JPH0827192A - Method for producing modified collagen and cosmetic base agent - Google Patents

Abstract

PURPOSE:To obtain a modified collagen soluble in a pH range of <= a specific value, excellent in compounding stability and transparency, and useful as a cosmetic base agent for smoothening skin and hair, etc., by modifying an animal tissue containing collagen and subsequently extracting the modified product. CONSTITUTION:The method for producing the modified collagen useful as a cosmetic base agent for smoothening skin and hair, etc., comprises making paper from a pig skin immersed in a lime solution as a collagen-containing animal tissue, subjecting the paper to a decalcification treatment, a mincing treatment and a defatting treatment, esterifying the product in the presence of an alcohol such as a compound of formula I (R<1> is 1-20C alkyl, alkenyl), formula II (R<2> is H, 1-20C alkyl, alkenyl; n is an integer of 1-200), formula III (R<3> is H, 1-20C alkyl, alkenyl; n' is an integer pf 1-200) or formula IV (R<4> is H, OH, 1-20C alkyl, alkenyl, alkoxy; Ph is phenylene; n'' is an integer of 0-200) or glycerol and sulfuric acid at 27 deg.C for 7 days under shaking, filtering the solution, washing the filtrate, treating the washed filtrate with an enzyme in a 2% citric acid, and subsequently extracting the treated solution.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing modified collagen and a cosmetic base comprising the modified collagen derivative. More specifically, it relates to a cosmetic base that is incorporated into skin or hair cosmetics to moisturize and smooth the skin.

[0002]

BACKGROUND OF THE INVENTION Collagen is a major protein that forms connective tissue such as skin. Conventionally, acid-soluble collagen and atelocollagen have been used as cosmetic ingredients in lotions, emulsions, creams, etc., but their isoelectric point is pH 7.
Since it exists in the range of up to 9.5, it has poor solubility at pH 5 or higher and precipitates and aggregates. For this reason, it is difficult to use in the pH range of weak acidity to weak alkali that is often used in cosmetics, and it can be said that its formulation is limited. In addition, it has poor compatibility with high molecular weight alcohols such as polyethylene glycol used as a cosmetic compounding agent, and further forms a complex with thickeners such as hydroxyethyl cellulose and carboxymethyl cellulose and precipitates. Therefore, the compounding is impossible.

In order to overcome the above problems, for example, Japanese Patent Application No.
3-102178 proposes succinylated collagen, methyl esterified collagen, ethyl esterified collagen, and Japanese Patent Application No. 63-215770 proposes modified collagen such as acylated collagen.

However, the conventional method is to extract collagen from animal skin and then modify it. Extraction of collagen is performed by salting out after enzymatic reaction in an aqueous solution. On the other hand, the esterification reaction must be carried out in an organic solvent under anhydrous conditions. Therefore, in order to esterify collagen extracted in an aqueous solution,
The collagen obtained by extraction and salting out is desalted by an operation such as dialysis, and then dried by freeze-drying or the like. It became complicated and led to an increase in cost. In addition, biological materials such as collagen,
It is known that the resistance to external conditions such as heat is excellent when the higher order structure is maintained in the living body, but the resistance is remarkably reduced when separated into molecular units. Collagen, particularly that extracted as atelocollagen, undergoes thermal denaturation at a relatively low temperature, and therefore, when the reaction is carried out under heating conditions, it must react at a temperature lower than this. For this reason, the reaction rate is reduced, which causes an increase in manufacturing cost.

[0005]

The present invention has been made to overcome such problems, and is a method for inexpensively producing a collagen derivative which is soluble at pH 9 or less and has excellent compounding stability and transparency. And to provide a cosmetic base that smoothes the skin and hair.

[0006]

[Means for Solving the Problems] In order to solve the above problems, as a result of intensive investigations by the present inventors, as a result of performing a modification reaction of collagen in the state of the tissue of the animal, and then performing an extraction operation, The present invention has been completed and the present invention has been completed.

That is, the present invention relates to a method for producing a modified collagen, which comprises extracting the modified collagen after modifying the animal tissue containing the collagen.

