JPH0867617A - Cosmetic - Google Patents

Abstract

PURPOSE: To obtain a cosmetic high in whitening activity, capable of inhibiting the activity of hyaluronidase, high in collagenase activity-inhibitory effect, and effective for preventing skin chapping, containing, as active ingredient, a solvent extract from Terminalia horrida used as a crude drug in Egypt. CONSTITUTION: This cosmetic contains, as active ingredient, an extract derived from Terminalia horrida used as a crude drug in Egypt by subjecting the plant to extraction with water or a hydrophilic organic solvent such as ethanol or a mixed solvent containing ethanol. This extract presents skin whitening activity, hyaluronidase activity-inhibitory effect, active oxygen-inhibitory activity, antioxidative effect, antiplasminic activity, and collagenase activity-inhibitory effect. This extract can also be used as powder after lyophilized. This cosmetic can be formulated to various kinds of preparations such as lotion, cream, milky lotion or pack.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cosmetic composition which has a high whitening effect, inhibits the activity of hyaluronidase, has a strong inhibitory effect on collagenase activity, and is effective for rough skin.

[0002]

[Prior Art] Terminaria Horida (Scientific Name: Termin
alia horrida) is a plant of the genus Momotamana,
Used as a crude drug in Egypt.

On the other hand, collagen is also called collagen and is a protein that constitutes connective tissue of animal cells and is widely distributed in the living body. It is considered to exist in multicellular animals, and at least 11 kinds of molecules have been discovered to date, and these genes have different genes from each other.

Collagen, which is the main component of the skeletal structure, has a platelet aggregation action and is also involved in thrombus formation. It has also been suggested that collagen is involved in liver cirrhosis and arteriosclerosis.

Collagenase refers to an enzyme produced by animal tissue cells, inflammatory cells, tumor cells and the like, which decomposes type I, type II and type III collagen. Elucidation of proteins that inhibit collagen degradation
h) is a metalloprotease inhibitor protein (TIMP)
The gene was isolated from human tissue and successfully cloned. Metalloprotease is an enzyme that decomposes collagen (a kind of collagenase), and TIMP inhibits the activity of this enzyme. This TIMP is expected to be an effective therapeutic drug for arthritis, cancer, and skin lesions.

The collagenase inhibitor means a substance which inhibits the activity of collagenase as described above. In the process of tissue destruction and repair during pathology, abnormalities in collagenase activity are observed. These conditions include rheumatoid arthritis,
There are periodontitis, corneal ulcer, congenital epidermolysis bullosa, etc. Therefore, it is considered that the collagenase inhibitor prevents or cures the deterioration of these pathological conditions by inhibiting the collagenase activity.

As a prior art relating to a collagenase inhibitor, JP-A-1-222782 discloses a collagenase inhibitor characterized in that a solution of a collagenase inhibitor is brought into contact with a red dye-binding carrier or a blue dye-binding carrier to elute the collagenase inhibitor solution. A law has been proposed. This is just a method for purifying collagenase inhibitors.

Further, Japanese Patent Laid-Open No. 4-44331 discloses that
A collagenase inhibitor comprising an extract extracted from cacao husk, which is a soybean hull of cocoa beans, is disclosed. In terms of a collagenase inhibitor consisting of an extract extracted from a natural product, there is a part common to the present invention,
Clearly, the natural product itself is quite different.

Further, Japanese Patent Publication No. 2-502537 discloses a synthetic inhibitor of mammalian collagenase. This is to prevent the synthesis of collagenase in the mammalian body and is a thiol-containing peptide, which is different from the present invention in that it is a synthetic product.

On the other hand, various substances are known as substances having a whitening effect which can be used as a raw material for cosmetics, but synthetic products have no guarantee of safety when applied to human skin for a long period of time. , Its use is being limited. In addition, many natural products have a weak whitening effect. However, from the viewpoint of safety for human skin, it is a natural product, and people eat it for many years,
Guaranteed in terms of safety and strong whitening effect,
Furthermore, a substance that also has other effects on the skin has been desired.

[0011]

DISCLOSURE OF THE INVENTION The object of the present invention is that it is safe to be applied to the skin, has a large whitening effect and inhibits the activity of hyaluronidase, has a strong inhibitory effect on collagenase activity, and is effective for rough skin and the like. To provide cosmetics containing ingredients.

