JP3929369B2 - Cosmetics - Google Patents

Description

【００２９】
結果は表３のようになった。

【００３０】
使用テスト２
使用テスト１と同様な試験を行った。但し、比較例は実施例より製造例
を同等の牛由来のコラーゲンに置き換えてテストを行った。
【００３１】
実験の方法は表４のように行った。

【００３２】
結果は表５のようになった。

【００３３】
使用テスト３（官能評価）
製造例１と製造例１と同濃度の牛由来コラーゲン水溶液を用いて官能評価を行った。選抜した官能パネラー３名に前腕内側部で評価を行ったところ、以下の評価を得た。なお、評価基準は製造例１の方が非常によいが５、製造例１と比較例と同程度が３、比較例の方が非常によいが１として、その中間は０．５刻みで評価点をつけてもらった。３人の平均点を以下の表６に示す。
【００３４】

【００３５】
以上のように本発明の１型（α１）₃ホモ分子構造をもつコラーゲン、およびその配合した実施例の有効性が高いことがわかった。製造例が１型(α1)₃ホモ分子コラーゲンの構造であることを確かめるために、製造例１をＳＤＳポリアクリルアミドゲル電気泳動を行った。結果を図１に示す。
【００３６】
【図１】

【００３７】
図のＡは分子量マーカーを、Ｂは牛由来アテロコラーゲンを、Ｃは牛由来コラーゲンを、Ｄは製造例１を示す。このようにＢとＣはコラーゲンを構成する３つの鎖が大きく分けて２種類の分子量のものから構成されているが、製造例１は同一のほぼ同一の３つの鎖から構成されており、１型(α1)₃ホモ分子構造をとるコラーゲンであることがわかる。
【００３８】
【効果】
１型(α1)₃ホモ分子コラーゲン（貝類肉より得た）を配合することを特徴とする皮膚外用剤はシワの減少、肌のつや、肌のはりを改善し、その結果、貝類肉が廃棄物のして海洋汚染を引き起こすことを防ぐ。また、１型(α1)₃ホモ分子コラーゲンを加水分解、アテロ化、アシル化などの処理を行うことによって安全性が増し、溶解性が改善され、よリ多くの用途に用いることができた。[0001]
[Industrial application fields]
The present invention relates to a cosmetic characterized in that it contains collagen that is useful for the skin.
[0002]
[Prior art]
Collagen is a major protein that forms the skin of animals such as cow skin, pig skin, and fish skin. Collagen is an important connective tissue protein that accounts for 70% of the dermis and is involved in the formation of skin firmness and wrinkles, and has high moisturizing properties and affects the freshness of the skin. Most of the collagen present in the skin is type 1 and the rest is type 3. However, it is damaged by aging and sunlight. In order to prevent this, it has been used in large quantities as a raw material for cosmetics. They are also used in pharmaceuticals and medical devices because of their excellent biocompatibility. In recent years, it has also attracted attention as a food material. Collagens that have been used so far have been mainly derived from cow skin and pork skin, but in recent years they are also affected by viruses, etc., and the structure of collagen differs depending on the origin, and collagens of various origins have been developed. ing.
However, currently used cattle, pigs, fish skins, etc. are all collagens having a type 1 (α1) ₂ α2 heteromolecular structure.
In addition, even for shellfish meat, Akoya oysters used for scallop and pearl farming have been treated as waste, except for scallops. Already, the patents used in the patent registration Nos. 1835341, 1805187, and 1929815 have been disclosed, but at present, many of them are discarded as waste.
[0003]
[Problems to be solved by the invention]
In view of the current situation as described above, the present inventors have developed a highly effective and highly safe collagen and attempted to apply it to pharmaceuticals, quasi drugs, cosmetics and foods.
[0004]
[Means for Solving the Problems]
Collagen is already known to be very useful for the skin as described above, but collagen derived from mammals is not desired by some people and responds to various needs Therefore, as a result of intensive studies, it was found that collagen having a type 1 (α1) ₃ homomolecular structure obtained from shellfish meat meets the gist of the present invention. Collagen with type 1 (α1) ₃ homo-molecular structure can be obtained from chicken embryo tendon, but the most economically advantageous is from shellfish meat, and oysters, abalone, clams, etc. are shellfish Is used as food, but in the case of pearl oysters, it is currently dumped as waste at the end of pearl cultivation, and the most effective use of pearl oysters is as a raw material of the present invention.
Collagen with this type 1 (α1) ₃ homomolecular structure is one type often found in the skin, and type 1 (α1) ₂ α2 obtained from cattle and bonito skins that have been used so far as described below. Moisture retention and the like are better than collagen having a heteromolecular structure, and the mechanism of action of this result is not clearly understood, but it seems to be caused by the difference in structure.
[0005]
