JPS63218607A - Cosmetic compounded with extracted solution of thymus gland - Google Patents

Abstract

PURPOSE:To obtain a cosmetic having strong tyrosinase-suppressing action and high stability, by compounding an extracted solution having a total nitrogen content higher than a specific level and obtained by the hydrolysis of cattle thymus gland. CONSTITUTION:Thymus gland of cattle is used as a starting raw material and is hydrolyzed to obtain an extracted solution. The solution is adjusted to a total nitrogen content of >=0.07w/v% and compounded to a cosmetic. The obtained cosmetic compounded with thymus gland extraction solution has excellent moisture-retaining property and high thermal stability and is effective in preventing the drying of skin and hair to give smooth and soft finished skin and hair and effectively preventing suntan (formation of melanin pigment) by synergistic effect of tyrosinase-suppressing action and ultraviolet absorbing action.

Description

DETAILED DESCRIPTION OF THE INVENTION [A] Object of the Invention The present invention relates to the application of an improved thymus extract solution to cosmetics.

The thymus extract solution used in the present invention is obtained by hydrolyzing bovine thymus gland, and its characteristic is that when quantitatively determined, the solution contains a total nitrogen content of 0.07 w/v% or more. Features.

``Conventional Technology J Extracts obtained from various organs of animals (livestock) such as cows and pigs have been used in pharmaceuticals and cosmetics since ancient times.

Among them, the following method is known as a method for obtaining an animal thymus extract.

(1) There is an antitumor agent (Japanese Patent Publication No. 61-8807) whose active ingredient is an extract mainly consisting of protein obtained by extracting bovine thymus with an aqueous solvent.

In addition, extracts from animal organs used for pharmaceuticals and cosmetics are generally extracted using an extraction solution obtained using an organic solvent, or hydrolyzed using an acid, alkali, or a firefly enzyme. There are extraction solutions obtained by

1. Problems to be Solved by the Invention Recently, obtaining an extract from bovine thymus and applying it to cosmetics has attracted attention in Europe and is well known. However, the extraction method has not been disclosed in detail.

Therefore, the present inventors decided to try applying to cosmetics based on general conventional extraction methods from other organs, and by the method shown in the following table (Table 1), each extraction After obtaining the product, we began our research by confirming the relationship with its action (effect).

As a result, extracts were obtained based on the methods shown in Table 1. As a result, we were able to obtain the results shown in Table 2.

In other words, those obtained using known extraction methods: Sample Arashi 1 (obtained by organic solvent treatment), Sample N112 (obtained by hot water extraction treatment), and Sample No. 1 in Table 1. In order to confirm the effect, three types of extracts (obtained by hydrolysis treatment) of A.
: Respiratory ability (oxygen consumption) on tissue sections; B: Tests were conducted on tyrosinase activity inhibitory effect. The results were different for each.

In other words, it was confirmed that the tyrosinase activity inhibiting effect was strongly exhibited by the extract obtained by hydrolysis, and the effect was superior to that obtained by other extraction methods.

However, when looking at the thermal stability, all of them were unstable.

1 Table 1, Extraction method of thymus extract (Table 2)
Evaluation of the effects of thymus extract'Test method A- in Table 2
Comments on B" [Test method A: Effect of increasing tissue respiration capacity] A test sample was added to a rabbit skin tissue section, and oxygen consumption was determined using a Warburg manometer. Physiological saline was used as a control. In Table 2, all evaluation values for the final method are shown as (+) for those that showed a tendency to increase the # element consumption more than physiological saline.

[Test method B: Tyrosinase activity inhibitory effect and reaction system Derosin solution (L-tyrosine 0.3 / Tsuru) 3 mm, by
77-solution (p with 01M citric acid and 0.2M disodium hydrogen phosphate) (5, g) 3111, and specimen 2.7 m
1 was placed in a constant temperature water bath at 37°C, and left to stand for about 10 minutes. Then, 3 mQ of solution Q, in which tyrosinase was dissolved, was added, and the absorbance of the solution at 475 nm was measured after 15 minutes. Purified water was used as a control. Measured using.

The specimens used in each test method A-BJ were an aqueous solution containing 3% freeze-dried powder of the extract obtained based on the extraction method (known) shown in Table 1. . However, in Test Method A, it was further diluted to prepare an aqueous solution containing 0.02%, and this was used.

