxidant. Furthermore, the present invention relates to an external preparation for skin that can be used in the fields of pharmaceuticals, quasi-drugs, cosmetics and the like containing the antioxidant.
[0002]
[Prior art]
In recent years, it has become clear that active oxygen generated in a living body reacts with unsaturated fatty acids to generate lipid peroxide, which has an adverse effect on the human body. For example, active oxygen ischemic injury and radiation injury, lipid peroxide and its oxidative degradation products act on nucleic acids and proteins, and cause arteriosclerosis, hypertension, vascular injury, hepatic dysfunction, retinopathy caused by them. And cataracts. In particular, the skin is directly stimulated by environmental factors such as ultraviolet rays, so active oxygen is likely to be generated, increasing the active oxygen concentration, abnormal pigmentation such as spots and freckles such as lipid peroxide, inflammation, edema, The effects are remarkable such as necrosis, wrinkles and aging.
[0003]
In addition, many cosmetics, pharmaceuticals, foods and beverages contain fats and oils, which react with active oxygen during storage and use to generate lipid peroxides, resulting in reduced quality and reduced nutrition and the human body. The development of toxicity is a major problem.
[0004]
For this reason, in order to improve the abnormalities of lipid peroxide in the living body, research for searching for a drug having an antioxidant effect has been widely conducted. Representative examples include fat-soluble tocopherol (vitamin E) and water-soluble ascorbic acid (vitamin C) as natural product antioxidants, and BHT (3,5-tert-butyl-) as a synthetic antioxidant. Examples thereof include 4-hydroxytolen and BHA (2, (3) -tert-butyl-hydroxyanysol) and the like, but their effects were not satisfactory.
[0005]
On the other hand, many attempts have been made to obtain a substance having a high antioxidant activity in order to improve the abnormalities of lipid peroxide in the living body, and various crude drug extracts have been disclosed (for example, Japanese Patent Application Laid-Open No. Hei 5- 246877, JP-A-8-92053, JP-A-8-301745).
[0006]
[Problems to be solved by the invention]
However, the crude drug extract has a somewhat higher antioxidant activity than tocopherol, ascorbic acid, etc., but the activity is not satisfactory. Further, the synthetic antioxidants BHT and BHA have problems such as suspected carcinogenicity. Therefore, in addition to these antioxidants, there is a demand for a substance which has similar means for suppressing the production of lipid peroxide and is highly safe.
[0007]
[Means for Solving the Problems]
In view of such a situation, the present inventors have conducted intensive studies in an attempt to improve the problems of the prior art. (TILO)] was found to have a strong free radical scavenging action and a lipid peroxide production inhibitory action.
[0008]
That is, the present invention provides an antioxidant characterized by containing an extract of Tilia mexicana (TILO), and a composition such as a cosmetic, a food, or a medicine containing the antioxidant. Is provided.
[0009]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, embodiments of the present invention will be described in detail.
Tilia mexicana used in the present invention is a plant belonging to the genus Lindenaceae, native to Mexico.
[0010]
The extract of Tilia mexicana used in the present invention is obtained by extracting from a part or all of a plant such as leaves, stems, flowers, seeds, fruits, bark, rhizomes, roots and the like of the plant. It is something that can be done. Preferably, it is obtained by extracting from one of the leaves or the stem, or a mixture of both. In general, dried or raw plants are used as they are or cut. As the extraction solvent to be used, the following extraction solvent is used in an amount of 5 to 50 parts based on the dry matter of the dried or fresh plant.
[0011]
As the extraction solvent, generally, water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohol (1,3-butylene glycol) , Propylene glycol, etc.), lower alkyl esters (ethyl acetate, etc.), hydrocarbons (benzene, hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (diethyl ether, tetrahydrofuran, dipropyl ether), acetonitrile, etc. Is mentioned. 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, methanol, propylene glycol, etc.) is used.
[0012]
Although the extraction method is not particularly limited, the extraction is carried out at normal temperature or under heating, and examples of the method include ordinary extraction and Soxhlet extraction. The extraction time is not limited, but is generally preferably 1 hour to 1 week.
[0013]
The extract may be used as it is, or may be subjected to various treatments before use. For example, a concentrated solution obtained by concentrating them under normal pressure or reduced pressure, or a solid obtained by evaporating the solvent in the concentrated solution to dryness, or a solid obtained by crystallization from the concentrated solution and drying by filtration, or freezing the concentrated solution Dry solids and the like can be mentioned.
[0014]
The amount of the extract of Tilia mexicana (Tilia mexicana) according to the present invention per dry matter as an antioxidant is not particularly limited, but is 0.001 to 20.0% by weight based on the total amount. (Hereinafter referred to as “wt%”), particularly preferably 0.01 to 10 wt%.
[0015]
The composition containing the antioxidant of the present invention is characterized by blending the above-mentioned antioxidant, and its use is optional, but it is widely used in cosmetics, foods, pharmaceuticals, quasi-drugs, toiletries and the like. Can be.
[0016]