The pearl oysters have been cultured for pearls, and when the pearls are removed, the scallops collect the shellfish after removing the scallops. Water is added to this and stirred to collect insoluble matter. In the case of other shellfish, use shellfish without shell pillars if necessary. The method for extracting collagen may be performed by a known method. That is, for example, after adding water, after stirring, a method of separating with a centrifuge or a method of leaving the supernatant after standing still may be selected. In order to further remove impurities from this insoluble matter, salt solutions such as sodium chloride, potassium chloride, sodium acetate, sodium citrate, sodium hydrogen phosphate, sodium hydroxide, potassium hydroxide, calcium hydroxide, aqueous ammonia, etc. A basic solution or a chelating agent such as ethylenediaminetetraacetate is added and stirred to collect insoluble matter. Further, after removing impurities with a water-soluble organic solvent such as ethanol, collagen is obtained by acid extraction using acetic acid, citric acid, lactic acid, hydrochloric acid, phosphoric acid or the like. The concentration varies depending on the type of acid and various conditions, but the concentration is 0.01 to 2 mol. When this liquid is added and stirred for 2 to 48 hours, it is extracted. In addition, temperature is 30 degrees C or less, Preferably it is 2-10 degreeC.
In addition, after using an enzyme, it may be extracted by a known method such as a method of extracting with an acid. Further, depending on the application, it may be better to further purify, and it is purified using a hydrophilic organic solvent or salting out method.
Type 1 (α1) ₃ Collagen or protein other than collagen having a homomolecular structure is included in some cases, but there is no problem in actual use. It is possible to change the degree of purification from the economical aspect and safety aspect depending on the application.
[0006]
The collagen obtained as described above may be used as it is, but in order to further expand the range of use, treatments such as hydrolysis, atelolation and acylation are added depending on the use.
The hydrolysis method includes proteases such as collagenase, papain, and pepsin, inorganic acids such as hydrochloric acid, sulfuric acid, and nitric acid, organic acids such as acetic acid, lactic acid, and succinic acid. Remove unnecessary enzymes and acids as needed. Moreover, there is no problem even if a decomposition method such as an alkali other than an enzyme or an acid is used.
When using only an enzyme, collagen other than collagenase is not decomposed, so it is used in combination with collagenase or after being decomposed with an acid.
[0007]
Atelolation is the removal of telopeptides present at the ends of the helical structure of collagen. Since these telopeptides have antigen production, they may be required for some applications. In this method, by using a proteolytic enzyme other than collagenase, the helical structure portion which is the main body of collagen is not decomposed, and only telopeptide can be removed.
[0008]
In the acylation, part or all of the side chain amino group of collagen is acylated using a saturated or unsaturated fatty acid having 2 to 22 carbon atoms or a dicarboxylic acid having 2 to 8 carbon atoms. Acylation improves the solubility and emulsification properties, depending on the fatty acid or dicarboxylic acid used.
In addition, there are a method of using a plurality of fatty acids or dicarboxylic acids when acylating, a method of partially acylating and then acylating again with a fatty acid or dicarboxylic acid different from the fatty acid or dicarboxylic acid used first.
Examples of fatty acids or dicarboxylic acids used for acylation include acetic acid, butyric acid, propionic acid, caproic acid, caprylic acid, capric acid, heptanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, ricinoleic acid, oleic acid , 12-hydroxystearic acid, arachidic acid, behenic acid, malonic acid, succinic acid, glutaric acid, maleic acid, 2-methylmaleic acid, 2,3-dimethylmaleic acid, phthalic acid, but of course other fatty acids Of course, dicarboxylic acids can also be used.
In particular, the case of using succinic acid is called succinylation, and this treatment is particularly effective.
In the acylation method, the pH of collagen is adjusted to 8 to 14, and a necessary amount is obtained by adding a necessary amount of a fatty acid or dicarboxylic acid anhydride while maintaining a low temperature (25 ° C. or less).
The above-mentioned hydrolysis, atelolation and acylation are combined as necessary to obtain the necessary collagen.
[0009]
This is blended into pharmaceuticals, quasi drugs, cosmetics and foods. The collagen or collagen derivative of the present invention can be used without particular limitation as long as it is a field or product in which collagen or a collagen derivative has already been used.
