JPS63183535A - External preparation for skin - Google Patents

Abstract

PURPOSE:To obtain an external preparation for skin preventing aging of ski, providing smooth, moist and young skin free from wrinkles, containing a compound having a specific basic skeleton of guanosine 3',5'-cyclic-phosphate compound, etc. CONSTITUTION:An external preparation for skin containing a compound, preferably a compound shown by formula II (R1-R4 are H, 1-221C acyl or 1-22C alkyl; X is H, halogen, mercapto, thio-containing group, amino, aminoalkyl or OH; M is H or salt-forming cation), having a basic skeleton shown by formula I, as an active ingredient. The amount of the active ingredient contained in the external preparation for skin is 0.001-5wt.%, especially 0.01-2wt.%. When the external preparation is applied to the skin, excellent effects are shown as a result of multiplication promoting action of corium fibroblast, production promoting action of collagen elastic fiber, controlling action on melanin dyestuff metabolism, etc., by percutaneous absorption.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention prevents skin aging by activating skin cells, promoting metabolism, etc., and provides wrinkle-free, smooth, moist, and youthful skin. It relates to external skin preparations such as cosmetic creams and ointments.

[Conventional technology]

Conventionally, animal and plant extracts, vitamins, amino acids, nucleic acids, etc. extracted from natural sources have been added to cosmetics and other external skin preparations to prevent skin aging, but their effects are limited to supplementary ones. Because of this, no significant anti-aging effect on the skin could be expected even when incorporated in relatively large amounts.

On the other hand, prosufuglandin (Japanese Unexamined Patent Publication No. 1843'6, 1972), which is a physiologically active substance that exists in small amounts in living organisms,
It has been proposed to use F (epidermal growth factor) and urogastrone in cosmetics (Japanese Patent Application Laid-Open No. 61-5006). However, these methods have the following problems: (1) Since the compound is a high molecular weight substance, its permeability to cells is low and it cannot exhibit sufficient effects; (ii) EGF and urogastrone are protein-based substances; Because it is a substance, it is easily degraded by proteases within cells and cannot exert sufficient effects (iii)
On the other hand, it is not practical due to the risk of side effects and safety issues.

[Problem that the invention seeks to solve]

Therefore, an object of the present invention is to provide a practical skin preparation for external use that has an excellent anti-aging effect against skin aging phenomena such as the appearance of wrinkles, has no side effects, and is highly safe.

[Means for solving problems]

The present invention has discovered that a specific nucleic acid-related substance, which exists in trace amounts in each tissue and cell of the living body and is thought to have various regulatory functions, has an excellent effect on preventing skin aging. This was done based on the knowledge that the above-mentioned problems can be effectively solved by incorporating the compound into a skin preparation for external use.

That is, the present invention provides an external skin preparation characterized by containing a compound having a basic skeleton represented by the following formula (13).

Therefore, any compound can be used as long as it has the basic skeleton represented by the above formula (13). Preferably, guanosine 3', 5' tetra-phosphoric acid represented by the following general formula [■] (in the formula [■], R1, R2,
A compound in which R3, R4, x, and M are all hydrogen; hereinafter cG
It is abbreviated as MP. ), its specific derivatives and its salts can be used.

(In formula [■], R1, R2, R3, and R4 are hydrogen, an acyl group having 1 to 22 carbon atoms, or an alkyl group having 1 to 22 carbon atoms, and X is hydrogen, halogen, mercapto group, thio containing group, amine group, aminoalkyl group, or hydroxyl group, and M represents hydrogen or a salt-forming cation.) Here, in the formula CID, R,, R2, R3, and R4 are hydrogen, carbon number 1- 22, particularly preferably an acyl group having 1 to 12 carbon atoms, and an alkyl group having 1 to 22 carbon atoms, particularly preferably 1 to 12 carbon atoms.

In addition, as X, hydrogen, halogen such as bromine, iodine, chlorine, fluorine, mercapto group, thioalkyl group having 1 to 4 carbon atoms, thio-containing group such as 4-chlorophenylthio group, thiobenzyl group, amino group, carbon number 1 to 4 aminoalkyl groups and hydroxyl groups are preferred, and M is preferably hydrogen, alkali metals such as sodium, potassium, and lithium, and tris(hydroxymethyl)aminomethane.

