JPH11279042A - Skin preparation for external use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a skin preparation for external use capable of manifesting the same preventing actions on the formation of wrinkcles on skin or suppressing actions on keratinization of the skin as those of a retinoid by a single agent by making the preparation include an oligosulfated hyaluronic acid as an active ingredient therein. SOLUTION: This skin preparation for external use contains an oligosulfated hyaluronic acid containing, e.g. a recurring unit represented by the formula (R<1> to R<4> are each H or SO3 H, with the proviso that 10-90% of R<1> to R<4> based on the total number thereof occupy SO3 H; (n) is an average value so as to provide the viscosity-average molecular weight of 1,000-50,000) as an active ingredient. The oligosulfated hyaluronic acid which is the active ingredient can be obtained by treating a high-molecular weight oligohyaluronic acid prepared from various kinds of natural sources with, e.g. an enzyme (hyaluronidase) and then sulfating the resultant treated substance according to a known sulfating method. Thereby, the resultant preparation for external use is capable of suppressing the keratinization of skin and having suppressing actions on secretin intervening properties in combination and maintaining a healthy skin state.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to the technical fields of cosmetics and dermatology, and more specifically, to provide new uses of oligosulfated hyaluronic acid in the above fields.

[0002]

2. Description of the Related Art β-DN-acetylglucosamine and β-DN-acetylglucosamine
Hyaluronic acid, which is a linear high-molecular polysaccharide formed by alternately bonding -D-glucuronic acid, is relatively easily available among mucopolysaccharides and exhibits specific physicochemical properties. Itself or various derivatives thereof are used for medical or cosmetic purposes. A wide variety of new proposals have also been made. The present inventors have also found that, for example, a sulfated hyaluronic acid, which is a derivative of hyaluronic acid, exhibits a strong anti-inflammatory effect mediated by selectin, which is one of the adhesion factors. (See WO 96/24362).

On the other hand, regarding the use of mucopolysaccharide in the technical field of cosmetics, Japanese Patent Publication No. 7-121852 discloses mucopolysaccharide and / or a derivative thereof, vitamin E and / or a derivative thereof, and a physiological salt solution. It describes that a cosmetic containing the compound prevents skin dryness and rough skin. Hyaluronic acid, hyaluronic acid as mucopolysaccharide used
Protein complexes, chondroitin tetrasulfate, and the like. In addition, Japanese Patent Publication No. 4-65047 discloses a low-molecular-weight acidic mucopolysaccharide, for example, an average molecular weight of 1,000 to 900.
0 hyaluronic acid or average molecular weight 2000-900
It has been suggested that a combination of chondroitin sulfate 0 and γ-aminobutyric acid has a skin aging preventing effect and the like.

[0004]

SUMMARY OF THE INVENTION Hyaluronic acid itself or low-molecular-weight hyaluronic acid and other acidic mucopolysaccharides themselves have been conventionally used in cosmetics in the hope that a skin moisturizing effect will be brought about. Has been formulated.
However, as described in both publications,
These acidic mucopolysaccharides alone do not show a significantly superior stratum corneum turnover effect, and only when combined with certain other drugs, only the significant effect described above is observed. Certainly, the usefulness of the combination with these specific drugs is recognized. Nevertheless, in order to apply to a wide variety of forms of cosmetics, it is necessary to have a drug that does not have the specific combination as described above, that is, a single component that shows some significant effect.

[0005] By the way, in human skin, epidermal atrophy and accumulation of keratinocyte layer increase remarkably with aging, and wrinkles (so-called
It is thought to contribute to the formation of aging skin). As a single agent most effective in preventing such wrinkle formation, a retinoid is currently used.

[0006] That is, an object of the present invention is to provide an active ingredient which, as a single agent, exhibits almost the same anti-wrinkle formation or skin keratinization effect as a retinoid.