Since the dosage form is not particularly limited, in the case of pharmaceuticals, quasi drugs, cosmetics, creams, ointments, packs, gels, tablets, powders, solutions, tablets, powders, granules, dragees, capsules, suspensions, Select any dosage form such as ampoules, injections, skin cosmetics such as lotion, milky lotion, cream, makeup base lotion, makeup base cream, liquid or cream or ointment type foundation, eye color, Makeup cosmetics such as teak color, shampoos, rinses, hair treatments, permanent liquids, hair conditioners, hair dyes, hair nicks, hair cosmetics such as hair growth and hair nourishing bodies, hand creams, leg creams, body lotions, etc. Used for cosmetics.
In the case of food, beverages such as soup, fruit juice, vegetable juice, whey drink, soft drink, ice cream, gum, candy, rice cake, pastry, Japanese confectionery, jam, marmalade, cheese, butter, yogurt, etc. It can be blended into general foods and drinks such as tablets and capsule-shaped health foods.
Moreover, although the mixing | blending ratio is arbitrary, it mix | blends in 0.01 to 100% of range.
[0010]
For pharmaceuticals, quasi-drugs, and cosmetics, natural animal and vegetable oils and fats such as olive oil, mink oil, castor oil, palm oil, beef tallow, evening primrose oil, coconut oil, castor oil, cacao are used to formulate these dosage forms. Oil, macadamia nut oil, etc .; wax such as jojoba oil, beeswax, lanolin, carnauba wax, candelilla wax, etc .; higher alcohols such as lauryl alcohol, stearyl alcohol, cetyl alcohol, oleyl alcohol; higher fatty acids such as lauric acid, palmitic acid, Stearic acid, oleic acid, behenic acid, lanolin fatty acid, etc .; higher aliphatic hydrocarbons such as liquid paraffin, solid paraffin, squalane, petrolatum, ceresin, microcrystalline wax, etc .; synthetic ester oils such as butyl stearate, hexyl laurate, Diiso Lopyl adipate, diisopropyl sebacate, octyldodecyl myristate, isopropyl myristate, isopropyl palmitate isopropyl myristate, cetyl isooctanoate, neopentyl glycol dicaprate; silicone derivatives such as silicone oils such as methyl silicone and methyl phenyl silicone [0011]
Surfactants include anionic surfactants such as alkyl sulfates, fatty acid salts, alkyl phosphates, polyoxyethylene alkyl ether phosphates and sulfates, etc .; nonionic surfactants such as glycerin fatty acid esters , Polyoxyethylene sorbitan fatty acid ester, polyoxyethylene alkyl ether, sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyglycerin fatty acid ester, etc .; double-sided activators such as alkylbetaines, phosphobetaines, phosphatidylcholines, phosphatidylethanolamines, phosphatidylserine , Phosphatidylglycerol, phosphatidylinositol, and their lysates, as well as phosphophatidic acid and its salts
Polyhydric alcohols such as ethylene glycol, diethylene glycol, triethylene glycol, higher polyethylene glycols, propylene glycol, dipropylene glycol, higher polypropylene glycols, 1,3-butylene glycol, 1,4-butylene glycol, etc. Butylene glycols, glycerol, diglycerol, higher polyglycerols, sugar alcohols such as sorbitol, mannitol, xylitol, maltitol, ethylene oxide of glycerol (hereinafter abbreviated as EO), propylene oxide (hereinafter abbreviated as PO) ) Adduct, EO of sugar alcohols, PO adduct, monosaccharides such as galactose, glucose, fructose and its EO, PO adduct, polysaccharide such as maltose, lactose and its EO, P Polyhydric alcohol such as adduct [0013]
Tocopherol, tocopherol acetate, vitamin C, allantoin, placental extract, elastin, arbutin, collagen, triclosan, trichlorocarban, dipotassium glycyrrhizinate, methylparaben, butylparaben as drugs
These materials are combined to obtain the necessary preparation. However, the surfactant has one advantage even if it is not blended as described in JP-A-3-83909, JP-A-5-43416, and JP-A-6-9333, and it is not blended depending on the purpose. Of course, other raw materials are blended as necessary.
Similarly, in the case of food, a desired food or drink is prepared using arbitrary raw materials.
[0015]
Although the manufacture example, Example, etc. which are the actual utilization methods of this invention are described below, this invention is not limited to these manufacture examples and Examples at all.
[0016]
Production Example 1 (Akoya shellfish type 1 (α1) ₃ homomolecular collagen)
After the pearl culture is finished, Akoya shellfish collects the shellfish after the pearl is taken out. 880 g of the obtained Akoya shellfish is salted with salt to remove the slime, then homogenized in 1200 mL of 0.6 M KCl for 2 hours, and the shellfish is collected by decantation. Then, after stirring in 1200 mL of 0.45 M NaCl for 2 hours, shellfish was collected by decantation to obtain 146 g of fragmented pearl oyster. Furthermore, this piece shellfish is dissolved in 2N sodium hydroxide solution, neutralized to pH 6-7, the supernatant is applied to a dialysis membrane (Wako Pure Chemicals: Dialysis Membrane 36), and the external solution is purified water. Dialysis was performed overnight.
