JP2000309521A - Skin lotion - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a skin lotion having anti-aging effect and crease suppressing effect, etc., of the skin by including a decomposition product of collagen or gelatin with collagenase. SOLUTION: This skin lotion contains a decomposition product of collagen or gelatin with collagenase [e.g. Collagen.tripeptide M series (commercial name)] containing a peptide having an amino acid sequence of (Gly-X-Y)n (Gly is glycine; X and Y are each an amino acid other than glycine; (n) is a positive integer) and having an average molecular weight of 280-20,000. The peptide is preferably a tripeptide and the content of the peptide is preferably 30-100 wt.% based on the collagenase decomposition product. The amount of the collagenase decomposition product is preferably 0.00001-20 wt.%. The skin lotion can be used as a face lotion, a milky lotion, etc., and may be incorporated with tea extract, L-lysine, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD [0001] The present invention relates to an external preparation for the skin. More specifically, the present invention provides:
The present invention relates to an external preparation for skin which has an anti-aging effect and a wrinkle suppressing effect on skin, a cell growth promoting effect, a collagen production promoting effect, and a collagen glycation inhibiting effect.

[0002]

2. Description of the Related Art Human skin is mainly composed of an epidermis layer and a dermis layer from the outside. In particular, the dermis layer constituting the inside occupies 90% or more of the volume of the skin and plays a role of supporting the skin. The state of the dermis layer has a great effect on the skin surface beam and surface morphology. It is known that the dermis thins with age, and the cause of skin aging phenomena, such as wrinkles and sagging of the skin, is largely related to the fact that the dermis part becomes thinner and weakens It is considered.

[0003] For the purpose of preventing such aging of the skin, several attempts have conventionally been made to improve the condition of the skin by using peptides for the skin. For example, it is known that cosmetics containing oligopeptides among peptides have good adhesion to the skin and prevent skin aging such as suppressing wrinkles (Japanese Patent Application Laid-Open No. 57-57). No. 2213), especially Arg-Gl
It has been reported that two types of peptides, y-Asp-Ser and Arg-Gly-Asp, act on the stratum corneum of the skin to improve dry skin and prevent aging of the skin (Japanese Unexamined Patent Publication (Kokai) No. 2). -178207).

[0004]

As described above, the decrease in the dermis associated with aging is largely due to the decrease in the number and activity of fibroblasts supplying matrix components of the dermis. As fibroblasts decrease, matrix components, especially collagen, decrease. In addition, the remaining collagen undergoes qualitative changes such as saccharification with aging, and its function deteriorates. Therefore, exerting an improving effect on the dermis portion is a very important condition for an external preparation for skin intended to improve skin aging by suppressing wrinkles and giving skin scalp.

[0005] However, the action of the above-mentioned oligopeptide is limited to the surface of the skin, and has not been sufficiently effective to the dermis.

In an attempt to exert an effective action on the dermis, so-called cell growth factors (such as EGF and FGF) are used.
However, skin external preparations containing peptides known to have a cell proliferation action have been provided (JP-A-61-5006, etc.), but the effect of suppressing skin aging is not always satisfactory. In addition, the effect on the human body, etc. by using cell growth factors having various physiological activities,
Special consideration was needed.

[0007] Therefore, the problem to be solved by the present invention is to act on the dermis, which has solved the above-mentioned problems, to exert an excellent effect of preventing skin aging, and to improve safety. It is to provide an excellent skin external preparation.

[0008]

Means for Solving the Problems The present inventor has made intensive studies to solve this problem. As a result, they have found that a specific peptide contained mainly in collagen and gelatin degradation products of collagenase has an excellent anti-aging effect on the skin, and completed the present invention.

That is, in the present application, the present inventor has determined that collagen or gelatin decomposed by collagenase (hereinafter also referred to as “collagenase decomposed product”), particularly, its amino acid sequence is represented by (Gly-XY) n (formula Wherein Gly represents a glycine residue, X and Y represent amino acid residues excluding the glycine residue, and may be the same or different from each other, and n represents a positive integer. And a skin external preparation (hereinafter referred to as the skin external preparation of the present invention) containing, as an active ingredient, a collagen or gelatin degradation product of collagen or gelatin containing a peptide having an average molecular weight of 280 to 20,000.