Incidentally, the acyl group and the alkyl group may have a substituent such as a halogen, and may contain an aromatic ring. Alternatively, it may be an acyl group derived from a dibasic acid.

cGMP used in the present invention is a substance whose importance has recently been recognized, as it exists in trace amounts in living organisms and has the ability to regulate various hormone actions. Currently, cGMP and various derivatives produced by fermentation or synthesis methods are commercially available as biochemical reagents.

Specifically, the compound represented by the general formula [■] used in the present invention includes one or a mixture of two or more of the compounds shown in Table 1 and their sodium salts, potassium salts, lithium salts, etc. Illustrated.

In Table 1, ClPh5 represents a 4-chlorophenylthio group (the same applies hereinafter).

The above-mentioned nucleic acid-related substances used in the present invention are usually contained in various skin preparations ranging from 0,001 to
5% by weight (hereinafter abbreviated as %), preferably 0.0
The content is preferably 1 to 2%.

In addition to the above-mentioned essential ingredients, the skin external preparation of the present invention contains oil,
Raw materials commonly used in cosmetics such as water, surfactants, humectants, lower alcohols, thickeners, antioxidants, chelating agents, pH adjusters, preservatives, fragrances, and pigments can be blended. Specifically, oils include oils and fats such as olive oil, jojoba oil, and hydrogenated oil; waxes such as spermaceti wax, beeswax, and lanolin;
Examples include hydrocarbons such as liquid paraffin, ceresin, and squalane, fatty acids such as stearic acid and oleic acid, alcohols such as cetanol, stearyl alcohol, lanolin alcohol, and hexyldecanol, and esters such as isopropyl myristate and butyl stearate. be done. Examples of surfactants include anionic surfactants such as sodium stearate, sodium cetyl sulfate, polyoxyethylene lauryl ether phosphate, and sodium N-acylglutamate, and cationic surfactants such as stearyldimethylbenzylammonium chloride and stearyltrimethylammonium chloride. Active agent, alkylaminoethylglycine hydrochloride solution, amphoteric surfactant such as lecithin, glyceryl monostearate, sorbitan monostearate,
Sucrose fatty acid estenopemonostearate propylene glycol, polyoxyethylene oleyl ether, monostearate polyethylene glycol, monopalmitate polyoxyethylene sorbitan, polyoxyethylene coconut fatty acid monoethanolamide, polyoxyethylene polyoxypropylene glycol, poly Examples include nonionic surfactants such as oxyethylene castor oil and polyoxyethylene lanolin. Furthermore, as humectants, glycerin, 1,3-butylene glycopropylene glycol, etc. are used, as lower alcohols, ethanol, inpropatol, etc. are used, and as thickeners, polyethylene glycol, carboxymethyl cellulose sodium, etc. are used. As antioxidants, dibutylhydroxytoluene, butylhydroxyanisole, propyl gallate, etc., as chelating agents, edetate disodium, ethane hydroxy diphosphate, etc., as pH adjusters, citric acid, sodium citrate, etc. , boric acid, borax, sodium hydrogen phosphate, etc., as preservatives,
Examples include metinopeparaoxybenzoic acid, ethinobedehydroacetic acid, salicylic acid, and benzoic acid. Note that the optional components are not limited to these. By appropriately blending the above essential ingredients and optional ingredients, for example, 0.001 to 5% of the essential ingredients
, oil content 0-80% as optional ingredients: surfactant 0.5-80%
It is possible to provide a skin external preparation containing 12%, 2-15% humectant, 11-95% purified water, and a trace amount of preservative.

Specifically, it can be used in various product forms such as lotion, cream, pack, lotion, skin milk, emulsion, and ointment.