[0012]

[Means for Solving the Problems] In order to solve the above-mentioned problems, the present inventors have screened and investigated plants that have been used for food for many years and have been confirmed to be safe for the human body. I examined what is useful as. As a result, Terminaria Horida (scientific name:
The present invention was completed by discovering that Terminalia horrida) is effective as a raw material for cosmetics or as a quasi drug. That is, the cosmetics of the present invention are Terminalia horri (scientific name: Terminalia horri
da) containing the solvent extract.

[0013]

[Action] The confirmed action of the solvent extract of Terminalia horrida (scientific name: Terminaliahorrida) used as the cosmetics of the present invention is to whiten the skin, inhibit hyaluronidase activity, inhibit active oxygen, antioxidative action, It has antiplasmin action and collagenase activity inhibitory action. The activity suppressing effect of the hyaluronidase will be described in more detail. Hyaluronidase is an enzyme that is widely distributed in the living body and is also present in the skin. As its name implies, it decomposes hyaluronic acid. Hyaluronic acid is a linear polymer polysaccharide in which β-D-N-acetylglucosamine and β-D-glucuronic acid are alternately bound, and is a glycosaminoglucan that widely exists in connective tissues of mammals along with chondroitin sulfate and the like. Is a kind of. The action of hyaluronic acid in connective tissue is to retain water in the intercellular spaces, form a jelly-like matrix in the tissue to retain cells, maintain the lubricity and flexibility of the skin, and apply external force ( Mechanical damage) and bacterial infections. In addition, it is said that hyaluronic acid in the skin decreases with age, resulting in aging such as wrinkles and bulkiness.
Therefore, suppressing the activity of hyaluronidase that decomposes this hyaluronic acid, the stability of hyaluronic acid used in the formulation, the hyaluronic acid of the formulation after application to the skin and the hyaluronic acid present in the skin It is thought to contribute to stability.

The effect of suppressing the active oxygen will be described in more detail. Generally, no organisms (except anaerobic ones) can exist without oxygen in the air. However, oxygen becomes active oxygen under the influence of ultraviolet rays and enzymes. This active oxygen oxidizes a fatty acid and produces a peroxide. It also oxidizes and damages the phospholipids of biological membranes in the body. Furthermore, it is said that the generated peroxide and active oxygen damage DNA and accelerate aging. This active oxygen is
It also affects the mechanism of melanin production from tyrosine and is involved in skin darkening. Suppressing this active oxygen is an important factor for the skin, in other words, an important factor required for cosmetics.

Further, the above antiplasmin action will be described in more detail. Normally, erythema and edema are formed by irradiating the skin with ultraviolet rays (UVB) in an amount twice the minimum amount of erythema.
It has been reported that histamine, serotonin, prostaglandin E ₂ , F _2α quinine and the like are involved as chemical mediators in this UV inflammatory reaction. In addition, after UV irradiation, the plasmin activity in rabbit skin tissues was enhanced, the plasmin activity varied well in parallel with the degree of swelling of the skin, and the administration of the antiplasmin agent tranexamic acid caused swelling of the skin. Is known to be severely suppressed. Therefore, the antiplasmin agent is also one of the important chemical mediators in the UV inflammatory reaction, and suppressing this plasmin is an important factor for the skin, in other words, an important factor required for cosmetics.

Furthermore, the collagenase activity inhibitory action will be described. As described above, tissue destruction during pathological conditions,
Since abnormalities in collagenase activity are observed in the repair process, inhibition of this collagenase activity is important for the skin, in other words, an important factor required for cosmetics.

The details of the present invention will be described below.
In the present invention, Terminaria Horida (scientific name: Term
Inalia horrida) is extracted with water or a hydrophilic organic solvent such as ethanol, methanol, acetone or the like. However, it is natural that extraction with water, ethanol, or a mixed solvent thereof is preferable because it is extraction of cosmetic raw materials. In some cases, a polyhydric alcohol such as glycerin, 1,3-butylene glycol, propylene glycol or a mixed liquid of polyhydric alcohol and water can be used for extraction. Furthermore, freeze-drying and using it as a powder is also effective depending on the method of use.