The animal tissue containing collagen in the present invention is a tissue such as skin, tendon or cartilage of animals such as pigs, cows and sharks. These animal tissues may be subjected to physical treatments such as mincing, as well as treatments such as decalcification and degreasing that do not damage the higher-order structure of collagen.

In the present invention, the modification means to esterify a carboxy group existing in a collagen molecule with an alcohol, or to acylate an amino group existing in a collagen molecule with an acid chloride or an acid anhydride. Is.

In the present invention, examples of the alcohol used for esterification include those represented by the general formula (1)

[0011]

Embedded image

(Wherein R ¹ represents a linear or branched alkyl or alkenyl group having 1 to 20 carbon atoms), the general formula (2)

[0013]

[Chemical 6]

(Wherein R ² is H, a linear or branched alkyl or alkenyl group having 1 to 20 carbon atoms, and n is 1 to 1).
Represents an integer of 200),

[0015]

[Chemical 7]

(Wherein R ³ is H, a linear or branched alkyl group having 1 to 20 carbon atoms or an alkenyl group, and n ′ is 1
Represents an integer of ~ 200),

[0017]

Embedded image

(In the formula, R ⁴ is H, a hydroxyl group or a carbon number of 1
To 20 linear or branched alkyl groups, alkenyl groups, alkoxyl groups, Ph is a phenylene group, and n ″ is 0 to
It represents an integer of 200) and glycerin.

As the alcohol represented by the general formula (1), for example, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, dl-2-
Butanol, tert-butanol, 1-pentanol, dl-2-pentanol, 3-pentanol, 1
-Hexanol, dl-2- hexanol, dl-3
-Hexanol, 1-heptanol, 2-heptanol, 3-heptanol, 1-octanol, dl-2-
Octanol, 3-octanol, 1-nonanol,
2-nonanol, 3-nonanol, 4-nonanol, 5
-Nonanol, decanol, 1-undecanol, 2-
Undecanol and the like.

As the alcohol represented by the general formula (2) or (3), for example, ethylene glycol, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, ethylene glycol monophenyl ether, diethylene glycol, triethylene glycol. , Tetraethylene glycol, polyethylene glycol (M.
W. 200 to 20000), polyethylene glycol monomethyl ether (MW 350 to 5000), propylene glycol, 1,3-propanediol, propylene glycol monomethyl ether, dipropylene glycol, glycerin, 1,2-butanediol, 1,3
-Butanediol, 1,4-butanediol, 2,3-
Butanediol, 1,5-pentanediol, 2,4-
Pentanediol, 1,6-hexanediol, 2,5
-Hexanediol and the like.

Examples of the alcohol represented by the general formula (4) include hydroquinone, resorcin, pyrocatechol, phenol and salicyl alcohol.

The alcohols used in the present invention are not limited to those listed above, but considering the availability and productivity as cosmetic raw materials, ethylene glycol, propylene glycol, polyethylene glycol, hydroquinone. Are preferred.

The method for producing modified collagen of the present invention comprises the steps of performing a collagen modification reaction in the state of animal tissue,
The method is not particularly limited as long as it is a method of performing a step of performing an extraction operation, but for example, the following is performed.

First, after rawhide or lime-pickled skin is strained, demineralized (in the case of lime-pickled), minced and defatted animal tissue (pork skin, cowhide, etc.) is acid-catalyzed (hydrochloric acid, sulfuric acid,
p-toluenesulfonic acid or the like) or an enzyme (lipase, esterase or the like) and a solution of alcohol, glycol, phenol or the like, followed by ordinary esterification, and then washing the animal tissue containing the modified collagen,
Animal tissue containing modified collagen and protease (1/10 of substrate) in 0.5 M acetic acid solution.
Modified collagen is extracted by reacting 0) at 25 ° C. or lower, salting out the extract, re-dissolving after centrifugation, and modified collagen is obtained.

Esterification can also be performed by inducing a carboxyl group in a collagen molecule into a carboxylic acid chloride and dehydrochlorinating a hydroxyl group of alcohol, glycol or phenol with an acid chloride.

The modified collagen production method of the present invention does not require a desalting step such as dialysis and a dehydration step such as freeze-drying as in the conventional method. Therefore, the production cost is lower than that in the conventional method. It can be greatly reduced. The modified collagen obtained by the method of the present invention has the same performance as the modified collagen obtained by the conventional method.