To give a specific example, the lotion contains: 0.01-2% of the essential ingredients of the present invention, 2-2% of lower alcohol.
10%, surfactant 0.5-1%, humectant 3-7%, p
11 regulator 0.05-0.2%, purified water 80-95%,
A composition containing a trace amount of preservative, a trace amount of pigment, and a trace amount of fragrance, as a skin cream: essential ingredients 0.01-2%, oil 20-70%, surfactant 2-7%, humectant 5-10% , purified water 11-73%,
A composition containing trace amounts of preservatives and trace amounts of fragrance, as a lotion: essential ingredients 0.01-2%, lower alcohol 5-10%,
Surfactant 0.5-2%, humectant 2-8%, antioxidant 0.01-0.05%, chelating agent 0.02-0.1%
, pH adjuster 0.1-1%, purified water 77-92%, a trace amount of preservative, a composition containing a trace amount of fragrance, skin milk: essential ingredients 0.01-2%, oil content 20-40%, A composition containing 2-5% surfactant, 53-78% purified water, trace amounts of preservatives, and trace amounts of fragrance, as an emulsion: n Essential ingredients 0.01-2%, oil content 10-30%, surfactant 1-5%, moisturizer 5-10%, purified water 53-84%,
Compositions and ointments containing trace amounts of preservatives and fragrances include: 0.01-2% essential ingredients, 40-60% oil, 1-12% surfactant, 8-15% humectant, 12-12% purified water. 51%
, a composition containing trace amounts of preservatives (parent ointment), essential ingredients 0.01-2%, oil content 95-99%, purified water 0
Examples include compositions (oil-based ointments) containing ~5%.

The details of how the topical skin preparation of the present invention exhibits an excellent anti-aging effect against skin aging phenomena such as the appearance of wrinkles are unknown, but first of all, the main cause of skin aging is the skin tissue. Rather than the state of the epidermis on the surface of the skin, it is assumed that this is due to the fact that the fibrous proteins collagen and elastin that make up the dermis decrease with aging, and their structure changes and becomes brittle. Therefore, based on the experiments described below, we extensively searched for substances that promote the proliferation of fibroblasts, which are cells that produce and secrete collagen and elastin in the dermis. It has been revealed that it has promoting and fibrosis-promoting effects. Furthermore, it has been revealed that it has the effect of inhibiting the production and deposition of melanin pigment, and as shown in the examples, it is also effective in preventing skin aging such as wrinkles in the human body.
The improvement effect was confirmed.

Therefore, when the cosmetic is applied to the skin, the above-mentioned excellent effects are exerted as a result of transdermal absorption, promoting the proliferation of dermal fibroblasts, promoting the production of collagen and elastin fibers, and regulating melanin pigment metabolism. It is estimated that

The above-mentioned nucleic acid-related substances used in the present invention originally exist widely in living organisms and are considered to be extremely safe.
No practical problems were observed in terms of skin irritation, skin sensitization, etc., and it was confirmed to be highly safe.

〔Effect of the invention〕

According to the present invention, by activating skin cells and preventing skin aging, it is possible to prevent the appearance of wrinkles and give smooth, moist, youthful skin, which is much more effective than conventional products. Provided are skin preparations for external use, such as cosmetics, which have the same properties and are extremely safe. Furthermore, the external skin preparation of the present invention has the excellent effect of preventing the occurrence of bleeds.

Therefore, the skin external preparation of the present invention includes various cosmetic creams,
Lotion, lotion, skin milk, emulsion, pack agent,
It can be widely used in various forms such as ointments.

Next, the present invention will be explained using test examples and examples.

[Test example]

Test Example 1 The fibroblast proliferation promoting effect of the nucleic acid-related substance used in the present invention was evaluated as follows.

Human dermis-derived fibroblasts were cultured in medium with and without the above compound under serum-suppressed conditions, and the effect on proliferation was investigated.

That is, the cells were cultured in a C02 incubator at 37 degrees for 24 hours in a medium (pH 7, 6) containing Eagle's MEM medium supplemented with 10% fetal bovine serum, etc., and then replaced with a medium containing suppressed serum to 1%. The cells were then cultured for an additional 24 hours. Here, the medium was replaced with a medium containing 10 −5 M of the test substance and 1% serum, and culture was continued.

Thereafter, the medium was replaced every other day. Table 2 of the results
Shown below. Here, the number of cells after 14 days in the case where no test substance was added (control) was set as 100, and the cell number when the test substance was added was expressed as a relative value.

In Table 2, the following abbreviations are used (the same applies below).

MB; Monobutyryl MS; Monosuccinyl 1Phs; 4-chlorophenylthio DB; Dibutyryl DS; Disuccinyl Further, the positions of the substituents of the cGMP derivative are shown as follows.

N2; bonding position when R2 or R3 is other than hydrogen.

○”; Bonding position when R4 is other than hydrogen.

8; Bonding position when x is other than hydrogen.

Table 2 From the results in Table 2, it can be seen that all of the active ingredients of the present invention exhibit significant growth-promoting effects on human fibroblasts.