In the cosmetic of the present invention, this solvent extract is used as another cosmetic raw material, for example, liquid oil such as squalane and jojoba oil, solid oil such as beeswax and cetyl alcohol, various active agents, glycerin, 1,3. -Cosmetics of various dosage forms can be prepared by adding moisturizing agents such as butylene glycol and various agents. For example, a lotion, a cream, a milky lotion, a pack or the like can be prepared into cosmetics of various usage forms according to the purpose.

[0019]

[Examples] Hereinafter, production examples of the extract of Terminalia horrida (scientific name: Terminalia horrida) used in the present invention and examples of actual utilization will be described. The present invention, however, will be described. It is not limited in any way by example.

[Production Example 1] 300 g of ethanol was added to 10 g of the fruit (dry product) of Terminaria horrida and left for 5 days with occasional stirring. This was filtered, evaporated and freeze-dried.

[Production Example 2] 300 g of 50% ethanol was added to 10 g of the fruit (dry product) of Terminaria horrida, and the mixture was left for 5 days with occasional stirring. This was filtered, evaporated and freeze-dried.

[Production Example 3] 300 g of purified water was added to 10 g of the fruit (dry product) of Terminaria horrida and heated for 3 hours. This was allowed to cool, then filtered and freeze-dried.

[Example 1 (Preparation of lotion)] The following components were mixed to prepare a lotion by a conventional method. (Wt%) Olive oil 0.5 Ethanol extract of Terminaria horrida of Production Example 1 0.5 Polyoxyethylene (20 E.O.) sorbitan monostearate 2.0 Polyoxyethylene (60 E.O.) hydrogenated castor oil 2.0 Ethanol 10.0 1.0% sodium hyaluronate aqueous solution 5.0 Purified water 80.0

[Example 2 (Preparation of cream)] A and B each consisting of the following ingredients were heated to 70 ° C., and then A was slowly added to B with stirring and then slowly stirred. While cooling to 30 ° C, a cream was prepared. (Wt%) A squalane 20.0 Olive oil 2.0 Mink oil 1.0 Jojoba oil 5.0 Beeswax 5.0 Cetostearyl alcohol 2.0 Glycerin monostearate 1.0 Sorbitan monostearate 2.0 Production example 2 50% ethanol extract of Terminaria horrida of 1.0 B Purified water 47.9 Polyoxyethylene (20 E.O.) sorbitan monostearate 2.0 Polyoxyethylene (60 E.O.) hydrogenated castor oil 1.0 Glycerin 5.0 1.0% sodium hyaluronate aqueous solution 5.0 Methyl paraoxybenzoate 0.1

[Example 3 (Preparation of lotion)] The extract of Production Example 1 in Example 1 was replaced with the extract of Production Example 3 to prepare.

[Inhibition of tyrosinase activity] (Test method) McClevaln buffer solution 0.9 m
l, 1.66 mM Tyrosine solution 1.0 ml, 0.1 wt / v% aqueous solution of each of the above production examples (freeze-dried product) (if it is difficult to dissolve, add ethanol to dissolve it, and then add purified water to evaporate After removing the ethanol,
1.0 ml (prepared to be 0.1 wt / v%) was placed in a screw vial and heated in a 37 ° C. constant temperature water bath for 5 minutes or more. 0.1 ml of a tyrosinase solution (Sigma, mushroom-derived, 914 units / ml) was added, and the mixture was kept warm in a constant temperature water bath at 37 ° C., and after 10 minutes, the absorbance was measured at 475 nm.
As a control, pure water was added instead of the sample solution, and the measurement was performed in the same manner. In this test, the final concentration of the sample is 0.033%. (Calculation formula) Tyrosinase activity inhibition rate (%) = {B- (AP)} /
B × 100 However, A: Absorbance of sample specimen B: Absorbance of control P: Absorbance due to coloring of sample specimen (3-fold dilution)

[0027]

[Table 1]