[0017]
Production Example 2 (Clamshell type 1 (α1) ₃ homomolecular collagen)
The same operation as in Production Example 1 was performed using peeled clams.
[0018]
Production Example 3 (Akoya shellfish type 1 (α1) ₃ homomolecular atelocollagen)
In the process of preparing the purified collagen of Production Example 1, 14.6 g of a piece of pearl oyster meat obtained after treatment in 1200 mL of 0.45 M NaCl was stirred overnight in 1000 mL of 0.1 M acetic acid, and then centrifuged (3000 × g, 20 min) and collect the precipitate (68 g). The obtained precipitate was dispersed in 1200 mL of 0.01N HCl, added with 1.0 to 1.5 g of pepsin, stirred overnight, centrifuged (9500 × g, 2 hours), and the supernatant was dialyzed. (Wako Pure Chemical: Dialysis membrane 36) was dialyzed overnight using purified water as the external solution.
[0019]
Production Example 4 (Akoya shellfish type 1 (α1) ₃ homomolecular succinylated collagen)
To 10 g obtained in Production Example 1, 500 ml of purified water was added, and 0.5 g of succinic anhydride was gradually added while maintaining the pH at 10 ° C. and NaOH. This was lyophilized.
[0020]
Production Example 5 (Akoya shellfish type 1 (α1) ₃ homomolecular atelocollagen hydrolyzate)
1 g of the freeze-dried product of Production Example 3 was added to 10 mL of 6N HCl, heated at 100 ° C. for 2 hours, cooled, and then freeze-dried. The crude reaction product was hydrolyzed to type 1 (α1) 3 homomolecular collagen. It is a thing.
[0021]
Example-1 Lotion (parts by weight)
Olive oil 0.5
Polyoxyethylene (20E.0) sorbitan monostearate 2.0
Polyoxyethylene (60E.0) hydrogenated castor oil 2.0
Ethanol 10.0
1.0% sodium hyaluronate aqueous solution 5.0
Production Example 1 5.0
Purified water 75.5
Prepared according to conventional methods.
[0022]
Example-2 Cream (parts by weight)
Squalane 20.0
Olive oil 2.0
Mink oil 1.0
Jojoba oil 5.0
Beeswax 5.0
Cetostearyl alcohol 2.0
Glycerol monostearate 1.0
Sorbitan monostearate 2.0
Purified water 47.9
Production Example 2 0.5
Polyoxyethylene (20E.0) sorbitan monostearate 2.0
Polyoxyethylene (60E.0) hydrogenated castor oil 1.0
Glycerin 5.0
1.0% sodium hyaluronate aqueous solution 5.0
Methyl paraoxybenzoate 0.1
Prepared according to conventional methods.
[0023]
Example-3 hair shampoo (parts by weight)
N-coconut oil fatty acid acyl-L-glutamic acid triethanolamine 30% aqueous solution 10.0
Lauric acid diethanolamide 5.0
Lauroylmethyl taurine sodium 6.0
1,3 Butylene glycol 3.0
Phenoxyethanol 1.0
Methyl paraoxybenzoate 0.5
[2-hydroxy-3-trimethylammoniopropyl] chloride
Hydroxyethyl cellulose 0.2
Production Example 3 2.0
Purified water 72.3
Prepared according to conventional methods.
[0024]
Example-4 Hair growth lotion (parts by weight)
Ethanol 20.0
Methyl paraoxybenzoate 0.1
Hinokitiol 0.01
Acetic acid dl-α-tocophenol 0.1
Polyoxyethylene (60E.0) hydrogenated castor oil 0.5
Purified water 71.79
Production Example 4 1.0
Assembly extract 3.0
Carrot extract 3.0
Pantothenyl ethyl ether 0.5
Prepared according to conventional methods.
[0025]
Example-5 Tablet (parts by weight)
Production Example 5 40.0
Lactose 20.0
Cornstarch 16.0
Crystalline cellulose 20.0
Polyvinylpyrrolidone 3.0
Mg stearate 1.0
According to a conventional method, each of the above components was granulated and compression-molded to prepare 1000 tablets of 500 mg tablets.
[0026]
Example-6 Beverage Production Example 1 1g
Xylitol 10g
Vitamin B1 hydrochloride 0.5mg
Vitamin B2 0.2mg
Vitamin C 500mg
Niacin 1.0mg
Pantothenic acid Ca 1.0 mg
100 g of purified water
[0027]
In order to confirm the above effectiveness, the following usage test was conducted.
Usage test 1
The faces of 16 females were divided into left and right, and one was used as an example and the other was used as a comparative example at least once daily, and a questionnaire was conducted one year later. In the comparative example, the production example was changed to water from the example. (Comparative Examples 1 and 2)
The criteria are as follows, and the results of the questionnaire are summarized in the following table.
The example is much better 3
The example is much better 2
Example is slightly better 1
No difference 0
Comparative example is slightly better -1
The comparative example is much better -2
The comparative example is much better -3
[0028]
The experimental method was as shown in Table 2.