[0007]

As described above, the present inventors have shown that high-molecular-weight sulfated hyaluronic acid has a strong anti-inflammatory effect mediated by selectin. Such sulfated hyaluronic acid has been found to have a slightly reduced anti-inflammatory effect with a decrease in molecular weight, but, on the contrary, a markedly improved skin keratinization inhibitory effect. Further, as far as the present inventor knows, such low molecular weight (so-called oligo) sulfated hyaluronic acid has not been published in known literature.

Therefore, according to the present invention, there is provided an external preparation for skin containing oligosulfated hyaluronic acid as an active ingredient. Although not limited, the details will be described later, but in consideration of the action and effect of oligosulfated hyaluronic acid found by the present inventors, the above-mentioned external use effect on the skin is particularly aimed at suppressing keratinization of the skin. It is provided as an external preparation for skin.

[0009]

DETAILED DESCRIPTION OF THE INVENTION The oligosulfated hyaluronic acid used in the present invention is not limited as long as it meets the purpose of the present invention.
The degree of sulfation is not limited. However,
More specific examples of oligosulfated hyaluronic acid include compounds of the formula (I)

[0010]

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(Wherein R ¹ , R ² , R ³ and R ⁴ each independently represent a hydrogen atom or an SO ₃ H group, and n is such that the viscosity average molecular weight is from 1,000 to 50,000). Any one of the average values, and R ¹ , R ² , R ³ and R ⁴
10-90% per total number of SO ₃ H groups occupied) oligo sulfated hyaluronic acid comprising recurring units or a salt thereof represented by, and the like.

Although not limited, when the above oligosulfated hyaluronic acid is used for the purpose of the present invention,
The above-mentioned n number is preferably a value having a viscosity average molecular weight of 1,000 to 10,000, and more preferably 2,000 to 7.0.
Those having a value of 00 are more preferred. This is because they show appropriate skin permeability and also have a certain moisturizing effect. When the skin permeability is mainly desired, it is preferable to take the value of n so that the viscosity average molecular weight becomes 5,000 or less.

The number n of repeating units according to formula (I) is SO ₃
The range varies depending on the degree of substitution of the H group, that is, the sulfation rate, but typically, about 6 to 10 can be mentioned. Such a repeating unit having a sulfation ratio of 20 to
50% is particularly preferred for the purposes of the present invention.

The oligosulfated hyaluronic acid is
An alkali metal or alkaline earth metal, preferably
It can also be used in the form of salts with sodium, potassium, lithium or salts with ammonia or organic amines.

The above-mentioned sulfated oligohyaluronic acid or a salt thereof according to the present invention can be produced from the corresponding oligohyaluronic acid according to a per se known sulfation method. The corresponding salt can be produced by subjecting it to a salt forming method using a metal hydroxide or carbonate or ammonia or an organic amine.

Oligohyaluronic acid is obtained by treating high molecular weight (for example, 2,000 kDa) obtained from various natural sources with high shear stress or by treating with an enzyme (hyaluronidase) (for example, A. Sattor et al.). , J. Invest. Der
matol. Vol. 103: 576-579, 1994). According to the enzyme treatment, oligohyaluronic acid having a narrow molecular weight distribution can be advantageously obtained by selecting the enzyme activity and reaction time to be used. In the average molecular weight range according to the present invention,
An almost monodisperse oligohyaluronic acid can also be obtained through a process such as gel filtration or fractional precipitation with an organic solvent such as acetone.

The oligohyaluronic acid thus obtained is
The oligosulfated hyaluronic acid of the present invention can be converted by a sulfation method known per se. In general, it is advantageous to use a sulfuric acid-trimethylamine complex as the sulfating agent. The ratio of oligohyaluronic acid to the sulfating agent used can be arbitrarily selected depending on the desired sulfation rate. The reaction conditions include a temperature of 50 to 60 in an aprotic polar solvent (for example, dimethylformamide).
Conditions can range from tens of hours to days at ° C.

