JPH1179934A - Tyrosinase activity inhibiting composition, tyrosinase activity inhibitor, and bleaching solution and bath preparation using the same composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a tyrosinase activity inhibiting composition, which prevents sunburn-caused pigmentation or blotches/freckles, and also to obtain the bleaching solution or bath preparation using the same composition. SOLUTION: This composition contains, as the active ingredients, a low- molecular-weight thiol compound, e.g. thiodiglycolic acid or thioctic acid, and a low-molecular-weight thiol compound having a molecular weight of 500 or less, found in sake lees. The skin-whitening solution or bath preparation uses the same composition.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel composition which inhibits the activity of tyrosinase which prevents pigmentation of the skin due to sunburn and the occurrence of spots and freckles, and a whitening serum or whitening bath using the same. Concerning the realization of

[0002]

2. Description of the Related Art Tyrosinase is one of metal enzymes widely existing in microorganisms, plants, animals, etc., and plays an important role in catalyzing the oxidation of tyrosine and synthesizing melanin. I have. In humans, melanin synthesis is carried out by melanocytes in the basal layer of the epidermis by the action of tyrosinase induced by ultraviolet rays (G. Prota, J. et al.
Invest, Delmatol. 75, 122-127, 1980).
By inhibiting ze activity, pigmentation of the skin due to sunburn and occurrence of spots and freckles can be prevented. Therefore, various substances having an effect of inhibiting tyrosinase activity have been studied in various fields. At present, Tyrosina
Quasi-drugs such as kojic acid, arbutin, and ascorbic acid derivatives have been used as zein activity inhibitors. However, no tyrosinase activity inhibitor is satisfactory because of lack of stability, difficulty in obtaining raw materials, or being very expensive. Therefore, when we searched for cheaper raw materials that were easily available, we found a very strong inhibitory effect on tyrosinase activity in the organic solvent solubles of the rice fermented product. And found that a tyrosinase activity inhibitor was present in the sake lees (Japanese Patent Application Laid-Open No. 6-199679).

[0003]

However, since it was unknown at this time what tyrosinase activity inhibitor was present in sake lees, liquid-liquid partitioning, various types of chromatography, etc. were performed to identify the substance. Was repeated, and the basic properties of the tyrosinase activity inhibitor were examined. As a result, they were found to be low-molecular-weight thiol compounds. Then, further research was conducted, and it was investigated and confirmed whether or not a readily available specific low molecular weight thiol compound has the property of inhibiting tyrosinase activity. As a result, it was found that not only low-molecular-weight thiol compounds such as thiodiglycolic acid and thioctic acid, but also low-molecular-weight thiol compounds derived from sake lees, in particular, have the property of strongly inhibiting tyrosinase activity. Was. Therefore, based on this new finding, the present inventors have embodied a composition that inhibits the activity of tyrosinase with easily available and inexpensive raw materials, and a skin whitening serum or whitening bath agent using the same. Things.

[0004]

The first invention to be patented is a composition for inhibiting tyrosinase activity, which comprises a low molecular weight thiol compound as an active ingredient.

[0005] The second invention to be patented is characterized in that the low molecular weight thiol compound as an active ingredient is any of thiodiglycolic acid and thioctic acid.
A tyrosinase activity inhibiting composition according to the invention.

A third invention which is to be patented is characterized in that it comprises a low molecular weight thiol compound having a molecular weight of 500 or less contained in sake lees as an active ingredient.
It is a zease activity inhibiting composition.

[0007] The fourth invention to be patented includes thiodiglycolic acid, thioctic acid and low molecular weight thio-acid derived from sake lees.
A tyrosinase activity-inhibiting composition comprising at least one active compound as an active ingredient.

The fifth invention for which a patent is sought includes thiodiglycolic acid, thioctic acid and low molecular weight thio-acid derived from sake lees.
A tyrosinase activity inhibitor comprising at least one compound as an active ingredient.

The sixth invention for which a patent is sought includes thiodiglycolic acid, thioctic acid, and low molecular weight thio-
A whitening essence characterized by comprising at least one active ingredient among the compounds.

[0010] The seventh invention for which a patent is sought includes thiodiglycolic acid, thioctic acid, and low molecular weight thio-acid derived from sake lees.
A whitening bath agent comprising at least one active compound as an active ingredient.

[0011]

Next, the present invention will be described in detail with reference to examples. First, an experiment was conducted to examine the tyrosinase activity inhibiting effect of the tyrosinase activity inhibiting composition derived from sake lees.