The cosmetics to which the present invention is applied are not particularly limited in dosage form, and include, for example, lotions, emulsions, creams, foundations, packs, lipsticks, facial cleansers, shampoos, rinses, hair tonics, and the like. These cosmetics include various components generally used in cosmetics, that is, aqueous components, oily components, powder components, alcohols, esters, surfactants, humectants, whitening components, ultraviolet absorbers, thickeners, Components such as coloring agents, fragrances, antioxidants, pH adjusters, chelating agents, preservatives and the like can be added.
[0017]
The food to which the present invention is applied is not particularly limited, and can be obtained, for example, by adding a required amount of an extract to each food raw material as a general food and processing and manufacturing the same by a normal manufacturing method.
[0018]
Pharmaceuticals to which the present invention is applied, quasi-drugs, the dosage form is not particularly limited, for example, tablets, granules, capsules, oral medicines such as drenches, ointments, cataplasms, creams, solutions and the like Examples include external preparations, injections such as sterile solutions and suspensions, and bath preparations. These pharmaceuticals can take the required unit dosage forms with physiologically acceptable vehicles, carriers, excipients, binders, stabilizers, flavoring agents, and the like. For example, compositions for tablets and capsules include binders such as tragacanth, acacia, gelatin, excipients such as microcrystalline cellulose, gelatinized starch, leavening agents such as alginic acid, lubricating agents such as magnesium stearate. A sweetener such as sucrose, lactose and saccharin, a flavoring agent such as peppermint, reddish oil and cherry can be mixed together and formulated by a usual method. Sterile compositions for injections also dissolve the active substance in vehicles such as water for injection, naturally occurring vegetable oils such as sesame oil, coconut oil, peanut oil, cottonseed oil, or synthetic fat vehicles such as ethyl oleate. Alternatively, it can be formulated by a usual method of suspending. As an external preparation, vaseline, paraffin, oils and fats, lanolin, macrogol and the like are used as bases, and ointments and creams are prepared by a usual method.
[0019]
The skin external preparation of the present invention means any dosage form that can be used externally, for example, lotion, emulsion, cream, foundation, pack, essence, lipstick, facial cleanser, gel, aerosol, ointment, poultice, Examples include pastes, plasters, baths, detergents and the like applied to the skin, and shampoos, rinses, treatments, hair tonics and other hairs. Further, the external preparation for skin of the present invention can be used for any of medicine, quasi-drug, and cosmetic.
[0020]
The skin external preparation of the present invention includes an aqueous component, an oily component, a powder component, a wax, fatty acids, alcohols, esters, a surfactant, a humectant, and a whitening component, which are components used in ordinary skin external preparations. A UV absorber, a thickener, a coloring agent, a fragrance, an antioxidant, a pH adjuster, a chelating agent, a preservative, and the like can be appropriately compounded within a range that achieves the object of the present invention.
[0021]
【Example】
Hereinafter, the present invention will be described in detail based on examples. Note that the present invention is not limited to these.
[0022]
[Extraction Example 1] (Production of Tilia mexicana extract I)
After 100 g of dried Tilia mexicana was extracted with 1 L of methanol at room temperature for 24 hours, and then extracted twice in the same manner, the combined extracts were concentrated and dried to obtain a powdery solid. 7 g were obtained.
[0023]
[Extraction Example 2] (Production of Tilia mexicana extract II)
100 g of dried Tilia mexicana was placed in 1 L of water / ethanol (1: 1, volume ratio) solvent, extracted while stirring at room temperature for 24 hours, filtered, and the filtrate was concentrated to dryness. 8.5 g of a powdery solid was obtained.
[0024]
[Extraction Example 3] (Production of Tilia mexicana extract III) 100 g of dried Tilia mexicana was mixed with 1 L of a mixed solution (1: 1) of 1,3-butylene glycol and water. After extraction at room temperature for 10 days, the mixture was filtered, and the filtrate was concentrated to dryness to obtain 9 g of a powdery solid.
[0025]
[Test Example 1]
The evaluation test methods used in the present invention for examining the antioxidant ability and the ability to inhibit lipid peroxide production of the above extract are as follows. For comparison, dl-α-tocopherol and catechin (manufactured by Nacalai Tesque, Inc.), which are already known to have antioxidant activity, were evaluated in the same manner, and the radicals at 100 ug / ml and 10 ug / ml of the sample were evaluated. The removal rate (%) and the lipid peroxidation inhibition rate (%) were determined.
(1) DPPH radical scavenging test The inhibitory effect of the above extract on the radical chain reaction of lipids was examined using an ethanol solution of the neutral radical Diphenyl-p-picrhydrhydril (DPPH). 800 ul of 250 mM acetate buffer (pH 5.5) is mixed with 800 ul of ethanol and 20 ul of a sample, and pre-incubated at 30 ° C. for 5 minutes. To this solution, 400 ul of 250 uM DPPH / ethanol solution was added and mixed, left at 30 ° C. for 30 minutes, and the absorbance at 517 nm was measured. Then, 20 ul of 300 ug / ml BHT (3,5-tert-butyl-4-hydroxytolen) was added. Then, the absorbance when the DPPH radical is completely removed is measured. From this difference, the radical removal rate is determined.
[0026]
(2) Lipid peroxide production inhibition effect test (lipid peroxidation in mitochondria)
In order to examine the ability of biological lipids to inhibit peroxidation, the ability of mitochondria to inhibit lipid peroxidation was measured with reference to the method of Takayanagi et al. (Takayanagi et al., Biochem. J. 192: 853-860, 1980). 50 ul of a mitochondrial fraction and 10 ul of a sample prepared as usual from liver isolated from a rat (Wistar male, body weight 100-150 g) were added to the following solution.