The second invention of the present invention relates to a cosmetic base comprising the modified collagen produced by the above method.

The cosmetic base of the present invention can be applied to cosmetics in all applicable forms, such as lotions, lotions, cleansing foams, emulsions, packs, powders, foundations, shampoos, rinses, hair conditioners, etc. It is not limited to.

The base of the present invention is used in an amount of 0.01 to
By adding 0.5% by weight, the skin can be moisturized and the skin can be made smooth. 0.01
If it is less than 0.5% by weight, the moisturizing effect is not sufficient and
If the content exceeds the above range, a sticky feeling may occur, which is not preferable.

[0030]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1 10 g of demineralized, minced and degreased (hereinafter abbreviated as defatted pig skin) strained lime-pickled pig skin, 2 g of ethylene glycol, 2.86 ml of sulfuric acid (equivalent to 0.1 N) were added to a triangle. The mixture was placed in a flask and shaken at 24 ° C for 7 days. After shaking,
The defatted pig skin was filtered, added with distilled water, neutralized with a 0.1 N sodium hydroxide solution, and washed several times with 500 ml of distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product.

The results of subjecting the obtained product to an electrophoresis test are shown in FIG. From the results of FIG. 1, a shift of the band was confirmed as compared with the band of unmodified collagen, which shows that the collagen is esterified collagen. Further, since the same band as that of unmodified collagen does not exist, it can be seen that the extracted collagen is all modified collagen. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 2 10 g of defatted pig skin, 100 g of polyethylene glycol monomethyl ether (MW8000), acetonitrile 1
L and 2.86 ml of sulfuric acid (corresponding to 0.1 N) were charged in an Erlenmeyer flask, and shaken at 24 ° C. for 7 days. After shaking, the defatted pig skin was filtered, distilled water was added, neutralized with a 0.1 N sodium hydroxide solution, and washed several times with 500 ml of distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The obtained product was confirmed by the same method as in Example 1. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 3 Defatted pig skin 10 g, methyl alcohol 2 L, lipase 0.
Charge 2 g (2% of substrate) into an Erlenmeyer flask and
Shaking was performed at 7 ° C for 7 days. After shaking, the defatted pig skin was filtered and washed several times with 500 ml of distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product.
The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 4 Defatted pig skin 10 g, methanol 1 L, sulfuric acid 2.86 ml
(Equivalent to 0.1N) was charged in an Erlenmeyer flask and kept at 24 ° C for 7
Shaking was performed for a day. After shaking, the methanol was removed and n-
100 ml of dodecanol, 300 ml of dichloroethane,
A transesterification reaction was carried out by charging 2.86 ml of sulfuric acid and modified defatted pig skin into a 1-liter four-necked flask and refluxing under reduced pressure (60 ° C.) for 8 hours. Then, the defatted pig skin was filtered and washed with 500 ml of acetone, then distilled water was added to neutralize it with a 0.1 N sodium hydroxide solution and washed several times with 500 ml of distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 5 10 g of defatted pig skin, 1 L of propylene glycol, sulfuric acid 2.
Charge an Erlenmeyer flask with 86 ml (equivalent to 0.1 N), and
Shaking was performed at 4 ° C. for 7 days. After shaking, the defatted pig skin was filtered, distilled water was added, neutralized with a 0.1 N sodium hydroxide solution, and washed several times with 500 ml of distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 6 10 g of defatted pig skin, 100 g of polypropylene glycol monomethyl ether (MW6000), 1 L of acetonitrile, 2.86 ml of sulfuric acid (equivalent to 0.1N) were charged in an Erlenmeyer flask and shaken at 24 ° C. for 7 days. . After shaking, the defatted pig skin was filtered, washed several times with 500 ml of acetonitrile, and then distilled water was added to neutralize with 0.1 N sodium hydroxide solution and washed several times with distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 7 10 g of defatted pig skin, 11 g of hydroquinone (corresponding to 0.1 mol), 1 L of isopropyl ether, 2.86 ml of sulfuric acid
(Equivalent to 0.1N) was charged in an Erlenmeyer flask and kept at 24 ° C for 7
Shaking was performed for a day. After shaking, the defatted pig skin was filtered and washed several times with 500 ml of isopropyl ether. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 8 10 g of defatted pig skin, 100 g of polyoxyethylene nonylphenyl ether (MW9000), acetonitrile 1
L, 2.86 ml of sulfuric acid (corresponding to 0.1N) and 10 g of molecular sieve 5A were charged in four flasks, and the pressure was reduced to 60.
Reflux for 8 hours at 0 ° C. After the reaction, the defatted pig skin is filtered,
It was washed several times with 500 ml of acetonitrile. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Example 9 10 g of defatted pork skin, 100 g of glycerin, 1 L of DMF and 2.86 ml of sulfuric acid (corresponding to 0.1N) were placed in an Erlenmeyer flask and shaken at 24 ° C. for 7 days. After shaking, the defatted pig skin was filtered, distilled water was added, neutralized with a 0.1 N sodium hydroxide solution, and washed several times with 500 ml of distilled water. The washed defatted pig skin was treated with an enzyme in a 2% citric acid solution to extract the modified atelocollagen to obtain the desired product. The obtained product was confirmed by the same method as in Example 1. The specific optical rotation of the obtained modified atelocollagen is shown in Table 1.