Next, based on the results of the effect test shown in Table 2, we consider the following: First, we find that the common effects (effects) of the three types of thymus extracts are similar to those of other organ extracts ( For example, extracts from the liver and placenta).
An effect of increasing tissue respiration capacity was observed. Therefore, extracts that have the effect of increasing respiratory capacity,
As a so-called skin nutrition source (agent), it is useful for preventing aging, gives luster and firmness to the skin, and is considered to be effective as an anti-wrinkle agent. Furthermore, many of the extracts that have tissue respiration ability have the role of alleviating inflammation or acting as a buffer for wounds, etc., and can be used as local tissue regeneration and repair agents.

However, based on the results shown in Table 2, it was found that further improvements were needed in the application of thymus extracts to cosmetics.

That is, in terms of action (effect), it was confirmed that the hydrolyzed extract is superior among the three extraction methods in terms of the inhibitory action on tyrosinase activity. Therefore, using this known extraction method as a base, the present inventors aimed to obtain a thymus gland extract with good stability and even superior action, and to apply this to cosmetics, and conducted research. I was in charge of development.

[Mouth] Structure of the Invention The cosmetic according to the present invention uses bovine thymus as a starting material, and the total nitrogen content of the extracted solution (extract) obtained by hydrolysis is 0.
．． The purpose is to use an extract containing 0.07 w/v% or more.

A means to solve one problem.

When applying thymus extract to cosmetics, there are no other examples of using a solution in which the total nitrogen content of the extracted solution obtained by hydrolysis is 0.07 w/V% or more. I can't find it.

Based on the results shown in Table 2, the present inventors made various improvements in order to obtain a stable and effective thymus extract solution that can be incorporated into cosmetics. However, we conducted research while comparing the effects of the obtained trial extract.

As a result, by using an extraction solution prepared by the method shown in the following example, we succeeded in achieving good stability and a strong tyrosinase activity inhibitory effect.

Example 1: Extraction method The frozen preserved thymus of a fresh cow is crushed with a mince cutter, an organic solvent is added thereto, stirred, dehydrated, defatted, dried, and used as the starting material. Next, about twice the amount of water was added to the starting material and stirred, and then an appropriate acid was added to adjust the pH to 3.0 or less, and the mixture was further heated at 65 to 75°C for 20 minutes.
After heat treatment for ~3 hours, the pH is adjusted to 6-7 with an alkali.

Next, a protease is added to perform hydrolysis (the reaction conditions at this time vary depending on the protease used, but here, protease manufactured by Kaken was used).

After the reaction is completed, the enzyme used is inactivated by heating, and the resulting decomposition liquid is then filtered. Purified water, ethanol, propylene glycol, 1.3
- Add a mixture of one or more of butylene gellicol and chrycerin, and the total nitrogen amount in the solution is 0.07w.
/v% or more, and after adding preservatives,
Again, using a 0.2 μm membrane filter,
Do A and hit the slope! After one treatment, a thymus extract solution is obtained. This is used in cosmetics.

Example 1-2 = Extraction method.

10 kg of cryopreserved thymus of a fresh calf was crushed with a mincing cutter, 10 to 202 n-hexane was added and stirred to transfer the lipids to the n-hexane layer, and the crushed thymus was collected. Dry and use as starting material.

Next, water is added at a ratio of about twice the amount of the starting material,
After stirring, add a small amount of diluted sulfuric acid to adjust the pH to below 3.0, further heat-treat at 65-75°C for 2-3 hours, and then adjust the pH to 6-7 using diurnal sodium hydroxide test solution. Prepare.

Next, a proteolytic enzyme (pronase) is added to perform hydrolysis (reaction conditions include heat treatment at 40 to 60°C for 14 to 8 hours, and after completion of the reaction, the enzyme used is inactivated by heating).

After hydrolysis, filtration is performed, the filtrate is separated, the filtrate obtained here is concentrated under reduced pressure, and lyophilized to form a powder. Purified water alone or purified water is mixed with glycerin or It is dissolved in a mixed solution of propylene glycol and 1,3-butylene glycol, at which time the total amount of nitrogen in the solution is 0.
．． After adding a preservative and filtering again using a 0.2 μm membrane filter and sterilizing, a thymus extract solution is obtained. This is used in cosmetics.

“Example-3 = Extraction method.

After washing fresh calf thymus, mince it with a mince cutter.
After dehydration and defatting, it is dried and used as raw material.

After adding water 201 to raw material 10- and stirring, a small amount of dilute sulfuric acid was added to make the pH 3.0 or less, and the mixture was heated at about 70°C.
After heat treatment for ~3 hours, diurnal cycle - sodium hydroxide test solution,
vI4t to pH 6-7.