[Hyaluronidase activity inhibition test] (Test method) 0.4% sodium hyaluronate 0.1M
(PH 6.0) 6 g of phosphate buffer solution was weighed and allowed to stand in a constant temperature water bath at 37 ° C. for 5 minutes, and then 0.1 wt / v% aqueous solution of each of the above production examples (freeze-dried product) Was added and dissolved, purified water was added to the residue to evaporate it, and ethanol was removed. Then, the solution was adjusted to 0.1 wt / v%), and 1.0 ml was added and stirred to prepare 0.01% hyaluronidase (Sigma). Beef testicles, type I-S)
1 ml of 0.1 M (pH 6.0) phosphate buffer was added and immediately stirred, and 6 ml was placed in an Ostwald viscometer placed in a 37 ° C. constant temperature water bath. The viscosity was measured after 5 minutes, 10 minutes, 20 minutes, and 40 minutes. Further, as a control, pure water was added instead of the sample solution, and the same measurement was performed. In this test, the final concentration of each sample is 0.0125% of the concentration of the analyte. The viscosity after 1 minute is 100,
The respective results are shown in Tables 2 and 3 below as indexes.

[0029]

[Table 2]

[0030]

[Table 3]

[Active Oxygen Suppression Test] Suppress active oxygen
There are various methods to measure the effect, but this time the following method is used.
I used it. pH 7.8 50 mM potassium phosphate buffer (containing 1.3 mM DETAPAC) 133 ml 40 unit / ml Catalase above potassium phosphate buffer 5 ml 2 mM nitroblue tetrazolium above potassium phosphate buffer 5 ml1.8 mM xanthine potassium phosphate buffer above 17 ml 160 ml

2.4 ml of the mixture of the above reagents and 0.3 ml of the sample
In addition, a solution of xanthine oxynase (prepared with water as a sample in advance and adjusted with the above-mentioned potassium phosphate buffer solution so that the absorbance increases by about 0.02 per minute) is used.
Add 1 ml and measure the absorbance (560 nm) immediately (measurement is 2
Quantile and confirm linearity). (Calculation formula) Inhibition rate = {(A−B) / A} × 100 where A: change in absorbance per minute when the sample is water B: change in absorbance per minute of sample Several steps were carried out to find the 50% active oxygen production inhibitory concentration. The preparation method of the sample is the above manufacturing example (freeze-dried product)
An aqueous solution having an appropriate concentration was prepared (if it is difficult to dissolve, ethanol was added to dissolve it, and purified water was added to the solution to evaporate it to remove ethanol, and then an appropriate concentration was prepared). The results are shown in Table 4 below.

[0033]

[Table 4]

[Antioxidant test] 50 with the following screw cap
ml test tubes were made. Specimen 5 mg 2% linoleic acid ethanol solution 10 ml 0.1 M, pH 7.0 phosphate buffer 10 ml Purified water 5 ml This 50 ml test tube with a screw cap was left in a thermostat bath at 50 ° C in the dark. The following measurements were performed before putting this in a constant temperature bath and at intervals of several days. After adding 0.125 ml of the test solution, 12.125 ml of 75% ethanol, and 0.125 ml of 30% ammonium thiocyanate, the mixture was stirred and allowed to stand for 3 minutes, and then added to 0.
02N ferrous chloride 3.5% HCL aqueous solution 0.125 ml
Was added, and the mixture was stirred and left standing for 3 minutes, and then the absorbance was measured at a wavelength of 500 nm. The cell length was 10 mm, and the control cell had the test solution replaced with water. The results are shown in Tables 5 and 6 below.

[0035]

[Table 5]

[0036]

[Table 6]

As is clear from the results of Tables 5 and 6, it was found that the antioxidant activity is higher than that of vitamin E currently used as an antioxidant.

[Antiplasmin Test] (Test Method) 4 ml of a plasminogen-removing fibrinogen type 2-0.6% aqueous solution was placed in a 9 cm petri dish, 4 ml of 0.1 M phosphate buffer of pH 7.4 was added, and the mixture was stirred. 0.1 ml of thrombin (10 units / ml) was added dropwise, mixed gently and left for 30 minutes. Fibrinogen was converted to fibrin by the addition of thrombin and formed a gel. 30 μl of a mixture of 0.1 ml sample and 0.1 ml plasmin solution (10 units / ml)
Was placed on the gel of a petri dish and left at 37 ° C. for 2 hours. As the sample, a 5 mM 33% dimethyl sulfoxide solution was used. Then, the dissolved area of fibrinogen was measured. Further, the same experiment was conducted using a 33% dimethylsulfoxide aqueous solution instead of the sample, and the inhibition rate of the plasmin activity was obtained by the following formula.