Comparative Example 1 2.0 g (pure content) of minced and defatted after peeling lime-pickled pig skin was dispersed in 1 L of 0.5 mol acetic acid,
0.05 g of Proctase (manufactured by Meiji Seika Co., Ltd.) was added and the mixture was extracted at 20 ° C. for 72 hours. Extract the solution at 10,000 rpm
Insoluble matter was removed by centrifugation for 30 minutes, 5% sodium chloride was added to the supernatant, and the mixture was allowed to stand for 24 hours to salt out atelocollagen.

Then, the precipitate was recovered by centrifugation at 5000 rpm for 20 minutes and dissolved in 300 ml of 0.5 mol acetic acid.
It was put in a bag of cellulose-based dialysis membrane and dialyzed against 1 L of distilled water. Distilled water was changed every day, and dialyzed for 7 days. After desalting and deoxidizing by dialysis, put in a vacuum container and freeze,
It was dried by a freeze dryer for 7 days to obtain 0.45 g of atelocollagen fiber.

The whole amount was dispersed in 1 L of 99.9% ethanol, 0.01 M of hydrochloric acid was added, and the mixture was stirred at 24 ° C. for 7 days. The modified collagen fiber was filtered off and washed several times with ethanol to obtain 0.4 g of the desired product. In addition, instead of dialysis and freeze-drying, an attempt was made to wash the precipitate of collagen obtained by salting out with ethanol, but it could not be removed because the collagen fiber contained water containing salt. It took 25 days for obtaining ethyl esterified collagen by the above-mentioned process, only from the state of defatted pig skin. Of these, the 14 days required for dialysis and freeze-drying
In the method of the present invention, the period is not necessary but can be shortened.

Comparative Examples 2 to 5 The methyl ester of atelocollagen prepared according to the conventional method of Comparative Example 1 was used as Comparative Example 2, atelocollagen was used as Comparative Example 3, and the one obtained by removing the amide group from collagen was used as Comparative Example 4 ( Table 1 shows the results of measuring the physical properties of the product under the trade name "Desamide Collagen" manufactured by Henkel KK and acid-soluble collagen as Comparative Example 5.

[0045]

[Table 1]

From the results shown in Table 1, the specific optical rotations of the modified collagen of the present invention obtained in Examples 1 to 9 are almost the same as the specific optical rotation of unmodified collagen. It can be seen that the triple helix structure of the modified collagen is maintained.

In order to confirm that the modified collagen of the present invention obtained in Examples 1 to 9 is suitable as a cosmetic base, a solubility test in water and water containing an inorganic salt was conducted.

The solubility test was carried out by adding 1 g of the sample at room temperature to pH.
Water adjusted to 4, 6, 8 or 5% NaCl aqueous solution 99
g and stir to visually check the state of the solution,
The following criteria were evaluated. The results of the dissolution test are shown in Tables 2 and 3 together with the results of unmodified collagen and conventional ester-modified collagen (esterified after extracting atelocollagen from animal tissue).