Next, a protease (pronase) is added to perform hydrolysis (reaction conditions include heat treatment at 40 to 60°C for 4 to 8 hours, and after the reaction is complete, the enzyme used is inactivated by heating. ).

After hydrolysis, filtration is performed, the filtrate is separated, 2.5 kg of cosmetic primordium/propylene glycol is added, and the mixture is mixed with stirring. At that time, the total amount of nitrogen in the solution is 0.08~
Prepared to have a concentration of 0.25 w/v%, then 11
7JjIl! As an agent, makeup primordium-potassium sorbate 0
．． 1%, cosmetic primordium/methyl paraoxybenzoate 0.17%
, cosmetic primordium/propyl paraoxybenzoate 0.015%,
After adding and dissolving 0.015% of butyl paraoxybenzoate, the mixture is refiltered using a 0.2 μm membrane filter and sterilized to obtain a thymus extract solution.

This is used in cosmetics.

The combination of preservatives specified in "Example-3" does not cause irritation and improves the stability of the solution.
It also has a positive effect on the stability of cosmetics using it.

In addition, water-soluble low-molecular-weight peptides have migrated into the extraction solution obtained in the example, and RNA +
DNA, ATP, etc. have also migrated considerably. Moreover, when looking at the molecular weight distribution state, although it differs depending on the temperature and time during hydrolysis, for example, according to Example 3, there is a peak around the molecular weight of 100 to 300, a peak centered around around 500, and A product with a peak around 1,000 to 5,000 is obtained.

1. Confirmation of the effect (effect! Ta.

The results are shown in Table 3, approximately 0.07w
It was found that the effect was reliably observed at a mixing ratio of /v% or higher (i11 degrees).

From the results of the evaluation test (Table 3) regarding the effect of thymus extract solution, the extract solution-containing aqueous solution obtained in the example shows a good tyrosinase activity inhibiting effect in a small amount. .

That is, the optimal effect (
Regarding the effect), from the main quantitative values of the contained component composition,
When the total nitrogen amount was determined, it was found to be 0.07 w/v% or more, indicating that both respiratory ability and tyrosinase activity inhibitory effect were good, and thermal stability (color change) was also reduced. In other words, it was found that the solution was excellent for use in cosmetic formulations.

Furthermore, when the ultraviolet absorption spectrum of the extracted solution obtained in the example was determined, it was as shown in FIG. 1, and it was confirmed that the ultraviolet absorption spectrum had a peak around about 240 to 28 Onm.

On the other hand, from the components of the extraction solutions obtained in the examples, as shown in Table 4, by specifying them based on the total nitrogen content, it is possible to create an extraction solution that is superior to conventional animal-derived extraction solutions. It is possible.

Next, looking at the moisturizing effect of the extract solution obtained in the example, it is as shown in FIG.

In addition, the test method was to determine the moisture loss rate when the samples were allowed to stand for 16 hours at relative humidity of 20%, 40%, 60%, and 80% at a temperature of 30°C using a constant temperature and humidity machine (manufactured by Takasugi Seisakusho). It is something.

Based on the extraction method described in the example, the final yield obtained from 1 kg of the starting material thymus (unprocessed, before drying) is the minimum yield when converted to the freeze-dried powder of the extract. Approximately 60g was obtained, with a maximum of 180g.

On the other hand, Table 4 below shows the physical properties of the extraction solutions obtained in the examples. 'Improved by adding tylene glycol. Particularly, when the extraction solution contains 0.04% or more as a total nitrogen content, transparent liquid properties are maintained by adding these polyol solvents. Therefore, in lotions and the like, it is desirable to use these polyol solvents together.

"Table 4. Standards or physical properties of thymus extract solution Next, regarding the stability of each extraction solution, we look at changes over time at low temperatures to 40°C for one month, as shown in Table 5. It is.

That is, the extraction solution according to the present invention has excellent thermal stability. The reason for this is thought to be that the treatment during the extraction process is important.In the final method, as a pretreatment to obtain the extraction solution, drying is performed along with defatting and blood removal, and then the enzyme nib-a-nase is used. During hydrolysis, heat treatment using dilute sulfuric acid is performed, and this operation greatly contributes to thermal stability.

1 Table 5 Evaluation of thermal stability 1 Formulation example-1: Lotion Extract solutions obtained in Examples 1 to 3 (however, the extract solution was prepared to contain 0.5 w/v% as a total nitrogen amount) solution)··
...10 to 30% Nuconic liquid AB (solution containing pigment extracted from Niconia) ...0.3 to 0
, 5% 1.3-budelene glycol 2-6%
Fragrances and preservatives... Appropriate Make up to 100 with water at the end.