1. Preparation of composition for inhibiting tyrosinase activity derived from sake lees In order to use sake lees having a stronger tyrosinase inhibitory activity as a starting material, the tyrosinase inhibitory activities of several types of obtained sake lees were measured. An 80% ethanol solution of sake lees was thoroughly stirred with a homomixer, allowed to stand, and the ethanol layer was concentrated to obtain an ethanol-soluble fraction, which was used for activity measurement. Tyrosinase is derived from mashroom (manufactured by Sigma,
3.000 units / mg solid) in 0.1 M phosphate buffer (p
H 6.8), adjust to a concentration of 800 units / ml, and
-Tyrosine (special grade, manufactured by Wako Pure Chemical Industries) is 1.66μ in the same buffer
It dissolved so that it might become mol / ml. Macilva
ine) 1 ml buffer (pH 6.8) and 0.1 ml sample, tyrosinase 0.
1 ml of the mixture was mixed and pre-incubated at 37 ° C. for 10 minutes. Then, 1 ml of the substrate solution was added and reacted at 37 ° C. for 90 minutes. The inhibitory activity was determined by measuring the absorbance at 490 nm using a spectrophotometer.

[0013] As a result, purification was proceeded using the most active sake lees as a starting material. Dichloromethane was added to the ethanol-soluble fraction of the sample, the dichloromethane layer was separated, and extracted with water. The obtained aqueous layer was treated with activated carbon, and the fraction not adsorbed with activated carbon was concentrated and freeze-dried. Next, Pharmacia's Superdex Peptide column (Superdex Peptide co
The purified product was subjected to gel filtration using (lumn) (1.0 × 30 cm) (FIG. 1). The solvent is distilled water and the flow rate is 1.0 ml / min,
The absorption of the sample at 260 nm was monitored. Since a tyrosinase inhibitory activity was observed only in the first peak at a retention time of about 17 minutes, the peaks were collected, rechromatographed on the same column (FIG. 1), and a tyrosinase activity inhibiting composition derived from sake lees. I got

2. Properties of the tyrosinase activity inhibitory composition derived from sake lees When the mass spectrum of the tyrosinase activity inhibitory composition derived from the sake lees obtained by the above preparation was directly measured by the FAB method, several signals were obtained at a molecular weight of 500 or less. The tyrosinase activity inhibiting composition has a molecular weight of 500
The following low molecular weights were suggested. In addition, the results of IR showed the presence of a thiol group (FIG. 2).

Further, tyrosinase 8 derived from mashroom
For 00 units, tyrosinase derived from sake lees - IC ₅₀ of peptidase activity inhibiting composition (ug / ml) in 2.5, low molecular thio were compared - stronger than Le compounds tyrosinase - peptidase inhibitory activity was observed (Table 1). In addition, Lineweaver Burke
aver-Burk) Tyrosine derived from sake lees by plot method
Examination of the type of inhibition of the zein activity inhibitory composition revealed a non-antagonistic type (FIG. 3).

3. Tyrosine, a low molecular thiol compound
Inhibitory activity on ze. Since the composition for inhibiting tyrosinase activity derived from sake lees had a thiol group (FIG. 2), the inhibitory activity on tyrosinase of a readily available low-molecular-weight thiol compound was confirmed. The inhibitory activity of thiodiglycolic acid (manufactured by Wako Pure Chemical Industries, Ltd.) and thioctic acid (manufactured by Sigma), which is a kind of vitamin B group, as low molecular thiol compounds on tyrosinase was examined. Tyrosinase is mash-
Originated from Sigma (3,000 units / mg.s
olid) in a 0.1 M phosphate buffer at 800 units / ml
What was adjusted so that it might become. The IC ₅₀ (ug / ug) of the low molecular weight thiol compound with respect to 800 units of the tyrosinase is used.
ml), thiodiglycolic acid was 9.8, and thioctic acid was 14.7, all of which exhibited inhibitory activity against tyrosinase (Table 1). In addition, the inhibition forms of thiodiglycolic acid and thioctic acid were all non-antagonistic. As a result, it was found that both can be used as a tyrosinase activity inhibiting composition.