(Evaluation Criteria for Solubility) ○: Dissolved transparently △: Slightly cloudy ×: Cloudy precipitate (reagent used for pH adjustment) pH 4: citrate buffer (citric acid, sodium citrate) pH 6: phosphate buffer ( NaH ₂ PO ₄ , Na ₂ HP
O ₄ ) pH 8: Glycine buffer (glycine, NaOH)

[0050]

[Table 2]

[0051]

[Table 3]

From the results of Table 2, as compared with the conventional water-soluble collagens of Comparative Examples 1 to 5 which do not dissolve at pH 6 to 8,
The modified collagen of the present invention obtained in Examples 1 to 9 has p
In the range of H4 to H8, it was found that the conventional ester-modified collagen of Comparative Examples 4 and 5 was stably dissolved, and was suitable as a cosmetic base.

From the results shown in Table 3, the modified collagen (ethyl ester or n-dodecyl ester) obtained in Examples 3 and 4 of the present invention was obtained in Comparative Examples 1 and 2 even if an inorganic salt was present. The same degree of solubility as that of conventional ester-modified collagen is exhibited, and further, the modified collagens of Examples 1, 2, 5 to 9 have excellent solubility in a wide pH range of 4 to 8 as compared with conventional modified collagen. Shows. Therefore, it can be seen that the modified collagen of the present invention is suitable as a cosmetic base.

Examples 10 to 12 and Comparative Examples 6 and 7 Lotions having the following formulations were prepared by a usual method, and a sensory evaluation of the feel on the skin was conducted according to the following criteria.
The results are shown in Table 4.

<Formulation of lotion> Ethanol 10.0% by weight Glycerin 3.0 POE (50) oleyl ether 0.5 Methylparaben 0.2 Modified collagen Amount shown in Table 4 Purified water Remaining amount.

<Evaluation of Feeling on Skin> A sensory test was conducted by actually using samples prepared by 20 professional panelists. The evaluation results are shown in the following display.

A: 16 out of 20 responded that they were good. ○: 12 to 15 out of 20 responded that they were good. B: 8 to 11 out of 20 responded that they were good. ×: Less than 8 out of 20 answered that they were good.

[0058]

[Table 4]

[0059]

Industrial Applicability As described in detail above, according to the method for producing esterified collagen of the present invention, it has a wide dissolution range and excellent dissolution stability, and since it can be produced at low cost, its industrial use. The target value is great.

[Brief description of drawings]

FIG. 1 is a photograph showing electrophoresis of unmodified atelocollagen, conventional esterified atelocollagen and modified collagen of the present invention (Example 1).

Front page continuation (72) Inventor Kunio Suzuki 2835 Imafuku, Kawagoe City, Saitama Prefecture Kawaken Fine Chemicals Co., Ltd. Saitama Plant

Claims (5)

[Claims] 1. A method for producing modified collagen, which comprises extracting the modified collagen after modifying the animal tissue containing the collagen. 2. The method for producing modified collagen according to claim 1, wherein the modification is esterification. 3. The alcohol used for esterification is represented by the general formula (1): (In the formula, R ¹ represents a linear or branched alkyl group or alkenyl group having 1 to 20 carbon atoms), a compound represented by the general formula (2): (In the formula, R ² is H, a linear or branched alkyl group having 1 to 20 carbon atoms or an alkenyl group, and n is an integer of 1 to 200); (In the formula, R ³ is H, a linear or branched alkyl group having 1 to 20 carbon atoms or an alkenyl group, and n ′ is an integer of 1 to 200), (In the formula, R ⁴ is H, a hydroxyl group or a linear or branched alkyl group having 1 to 20 carbon atoms, an alkenyl group, an alkoxyl group, Ph is a phenylene group, and n ″ represents an integer of 0 to 200) and glycerin. 3. The method for producing modified collagen according to claim 2, wherein the modified collagen is at least one selected from the group consisting of: 4. The method for producing modified collagen according to claim 1, wherein the animal tissue is at least one selected from cowhide, pig skin and shark skin. 5. A cosmetic base comprising a modified collagen obtained by modifying and then extracting animal tissue containing collagen.