The resulting oligosulfated hyaluronic acid can be purified according to a method known per se in the purification of oligosaccharides, if necessary, through gel filtration or fractional precipitation with acetone. The salt formation reaction may be performed before or after the above purification treatment.

The external preparation for skin of the present invention containing oligosulfated hyaluronic acid as described above has a main function of suppressing the keratinization of the skin. Although it is low, it also has a selectin-mediated effect of inhibiting the infiltration of inflammatory lymphocytes into skin tissue, and is also effective against dullness, atopic skin, contact and irritable dermatitis, sensitive skin and dry skin. Have.

The content of oligosulfated hyaluronic acid in the external preparation for skin of the present invention is 0.0001 to 30.0% by weight, preferably 0.001 to 30.0% by weight, based on the total weight of the external preparation.
15.0% by weight. If it is less than 0.0001%, the effects of the present invention cannot be sufficiently exhibited, and if it exceeds 30.0% by weight, it is difficult to formulate the composition, which is not preferable. Also, 1
Even if the content is 5.0% by weight or more, no significant improvement in the effect is observed.

In addition to the oligosulfated hyaluronic acid as an active ingredient, the skin external preparation of the present invention contains components usually used in skin external preparations such as cosmetics and pharmaceuticals, for example, other whitening agents, moisturizing agents, oxidizing agents, and the like. Inhibitors, oily components, ultraviolet absorbers, surfactants, thickeners, alcoholic agents, powder components, coloring agents, aqueous components, water, various nutrients, and the like can be appropriately compounded as necessary.

Other sequestering agents such as disodium edetate, trisodium edetate, ethylenediaminehydroxyethyl-trisodium triacetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, caffeine, tannin, verapamil , Tranexamic acid and its derivatives, licorice extract, glabridine,
Hot water extract of fire thorn fruit, various crude drugs, drugs such as tocopherol acetate, glutyrrhizic acid and its derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid and other whitening agents Ingredients, sugars such as glucose, fructose, mannose, sucrose, trehalose and the like can also be appropriately compounded.

The external preparation for skin of the present invention may be any of those used in, for example, ointments, creams, emulsions, lotions, packs, bath preparations, and conventional external preparations for skin, and the dosage form is not particularly limited.

[0024]

Next, the present invention will be described in more detail by way of examples. Note that the present invention is not limited by this.

Evaluation of local stratification control action using PAM212 cells This evaluation test is carried out by comparison with a retinoid (control), which has been widely confirmed to have an excellent epidermal keratinization inhibitory action. It is possible to evaluate the degree of keratinization based on a change caused by a change in calcium concentration, such as an increase in keratinization in normal epidermal cells, which depends on calcium concentration. As a result, PAM212 cells can use local stratification that occurs in a calcium ion-dependent manner as an indicator. That is, in the following experiments, local stratification occurring under high calcium concentration was defined as a positive control in which the local stratification was significantly suppressed by the addition of retinol, and oligosulfated hyaluronic acid (20% of the total hydroxyl groups) was used. Sulfation rate).

For comparison, oligohyaluronic acid (unsulfurized oxide) having the same molecular weight was used.

In addition, PAM212 cells are BALB / C
This is the lower part of the cells established from mouse keratinocytes.

(Test method) 4 × 10 4 PAM212 cells
^The cells were adjusted to ⁴ cells / well and seeded in a 24-well. (80 × 10 ⁴ cells / mL cell suspension was added with a 2 mL pipette, and a drop (50 μL) was added. Further, 0.5 mL of medium was added with a 10 mL pipette. did.
The four sides of the 24-well were patted lightly about 10 times. ). The medium was EMEM / 10% untreated serum.