[0029]
The results are shown in Table 3.

[0030]
Usage test 2
A test similar to the use test 1 was performed. However, the comparative example was tested by replacing the production example with an equivalent bovine collagen from the example.
[0031]
The experimental method was as shown in Table 4.

[0032]
The results are shown in Table 5.

[0033]
Usage test 3 (Sensory evaluation)
Sensory evaluation was performed using cattle-derived collagen aqueous solution having the same concentration as in Production Example 1 and Production Example 1. When the selected sensory panelists were evaluated on the inner side of the forearm, the following evaluations were obtained. In addition, although the evaluation standard is much better in the production example 1, 5 is the same as the production example 1 and the comparative example, and the comparative example is much better, but the middle is evaluated in 0.5 increments. I was given a point. The average score for the three is shown in Table 6 below.
[0034]

[0035]
As described above, it was found that the collagen having the type 1 (α1) ₃ homomolecular structure of the present invention and the examples in which the collagen was blended were highly effective. In order to confirm that the production example has the structure of type 1 (α1) ₃ homomolecular collagen, the production example 1 was subjected to SDS polyacrylamide gel electrophoresis. The results are shown in FIG.
[0036]
[Figure 1]

[0037]
In the figure, A represents a molecular weight marker, B represents bovine-derived atelocollagen, C represents bovine-derived collagen, and D represents Production Example 1. Thus, B and C are composed of two types of molecular weights roughly divided into three chains constituting collagen, but Production Example 1 is composed of three identical and almost identical chains. It can be seen that the collagen has a type (α1) ₃ homomolecular structure.
[0038]
【effect】
An external preparation for skin characterized by blending type 1 (α1) ₃ homomolecular collagen (obtained from shellfish meat) reduces wrinkles, improves skin shine and skin elasticity, and as a result, shellfish meat is discarded. Prevent marine pollution caused by things. In addition, the type 1 (α1) ₃ homomolecular collagen was subjected to treatments such as hydrolysis, atelolation, and acylation, thereby increasing safety and improving solubility, so that it could be used in many applications.

Claims (4)

A cosmetic product obtained by extracting shellfish meat and blending collagen having a single band in molecular weight measurement by electrophoresis . The cosmetic according to claim 1, wherein the collagen is blended with one or more of hydrolysis, atelolation and acylation.   The cosmetic according to claim 2, wherein the hydrolysis is hydrolysis with acid or alkali.   The cosmetic product according to claim 1, wherein the shellfish meat is Akoya shellfish.