Fibroblasts present in the dermis of the skin are cells that produce structural proteins such as collagen and elastin, which are responsible for maintaining skin flexibility and elasticity. Therefore, promoting the proliferation of fibroblasts is a necessary condition for promoting the production of collagen and elastin, and as a result, improving the softness and elasticity of the skin.

Test Example 2 The collagen production promoting effect of the nucleic acid-related substance used in the present invention was evaluated as follows.

One group consisted of 6 male Wistar rats aged 6 weeks. After hair was removed from the backs of the rats, the skin was cut into a circular shape (about 15 mm in diameter) and treated with 10% chloride containing 50 mM of the above compound.
0.1 ml each of ethanol solution or 10% ethanol solution (control) was applied twice daily for 6 consecutive days. After layer killing on the 7th day, the surgical site was excised.
The weight of granulation, which is the newly formed dermis, and the amount of hydroxyproline, which is a constituent amino acid unique to collagen, were measured as indicators of the amount of regenerated collagen. Display the results -
Shown in 3. Here, the granulation weight and hydroxyproline amount of the control were each set as 100, and relative values are shown.

From the results in Table 3, it was found that the active ingredients of the present invention clearly increased the weight of regenerated granulation and the amount of hydroxyproline, indicating that the amount of collagen in the dermis increased. There is.

In other words, as a result of the promotion of proliferation of fibroblasts present in the skin dermis, the weight of the dermis increases, and at the same time, collagen production is promoted, preventing skin aging such as a decrease in elasticity and the appearance of fine wrinkles. This shows that it is possible to improve.

Test Example 3 The effect of the active ingredient of the present invention on the strength of collagen fibers and lastin fibers in the dermis was evaluated as follows.

A group of eight 6-week-old Wistar male rats were made. After removing hair from the backs of the rats, a 4 cm sharp linear wound was made along the dorsal midline with a scalpel, and 3
The area was sutured using a Mitsuhel needle. The Mitsuhel needle was removed on the fourth postoperative day. Physiological saline containing 5 QmM of the test substance or physiological saline containing no test substance (control) at 0.1 mA each was administered to the incision site once a day for 7 consecutive days.

After layer killing on the 8th day, the skin at the incision site is cut out, three skin pieces with a width of 1 cm perpendicular to the incision line are prepared for each individual, and the skin pieces at the incision site are cut using a tensile strength tester. Tensile strength
g/Cm) was measured and used as an index of the strength of regenerated collagen/elastin fibers.

The results are shown in Table 4. Here, the tensile strength of the control was set as 100 and expressed as a relative value.

Table 4 In this test, if the relative value is 110 or more,
It is generally judged that the tensile strength has been improved, but Table 4
From the results, a clear increase in the tensile strength of regenerated collagen and elastin fibers was observed for all active ingredients of the present invention.

In other words, this indicates that the elasticity and flexibility of the skin may be improved by increasing the tensile strength of the regenerated collagen and elastin fibers.

〔Example〕

Example 1 Components 1 to 6 and components 7 to 10 shown in Table-5 were separately prepared at 80%
After heating and dissolving at ℃, the mixture was mixed and emulsified, and while cooling, component 11 was added and uniformly dispersed to prepare the cream shown in Table 5.

In addition, the numerical values in the table are weight % (the same applies hereinafter).

Melanin pigment production and
The influence on deposition was evaluated as follows.

After hair removal, a cream containing 0.5% of the active ingredient and a cream containing no active ingredient were applied to the backs of colored guinea pigs (3 animals per group) twice a day in the morning and evening for 14 consecutive days. The difference in skin tone before and after application was measured using a color difference meter. The results are shown in Table-6.

Table 6 The numerical values in the table indicate the L value (lightness), and the larger the value, the whiter the skin.

As is clear from the results in Table 6, the cream containing cGMP and its derivatives has a superior decolorizing effect compared to the control.

That is, it is shown that the active ingredient of the present invention has an effect of suppressing the production and deposition of melanin pigment.

Example 2 A homogeneous mixture of components 3 and 7 to 9 shown in Table 7 was added to an aqueous solution in which the other components were dissolved, and the mixture was solubilized.
The lotion shown in was prepared.

Table 7 Next, the effectiveness of the lotion thus prepared was evaluated as follows.