The cells were cultured for 2 days in static culture, and when they became 80-90% confluent, a drug was added and the effect was compared with that of retinol. Since the drug was adjusted to a 2% (w / v) aqueous solution, dimethyl sulfoxide (D
MSX), dilute 10-fold, 2 × 10 ⁻¹ , 2 × 10 ⁻² , 2
A 10% ^-3 % DMSO solution (DMSO solution of drug) was prepared. 5 μL of DMSO solution of drug per 1 mL of medium
At a final concentration of 10 ⁻³ , 10 ⁻⁴ , and 10 ^−5.
%. Similarly, DMSO, retinol (final concentration 10
^-7 M) was added. The medium is EMEM / 5% defatted serum.

The number of local stratifications occurring 24 hours after drug addition was measured and compared with DMSO and retinol. DMS
With the number of local stratifications in O being 100%, the retinol inhibition rate (%) was determined. 80 to 1 against retinol suppression
It was determined that 00% was significant, 60-80% was valid, 40-60% was undeterminable, and less than 40% was invalid.

In order to confirm that this effect is the effect of the test drug, the number of cells was replaced with the amount of DNA and measured.
The presence or absence of cytotoxicity was determined. An agent which did not show cytotoxicity and suppressed local stratification was determined to be an effective substance.

(Test Results) The effect of oligohyaluronic acid on local stratification was examined. Control group 100.1 ±
Oligohyaluronic acid was 81.1 ± 14.1% at a concentration of 10 ⁻³ % compared to 10.6%. This drug had no local stratification inhibitory effect.

The effect of oligosulfated hyaluronic acid (from Example 2) on local stratification was examined. Control group 99.
Oligosulfated hyaluronic acid is 9 ± 14.7%
It showed 86.5 ± 23.1% at a concentration of 10 ⁻⁴ %, and showed a significant difference (unpaired t-test) from the control group, indicating that this drug has a local stratification inhibitory effect. Was. Retinol is 9.5 ± 2.1 at 10 ⁻⁷ M
%) As described above, the oligosulfated hyaluronic acid of the present invention, which has excellent skin permeability and suppresses local stratification inhibitory action, is useful, for example, as an active ingredient of cosmetics. Examples of the formulation of the skin external preparation of the present invention in various dosage forms are described below as examples.

Example 1 Cream (Formulation) 5.0% by weight of stearic acid behenyl alcohol 4.0 Isopropyl myristate 18.0 Self-emulsifying glyceryl monostearate 3.0 1,3-butylene glycol Glycerin monostearate 10.0 Oligosulfated hyaluronic acid 0.1 α-olefin oligomer Potassium hydroxide 0.2 Sodium bisulfite 0.01 Preservatives Appropriate amount Fragrance Appropriate amount Ion-exchanged water residue (Production method) 1,3-butylene glycol and oligosulfated hyaluronic acid in ion-exchanged water The acid and potassium hydroxide are added and dissolved, and the mixture is heated and maintained at 70 ° C. (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is gradually added to the water phase, and after the addition is completed, the temperature is maintained for a while to cause a reaction. Thereafter, the mixture is uniformly emulsified with a homomixer and cooled to 30 ° C. while stirring well.

Example 2 Cream (Prescription) Stearic acid 2.0% by weight Stearyl alcohol 7.0 Hydrogenated lanolin 2.0 α-olefin oligomer 5.0 2-octyldodecyl alcohol 6.0 Polyoxyethylene (25 mol) Cetyl alcohol ether 3.0 Self-emulsifying glyceryl monostearate 2.0 1,3 Butylene glycol 5.0 Oligosulfated hyaluronic acid 0.2 Sodium bisulfite 0.03 Preservative 0.3 Perfume Appropriate amount Ion-exchanged water Production method) Add 1,3-butylene glycol and oligosulfated hyaluronic acid to ion-exchanged water, dissolve and heat to 70 ° C.
(Aqueous phase). Mix other ingredients, heat and melt at 70 ° C
(Oil phase). Add oil phase to water phase, pre-emulsify, homogenize with homomixer, and mix well
Cool to 0 ° C.