[0039]

[Equation 1]

When tranexamic acid and ε-aminocaproic acid were tested as positive controls, 50% inhibitory concentrations were tranexamic acid 30 mg / ml and ε-aminocaproic acid 40.
It was mg / ml. The results are shown in Table 7.

[0041]

[Table 7]

[Collagenase Inhibition Test] (Test Method) Type I Collagenase Activity Measurement Kid YU-1
Experiments were performed using 6001 Cosmo Bio. That is, fluorescence labeled type I collagen (50 μm
g / 50 μl / tube) and reagents, samples, etc. as shown in Table 8 below. Here, the neutralization solution, enzyme reaction terminator, collagenase solution, and TOTAL blank in Table 8 below
It is as follows. The neutralizing solution is a 0.1 M tris (oxymethyl) aminomethane (abbreviation Tris) -hydrochloric acid buffer solution having a pH of 7.5, 0.4 M NaCl, 0.01
It contains M CaCl ₂ and NaN ₃ (sodium azide).
The enzyme reaction terminator is an O-phenanthrone (containing ethanol) solution. The collagenase solution is a 1 unit / ml, 2-fold diluted neutralizing solution of collagenase (1000 unit / mg manufactured by Amano). The TOTAL blank is used to check the correctness of the test method when the concentration of the collagenase solution is too high.

[0043]

[Table 8]

Then, the collagenase activity inhibition rate was calculated by the following formula. The results are shown in Table 9 below.

[0045]

[Equation 2]

[0046]

[Table 9]

(Use test) The faces of 6 women were divided into left and right sides, and the lotion and cream of the example were set on one side and the lotion and cream of the comparative example were set on the other side and used once or more daily. After three months,
The whitening, the prevention of rough skin, the gloss of the skin and the suppleness of the skin were evaluated. In the comparative example, the extracts of various Terminaria horrida of Production Examples 1 and 2 from Examples 1 and 2 were replaced with water (Comparative Examples 1 and 2). In addition, 12 people were divided into 2 groups and tested using the samples shown in Table 10 below.

[0048]

[Table 10]

The evaluation is performed according to the following evaluation criteria, and the results are summarized in Table 11 below. (Evaluation Criteria) The Example is Better 3 The Example is Better 2 The Example is Better 1 No Difference 0 The Comparative Example Better -1 The Comparative Example Better- 2 The comparative example is much better -3

[0050]

[Table 11]

Results of tyrosinase activity inhibition test (Table 1), hyaluronidase activity inhibition test results (Tables 2 and 3), active oxygen inhibition test results (Table 4), antioxidant test results (Tables 5 and 6), Antiplasmin test results (Table 7), collagenase inhibition test results (Table 9), usage test (Table 1)
As is clear from 1), the cosmetic containing the solvent extract of Terminaria horida of the present invention suppresses the activity of tyrosinase, the activity of hyaluronidase and active oxygen, and the inhibition of antiplasmin and collagenase is extremely high, and whitening,
It has been found that it is effective in preventing rough skin, shining the skin, and improving the elasticity of the skin.

[0052]

INDUSTRIAL APPLICABILITY According to the present invention, not only has a strong whitening effect, but also a strong active oxygen suppressing effect, an antioxidant effect, a hyaluronidase activity inhibiting effect, an antiplasmin effect, a collagenase activity inhibiting effect, and is effective for rough skin and the like. Cosmetics are provided.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI technical display location // C12N 9/99 (72) Inventor Kenji Shimomura 3-16-32 Funae, Ise City, Mie Prefecture (72) ) Inventor Koichi Iida 56-1 Kurose-cho, Ise City, Mie Prefecture (72) Inventor Yukihisa Yamabe 3-1-6 Kawasaki, Ise City, Mie Prefecture

Claims (1)

[Claims] 1. Terminalia horida
cosmetics containing a solvent extract of orrida).