[0010] The skin external preparation of the present invention specifically includes an anti-aging effect (action) on the skin [in this specification, "anti-aging effect (action)" Effect (action) "), the skin external preparation of the present invention (in particular, the skin external preparation of the present invention of this embodiment is referred to as the anti-aging skin external preparation of the present invention), wrinkle suppressing effect, Skin external preparation which has an effect of preventing the generation of wrinkles on the skin and suppressing wrinkles (the skin external preparation of the present invention of this embodiment is particularly referred to as the skin external preparation of the present invention for suppressing wrinkles), a cell growth promoting action (External skin preparation of the present invention of this embodiment is referred to as skin external preparation of the present invention in particular), skin external preparation exhibiting collagen production promoting action (the skin external preparation of this embodiment is In particular, the collagen of the present invention That production promoter external preparation for skin), collagen glycation inhibition is observed it is the skin external preparation (the present invention external preparation for skin of this embodiment, in particular, may take the aspect of a) of the present invention collagen glycation inhibiting skin external preparation.

[0011]

Embodiments of the present invention will be described below. A. About the active ingredient of the external preparation for skin of the present invention, which is the active ingredient of the external preparation for skin of the present invention, which is a starting material of a collagenase hydrolyzate, collagen is not particularly limited,
Any of type I to type XIII collagen can be used, and mixed collagen, which is a mixture thereof, can also be used. In reality, it is assumed that collagen is a mixed type of collagen obtained from various animals and fishes. However, animals (for example, cows and pigs) and fishes (for example, Flounder, salmon, sardine, tuna, etc.) and collagen extraction sites can be bones, skins, tendons, duckfishes (fishes) and the like.

[0012] Collagen can be extracted and purified from these components using generally known methods. Specifically, for example, after crushing a tissue containing collagen such as bone, skin, tendon, and sea urchin, washing with water, extraction with a dilute salt solution, extraction with an acid or alkali solution, pepsin,
The collagen can be obtained by extraction with an enzyme such as trypsin or hyaluronidase and subjecting it to a known purification means such as salting out or dialysis to purify collagen. It is also possible to obtain “regenerated collagen” by a generally known method. It is also possible to use commercially available collagen as a starting material for a collagenase degradation product.

[0013] Gelatin is a water-soluble protein obtained by heat-extracting the above-mentioned collagen with water. In the present invention, gelatin produced by a generally known method can be used as a starting material for the above-mentioned collagenase degradation product, and a commercially available product can also be used.

The collagenase hydrolyzate can be produced by allowing collagenase to act on collagen or gelatin obtained as described above (this production step,
For the content of the obtained collagenase hydrolyzate itself, see JP-A-9-176196).

The collagenase used here is not particularly limited, but may be Clostridium histolyticum Streptomyce.
s parvulus or other bacteria, actinomycetes or fungi, etc., and an amino acid sequence specific to collagen [(Gly-XY) n (wherein Gly represents a glycine residue, and X and Y exclude glycine residues) Represents an amino acid residue, which may be the same or different, and n represents a positive integer):
This amino acid sequence is also referred to as “unique amino acid sequence”))], by using a collagenase that specifically cleaves the amino-terminal side of a glycine residue to obtain a collagenase degradation product rich in peptides having this unique amino acid sequence. It is possible and preferred.

The collagenase used herein is not limited to collagenase obtained as a natural product, for example,
It may be a modified collagenase having the above specificity, which is obtained by being modified by a protein engineering technique.

The types of amino acid residues other than glycine residues, which can be taken by X and Y, are not particularly limited. Usually, amino acid residues of naturally occurring amino acids (excluding glycine), specifically, Typically, alanine residue, valine residue,
Leucine residue, isoleucine residue, proline residue, phenylalanine residue, tryptophan residue, methionine residue, serine residue, threonine residue, cysteine residue, glutamine residue, asparagine residue, tyrosine residue, lysine residue Group, an arginine residue, a histidine residue, an aspartic acid residue, and a glutamic acid residue (in the present specification, unless otherwise specified, the amino acid residue is an L-amino acid residue). Residue).

The collagenase hydrolyzate which can be used in the present invention can be produced by a generally known method, for example,
According to the method described in Japanese Patent No. 82299, collagenase free or immobilized on an immobilized carrier such as chitovar is mixed with a batch method, a column method or a combination of these methods, preferably at a reaction temperature of 40 to 45. It can be manufactured by setting the temperature to ° C.

A peptide having a unique amino acid sequence which can be produced in this manner, most preferably the above positive integer n
The collagenase hydrolyzate enriched in the tripeptide having a value of 1 (a positive integer n is preferably as small as possible in order to exert the intended effect of the present invention) is an antigen specific to gelatin or collagen. Properties are significantly suppressed,
By blending this in an external preparation for skin, the safety of the external preparation for skin against allergic reactions and the like can be further improved.