Twice a day (morning and evening) to 20 women (30 to 50 years old)
The products of the present invention and the comparative example were used separately on the left and right sides of the face using the half-face method for three consecutive months, and then the degree of improvement in skin elasticity, skin luster, rough skin, and fine wrinkles was examined. The results are summarized in Table-8.

From the results in Table 8, N2. O”-DBcGMP ・N
The lotion of the present invention containing A showed excellent effects in improving skin elasticity, luster, rough skin, and fine wrinkles, compared to the comparative example without additives.

From the above results, N2. It is clear that ◯''-DBcGMP/Na has excellent effects on skin activation and anti-aging.

In addition, no abnormality was observed in the condition of the skin during and after using the above lotion for three months.

In addition, the N2. used in the lotion of the present invention. Q2'-DBcG
The results of the safety test of MP-Na are shown below.

Acute oral toxicity (rat): LDso value; 5 g/kg or more Skin irritation (1) - Secondary irritation (guinea pig) = 5%; No irritation (2)
Phototoxicity (guinea pig): Negative (3) Eye mucosal irritation (
Rabbit) = 5%; Non-irritating skin sensitization (1) Sensitization (guinea pig): Negative (2) Photosensitivity (guinea pig): Negative Human batch test: 5%; Based on the results of no irritation or higher, N2 ,0''-DBcGMP・Na was confirmed to have extremely high safety.

Example 3 Components 1 to 5 and components 6 to 10 shown in Table-9 were separately prepared at 80%
After heating and dissolving at ℃, the two were mixed and emulsified, and while cooling,
Component 11 was added and uniformly dispersed to prepare the emulsion shown in Table 9.

Table 9 The effectiveness of the emulsion thus prepared was examined in the same manner as in Example 2 by having 20 women use it.

The results are summarized in Table-10.

From the results in Table 10, the emulsion of the present invention containing CGMP-Na showed superior effects in each evaluation item compared to the comparative example without additives. In addition, no abnormality was observed in the condition of the skin during and after using the above emulsion for three months.

When the safety of cGMP-Na was tested using the same method as in Example 2, no problems such as skin irritation were observed, and it was confirmed that it was extremely safe.

Example 4 N2. O”-DBcGMP-Na instead of N2-M
B c GMP -Na, O” -MB c GM
P・Na, N2. When the anti-aging effect on the skin was investigated in the same manner as in Example 2 except that O''-DScGMP-Na or 8-Br cGMP-Na was used, excellent effects similar to those in Example 2 were obtained.

Example 5 An ointment with the composition shown in Table 11 was prepared and applied to the pigmented areas of six men and women with pigmented spots (spots and freckles) twice a day in the morning and evening.
The ointment was applied for a month. The brightness of the pigment spots before and after application was measured using a color difference meter. The results are shown in Table-12.

Table-11 Table-12 The numerical values before and after the test in the table indicate the L value (lightness).

As is clear from the results in Table 12, N2. O”-DB
It can be seen that the brightness of pigment spots in the ointment containing cGMP-Na or cGMP-Na is increased compared to the ointment containing no cGMP-Na (comparative example). That is, it can be seen that cGMP and its derivatives, which are the active ingredients of the present invention, have an excellent effect on improving pigment spots (spots and freckles) in addition to the effect of improving wrinkles. In addition, no abnormality was observed in the condition of the skin during and after using the ointment for three months.

Example 6 N2. used in Example 5. O”-DBcGMP・NaS
N′-MBcGMP-Na instead of cGMP-Na
, O”-MBcGMP-Na or 8-BrcGMP
A cream containing -Na was prepared and evaluated in the same manner as in Example 5. As a result, almost the same pigmentation improvement effect as in Example 5 was obtained.

Claims (2)

[Claims] (1) An external skin preparation characterized by containing a compound having a basic skeleton represented by the following formula [I]. ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・〔I〕 (2) The compound having the basic skeleton represented by the formula [I] is a guanosine 3',5' represented by the following general formula [II]
- The skin external preparation according to claim (1), which is a cyclic phosphoric acid compound. ▲There are mathematical formulas, chemical formulas, tables, etc.▼... [II] (In the formula [II], R_1, R_2, R_3, R_4 are hydrogen, an acyl group having 1 to 22 carbon atoms, or an alkyl group having 1 to 22 carbon atoms. group, X is hydrogen, halogen, mercapto group, thio-containing group, amino group, aminoalkyl group, or hydroxyl group, and M represents hydrogen or a salt-forming cation.)