Example 3 Cream (Formulation) Paraffin 5.0% by weight Beeswax 10.0 Vaseline 41.0 Self-emulsifying glyceryl monostearate 2.0 Polyoxyethylene (20 mol) sorbitan monolaurate 2.0 Soap powder 0.1 Borax 0.2 Oligosulfated hyaluronic acid 0.1 Sodium bisulfite 0.03 Preservatives Appropriate amount Perfume Appropriate amount Ion-exchanged water residue (Production method) Add soap powder, borax and oligosulfated hyaluronic acid to ion-exchanged water. Heat and maintain at 70 ° C (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is gradually added to the aqueous phase while stirring to carry out the reaction. After completion of the reaction, the mixture is uniformly emulsified with a homomixer, and cooled to 30 ° C. while stirring well after emulsification.

Example 4 Emulsion (Formulation) 2.5% by weight of stearic acid Cetanol 1.5 Vaseline 5.0 α-olefin oligomer 10.0 Polyoxyethylene (10 mol) monooleate 2.0 Polyethylene glycol 1500 3 0.0 Potassium hydroxide 1.14 Carboxyvinyl polymer 0.05 Oligosulfated hyaluronic acid 0.05 Sodium bisulfite 0.01 Preservatives Appropriate amount Fragrance Appropriate amount Ion-exchange water residue (Preparation method) Carboxyvinyl polymer in a small amount of ion-exchange water Dissolves (phase A). Polyethylene glycol 1500 and oligosulfated hyaluronic acid are added to the remaining ion-exchanged water and kept at 70 ° C. (aqueous phase). The other components are mixed, dissolved by heating and kept at 70 ° C. (oil phase). After neutralization by adding potassium hydroxide, the oil phase is added to the aqueous phase to carry out preliminary emulsification, the phase A is added, and the mixture is uniformly emulsified with a homomixer.
Cool to ° C. Example 5 Emulsion (Formulation) Microcrystalline wax 1.0% by weight Dense wax 2.0 Lanolin 20.0 α-olefin oligomer 10.0 Squalane 5.0 Sorbitan sesquioleate 4.0 Polyoxyethylene (20 mol) Sorbitan monooleate 1.0 1.0 Butylene glycol 7.0 Oligosulfated hyaluronic acid 10.0 Sodium bisulfite 0.01 Preservatives Appropriate amount Fragrance Appropriate amount Ion-exchanged water Residue (Production method) 1,3-butylene in ion-exchanged water Glycol is added and heated to 70 ° C. (aqueous phase). The other components are mixed, heated and melted and kept at 70 ° C. (oil phase). While stirring the oil phase, the aqueous phase is gradually added thereto, and the mixture is uniformly emulsified with a homomixer. After emulsification, cool to 30 ° C with good stirring.

Example 6 Jelly (Prescription) Government-designated denatured alcohol 10.0% by weight Dipropylene glycol 5.0 Polyoxyethylene (50 mol) oleyl alcohol ether 2.0 Carboxyvinyl polymer 1.0 Sodium hydroxide 0.15 L-arginine 0.1 oligosulfated hyaluronic acid 6.0 sodium hydroxymethoxybenzophenonesulfonate 0.05 trisodium edetate 0.05 preservatives qs perfume qs Ion exchange water residue Oligosulfated hyaluronic acid is uniformly dissolved, while the government-designated denatured alcohol polyoxyethylene (50 mol)
Dissolve the oleyl alcohol ether and add to the aqueous phase. Next, after adding other components, the mixture is neutralized with sodium hydroxide and L-arginine to increase the viscosity.