1 Prescription example-2: Emulsion.

Extraction solutions obtained in Examples 1 to 3 (however, the total nitrogen content was adjusted to 0.5 w/v%)...
...t...3-20% stearic acid...
... 2.5% cetyl alfur... --■ ---1
, 5% white petrolatum... 5.0%
Liquid paraffin 10.0% Twin 80 1 2.0% polyethylene glycol (1500) 3.0% triethanolamine ... 1.0% fragrance and helmet 1...
・・・・・・ Take the water at the end and make it ioo.

'Formulation Example-3: Cream' Extract solutions obtained in Examples 1 to 3 (solutions prepared to contain Q, 5 w/v% as total nitrogen content)...
...3-6% polyoxyethylene oleyl ether, 1.6% lanolin alcohol...T1--
1.8% lanolin 5.4
%Setanol・・・・・・・・・ 2.0% Bioderma SX-19 (Navatar)・9.0% Liquid paraffin・・・・・・・・・ 6.0% Monostearic acid Glycerin...-2.0% Oryza oil 5-1 (rice germ oil)...4.0% Isopropyl palmitate...
・・10.0% Ceresin ψ・・・KomaΦ・φ・Excellent・
2.0% borax... Conflict...◆... 1.
0% fragrance and preservatives... Suitable Add water to the end and make it into a zoo.

[C] Effects of the Invention The present invention provides an extraction solution according to the present invention characterized in that the total nitrogen content of the obtained extraction solution by hydrolyzing the thymus gland is 0.07 w/v% or more. The extract according to the present invention contains a large amount of water-soluble low-molecular-weight peptides and amino acids, and also contains RNA and DNA% ATP. When combined with this product, it prevents the skin and hair from drying out, leaving them moist, smooth, and flexible.

Furthermore, as shown in Table 3, it has a tyrosinase activity inhibitory effect and is expected to suppress the production of colored melanin in the skin.

That is, the characteristics of the extract according to the present invention are that the optimum concentration of the extract is determined with a focus on the activity inhibition effect on delobnase, and a solution containing 0.07 w/v % or more as a total nitrogen content in the extraction solution is used. There is a particular thing. Therefore, the concentration is the same as the amount (concentration) of cosmetics, and the purpose is to suppress the production of colored pigments on the skin. As a guideline for the amount of nitrogen added, when quantifying the total amount of nitrogen derived from the extract, it is desirable that the content be 0.07 w/v% or more.

Traditionally, many organ-derived extracts have been used in cosmetics, but
Until now, there have been few cases in which the relationship between the optimal amount has been determined based on the amount added and the ability to suppress tyrosinase activity, and the present inventors have discovered this relationship for the first time, especially for thymus-derived extracts.

In addition, as shown in 1st rg: J, it has ultraviolet absorption ability,
Therefore, it can be said that it helps protect the skin from direct sunlight and has great cosmetic effects.

In other words, if the extract of the present invention is used, it will absorb ultraviolet rays and produce tanning (melanin pigment production), while simultaneously directly suppressing tyrosinase activity (synergistic effect). It is something that we have.

On the other hand, when used in cosmetics, extracts derived from organs generally have the disadvantage of poor thermal stability, and along with this, the optimum concentration (0.07 w/v) as mentioned above is required. %
However, as shown in Table 5, stability is desirable from the formulation standpoint, and it should be noted that heat treatment is not required for creams and milk products. There are many, but in 2 to 3 hours,
There were no problems with stability, and even when the heat-treated extract was tested for its tyrosinase activity inhibitory effect, the effect was maintained and no decrease was observed.

This is a great advantage when applied to cosmetics, and can be used in a wide range of applications, from water-soluble cosmetics to emulsion and dispersion cosmetics, as well as powder and solid cosmetics. It means that it is possible. However, when compounding into cosmetics, preservatives,
The selection of the anti-inflammatory agent is important, and for example, the combination of each agent shown in Example 3 is good.

[Brief explanation of the drawing]

FIG. 1 is an ultraviolet absorption spectrum of an aqueous solution containing 5% thymus extract obtained according to the present invention. FIG. 2 is a graph showing the moisturizing effect of the thymus extract solution (containing 0.5% total nitrogen content) obtained according to the present invention.

Claims (1)

[Claims] (1) A cosmetic comprising a solution obtained by hydrolyzing bovine thymus as a starting material and containing an extracted solution containing a total nitrogen content of 0.07 w/v% or more.