[0017]

[Table 1]

4. Next, the present inventors tried to produce a serum having a high whitening effect by suppressing the synthesis of melanin by the action of tyrosinase. A serum containing 1% (W / V) of the composition for inhibiting tyrosinase activity derived from sake lees described in the above 1 and having the following components was prepared. Ten adult women continued to use their face and hands twice daily in the morning and evening during the three months in the summer to examine their whitening effect. As a result, it was confirmed that even after the elapse of the period, almost no or little pigmentation of the skin due to sunburn was observed for all the subjects. In addition, five adult women who were concerned about spots and freckles on the face were used twice daily in the morning and evening for 3 months, using them on their faces and hands, observing changes in the condition of the freckles and whitening. The effect was investigated.
As a result, it was confirmed that the color of the spots and freckles became light.

<Blending of whitening beauty liquid> (%) Tyrosinase activity inhibitor derived from sake lees 1.00 Wetting agent 11.00 Solubilizing agent 0.45 Stabilizing agent 0.04 Moisturizing agent 0.09 Solvent (ethanol) 4.96 Purified water 82.46

[5] Preparation of Whitening Bath Agent and Whitening Effect Next, the whitening bath agent was used for the purpose of whitening the whole body.
A serum containing 10% (W / V) of the tyrosinase activity-inhibiting composition derived from sake lees described in the above 1 and containing the following components was prepared. 1 daily bath for 10 adult women
We had you use for months and looked at whitening effect. As a result, it was confirmed that pigmentation of the skin due to sunburn was prevented, and the spot color of the body was lightened.

<Blending of whitening bath agent> (%) Tyrosinase activity inhibitor derived from sake lees 10.0 Solvent 20.0 Solubilizer 5.0 Wetting agent (whey mineral powder) 0.5 Flavoring agent 0. 9 Purified water 63.6

[0022]

As described above, a tyrosinase activity-inhibiting composition containing a low-molecular-weight thiol compound as an active ingredient, such as thiodiglycolic acid and thioctic acid, has a tyrosinase activity. Has the effect of inhibiting.

In addition, the composition for inhibiting tyrosinase activity, which contains a low molecular weight thiol compound having a molecular weight of 500 or less contained in sake lees as an active ingredient, also inhibits a very strong tyrosinase activity. Has the effect of doing

Therefore, using a low molecular weight thiol compound, thiodiglycolic acid, thioctic acid, or a low molecular weight thiol compound derived from sake lees, a tyrosinase activity inhibitor containing the same as an active ingredient was prepared. However, they have the effect of inhibiting potent tyrosinase activity.

Further, low molecular weight thiol compounds such as thiodiglycolic acid, thioctic acid, and low molecular weight thiol
Whitening bath preparations and whitening essences containing a compound such as a compound as an active ingredient have been produced, which have a whitening effect.

[Brief description of the drawings]

BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a graph showing a gel filtration pattern of a refined sake lees by a column and a pattern obtained by rechromatizing a main part thereof by the column.

FIG. 2 is a measurement graph of an IR spectrum of a composition for inhibiting tyrosinase activity derived from sake lees.

FIG. 3 is a graph showing an inhibition state of a mash-derived tyrosinase activity-inhibiting composition with respect to mashroom-derived tyrosinase activity.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 7/50 A61K 7/50 31/095 31/095 31/19 ADA 31/19 ADA 31/385 AED 31/385 AED 35 / 78 35/78 U C12N 9/99 C12N 9/99 (72) Inventor Hisanori Tani 6-14-7 Akikawa, Akiruno-shi, Tokyo (72) Inventor Ichizo Oishi 3-15-24 Nishisunacho, Tachikawa-shi, Tokyo

Claims (7)

[Claims] 1. A composition for inhibiting tyrosinase activity, comprising a low molecular weight thiol compound as an active ingredient. 2. The tyrosinase activity-inhibiting composition according to claim 1, wherein the low molecular weight thiol compound as an active ingredient is any of thiodiglycolic acid and thioctic acid. 3. A composition for inhibiting tyrosinase activity, comprising a low molecular weight thiol compound having a molecular weight of 500 or less contained in sake lees as an active ingredient. 4. A tyrosine containing at least one of thiodiglycolic acid, thioctic acid and low molecular weight thiol compounds derived from sake lees as an active ingredient.
Zease activity inhibiting composition. 5. A tyrosine comprising at least one of thiodiglycolic acid, thioctic acid, and low-molecular-weight thiol compounds derived from sake lees as an active ingredient.
Activity inhibitor. 6. A whitening essence comprising at least one of thiodiglycolic acid, thioctic acid, and low molecular weight thiol compounds derived from sake lees as an active ingredient. 7. A whitening bath agent comprising as an active ingredient at least one of thiodiglycolic acid, thioctic acid, and low molecular weight thiol compounds derived from sake lees.