After keeping the above mixture at 37 ° C. for 5 minutes, 100 ul of 1 mM NADH was added and reacted at 37 ° C. for 15 minutes. 90 ul, 2% BHT and 2 ml TCA-TBA-HCl solution were added and mixed, followed by heating at 100 ° C. for 15 minutes. After cooling to room temperature, the mixture was centrifuged at 5000 rpm for 10 minutes, and the absorbance (535 nm) in the supernatant was measured.
The results are shown in Table 1 (Table 1). The Tilia mexicana extracts of Extraction Examples 1, 2, and 3 have a DPPH radical scavenging action comparable to dl-α-tocopherol and catechin, which are already known to have antioxidant activity, and a lipid peroxidation in mitochondria. It was found to have an effect of suppressing oxidation.
[0027]
[Table 1]

[0028]
[Example 1] (cream)
The B component was mixed with the A component in the following blending amount, and the mixture was uniformly heated and melted to a temperature of 80 ° C. Next, the C component was injected, stirred and mixed, and then cooled to 30 ° C. with stirring to obtain a cream.

[0029]
[Comparative Example 1] (cream)
All were prepared in the same manner as in Example 1 except that the component (B) Tilia mexicana extract was omitted, and used in each of the tests described above.
[0030]
[Example 2] (two-phase lotion)
The component A was uniformly mixed and dissolved at room temperature in the following amount, and the component B was slowly added by stirring to obtain a two-phase lotion.

[0031]
[Comparative Example 2] (Two-phase lotion)
All were prepared in the same manner as in Example 2 except that the component (B) Tilia mexicana extract was omitted, and used in each of the tests described above.
[0032]
[Test Example 2]
Examples 1 and 2 The effects of the compositions obtained in Comparative Examples 1 and 2 were evaluated by the following test methods of a usefulness evaluation test.
(1) Utility evaluation test Eighty healthy women (25 to 40 years old) were divided into four groups of 20 each, and the samples of Examples 1 and 2 and Comparative Examples 1 and 2 were applied twice a day, respectively. A questionnaire survey was conducted to evaluate the effect of preventing aging (prevention of rough skin, gloss and firmness of skin) three months after the start of application. The evaluation criteria of the questionnaire were evaluated as "excellent" for valid items, "good" for slightly effective items, "acceptable" for slightly effective items, and "impossible" for invalid items. went.
As shown in Table 2 (Table 2), better results were obtained in Examples 1 and 2 than in Comparative Examples 1 and 2.
[0033]
[Table 2]

[0034]
[Example 3] (oil-based ointment)
The components were mixed in the following amounts, heated to 80 ° C., and gradually cooled to obtain an oily ointment.

[0035]
[Test Example 3]
The oily ointment prepared in Example 3 was subjected to each test according to the method shown in Test Method for Evaluation of Antioxidant Effect and Lipid Peroxide Production Inhibiting Effect described in Test Example 1. As a result, the oil-based ointment was superior to the oil-based ointment containing all the components except that the component (B) was not added, in the DPPH radical scavenging action and the effect of suppressing the lipid peroxidation of biological membranes.
[0036]
【The invention's effect】
Compositions such as cosmetics, pharmaceuticals, quasi-drugs, and foods containing the antioxidant of the present invention have an effect of suppressing disorders caused by active oxygen and lipid peroxide generated in a living body. Therefore, it is effective for treatment of health and cosmetic disorders, and is also useful for stabilizing food and drink and improving storage stability.

Claims (3)

An antioxidant comprising an extract of Tilia mexicana. A composition comprising the antioxidant according to claim 1. An external preparation for skin, comprising the antioxidant according to claim 1.