Example 7 Essence (Formulation) (A phase) Denatured alcohol prescribed by the government 10.0% by weight Polyoxyethylene (20 mol) octyldodecanol 1.0 Pantothenyl ethyl ether 0.1 Oligosulfated hyaluronic acid 1 .5 Preservatives Appropriate amount (B phase) Caustic potassium 0.1 (C phase) Glycerin 5.0 1,3-butylene glycol 10.0 Sodium bisulfite 0.03 Carboxyvinyl polymer 0.2 Ion-exchanged water Residue (Preparation method) Dissolve A phase and C phase uniformly, and add A
Add phases and solubilize. Next, after adding the phase B, filling is performed.

Example 8 Pack (Formulation) (Phase A) Dipropylene glycol 5.0% by weight Polyoxyethylene (60 mol) hydrogenated castor oil 5.0 (Phase B) Oligosulfated hyaluronic acid 0.01 α-olefin oligomer 5.0 d-δ-tocopherol 0.2 Preservatives Appropriate amount Fragrance Appropriate amount (C phase) Sodium bisulfite 0.03 Polyvinyl alcohol 13.0 Government-designated denatured alcohol 7.0 Ion-exchanged water Residue (Production method) A phase, B phase , C phase are uniformly dissolved,
Add phase B to phase and solubilize. Next, this is added to the C phase and then filled.

Example 9 Solid foundation (formulation) Talc 43.1% by weight Kaolin 15.0 Sericite 10.0 Zinc white 7.0 Titanium dioxide 3.8 Yellow iron oxide 2.9 Black iron oxide 0.2 Bengala 0 0.3 Squalane 8.0 Isostearic acid 4.0 POE sorbitan monooleate 3.0 Isocetyl octoate 2.0 Oligosulfated hyaluronic acid 1.0 Preservatives Appropriate perfume Appropriate amount (Production method) Mix well with a blender, add the oily components of squalane to isocetyl octanoate, oligosulfated hyaluronic acid, preservatives, and flavors, knead well, fill into a container, and mold.

Example 10 Emulsion type foundation (cream type) (Prescription) (Powder part) Titanium dioxide 10.3% by weight Sericite 5.4 Kaolin 3.0 Yellow iron oxide 0.8 Bengala 0.3 Black iron oxide 0.2 (oil phase) decamethylcyclopentasiloxane 11.5 α-olefin oligomer 4.5 methylpolysiloxane 4.0 (aqueous phase) ion-exchanged water 50.0 1,3-butylene glycol 4.5 oligosulfation Hyaluronic acid 1.5 Sorbitan sesquioleate 3.0 Preservatives Appropriate amount Flavors Appropriate amount (Preparation method) After heating and stirring the aqueous phase, add a well-mixed and pulverized powder portion and treat with a homomixer. Further, the oil phase mixed by heating is added, and the mixture is subjected to a homomixer treatment. Then, a fragrance is added with stirring, and the mixture is cooled to room temperature.

[0043]

As described above, the external preparation for skin of the present invention not only suppresses keratinization of the skin but also suppresses the infiltration of inflammatory lymphocytes into skin tissues by the selectin mediated by the adhesive factor. It has an effect on dullness, atopic skin, contact and irritable dermatitis, sensitive skin, and dry skin, and can maintain a youthful and healthy skin condition.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI // A61K 7/035 A61K 7/035 (72) Inventor Chiaki Tanaka 1050 Nippacho, Kohoku-ku, Yokohama-shi, Kanagawa Shiseido Daiichi Co., Ltd. Inside the research center

Claims (3)

[Claims] 1. An external preparation for skin containing oligosulfated hyaluronic acid as an active ingredient. 2. The oligosulfated hyaluronic acid is of the formula (I) (Wherein, R ¹ , R ² , R ³ and R ⁴ each independently represent a hydrogen atom or an SO ₃ H group, and n is any of those having a viscosity average molecular weight of 1,000 to 50,000. And the SO ₃ H groups account for 10-90% of the total number of R ¹ , R ² , R ³ and R ⁴ )
2. The external preparation for skin according to claim 1, comprising a repeating unit represented by the formula: 3. The external preparation for skin according to claim 1 or 2, for suppressing keratinization of the skin.