The average molecular weight of a collagenase digest containing a peptide having a unique amino acid sequence is 280 to 2000.
0 means that the anti-aging effect and the cell growth promoting effect on the skin can be expressed in the external preparation for skin of the present invention, and the antigenicity derived from collagen and the like is eliminated, and the safety on the skin is enhanced to a higher degree. It is preferable in that it can be maintained. The minimum value 280 of the average molecular weight is a molecular weight assuming the above-mentioned tripeptide in which the positive integer n is 1, and as described above, it is preferable to take a value as small as possible near 280.

Further, the content of the tripeptide having a unique amino acid sequence (where the positive integer n is 1) in the collagenase digest is 30 to 1 relative to the collagenase digest.
00% by weight, particularly preferably 7% by weight.
0-100% by weight, very preferably 85-100% by weight, most preferably 95-100% by weight.

The above-described fractionation of a collagenase digest having a specific average molecular weight or a tripeptide having a specific amino acid sequence (where the positive integer n is 1) has a specific range. Can be carried out by using a generally known fractionation method, for example, a gel filtration method or a reverse phase chromatography method, alone or in some cases in combination.

The collagenase hydrolyzate can be produced according to the above-mentioned method and can be blended with the external preparation for skin of the present invention. However, a commercially available product such as collagen tripeptide M series (Miyagi Chemical Industry Co., Ltd.) (Manufactured by K.K.) can also be used.

The amount of the collagenase hydrolyzate in the external preparation for skin of the present invention can be appropriately selected according to the specific dosage form, application, etc. of the external preparation for skin of the present invention, and is not particularly limited. Absent. Generally, it is preferably 0.00001 to 20.0% by weight, particularly preferably 0.0001 to 10.0% by weight, based on the whole agent.
Most preferably, it is 0.001 to 1.0% by weight.

This amount is 0.000 to the whole agent.
When the amount is less than 01% by weight, it is difficult to sufficiently exert an inhibitory effect on skin aging and a cell growth effect, which is not preferable. In addition to the above, there is a strong tendency that the enhancement of the effect cannot be expected in proportion to the increase in the amount, which is not only unfavorable, but also causes a problem in the preparation.

By using the collagenase hydrolyzate as an active ingredient in this manner, an external preparation for skin which exhibits an anti-aging effect based on an excellent inhibitory action on wrinkles and sagging of the skin, in particular, wrinkles on the skin An external preparation for skin that exerts an excellent effect in prevention and suppression of wrinkles, an external preparation for skin that promotes the proliferation of skin fibroblasts, suppresses the decrease in dermis, and exerts an excellent effect on wrinkles and sagging of the skin, dermis Promotes the production of collagen in the dermis, whereby the external preparation for the skin and the collagen present in the dermis exhibiting an excellent effect on wrinkles and sagging of the skin are altered by saccharification, and the action of the original collagen in the extracellular matrix is reduced. An external preparation for skin that exerts an excellent effect on skin wrinkles and sagging by suppressing
The anti-aging skin external preparation of the present invention, the wrinkle suppressing skin external preparation of the present invention, the cell proliferation promoting skin external preparation of the present invention, the collagen production promoting skin external preparation of the present invention and the collagen saccharification inhibitor of the present invention are provided. Is done.

Further, by selecting a collagenase degraded product having a high content of a tripeptide having a unique amino acid sequence (where the positive integer n is 1), the antigenicity of gelatin and collagen was remarkably suppressed. A skin external preparation that is particularly excellent not only in the above-mentioned efficacy but also in safety is provided.

B. Regarding Specific Embodiments of External Skin Preparation of the Present Invention The external skin preparation of the present invention can take any form as an external skin preparation (cosmetics, pharmaceuticals, quasi-drugs). Water, emulsion, cream, ointment, pack, aerosol, water-oil two-layer system. It can be applied to water-oil-powder three-layer preparations, serums, gels, foundations, lipsticks, shampoos, rinses and the like. In addition, the skin external preparation of the present invention, a known component effective for skin aging, a known component effective for suppression of wrinkles, a known whitening agent, etc., depending on the specific use and purpose Can be blended. Specifically, tea extract, L-lysine, L-arginine, caffeine, tannin, verapamil, tranexamic acid, tranexamic acid derivative, licorice extract, glabridine, hot water extract of karin fruit, various crude drugs, acetic acid Whitening agents such as tocopherol, glycyrrhizic acid, glycyrrhizic acid derivatives, vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid; glucose, fructose, mannose,
Sugars such as sucrose and trehalose; vitamin A derivatives such as retinoic acid, retinol, retinol acetic acid, and retinol palmitic acid, etc., together with the above-mentioned active ingredients according to the specific embodiment of the skin external preparation of the present invention. Is possible.

Depending on the specific form, various base components and the like, specifically, liquid fats and oils, solid fats and oils, waxes, ester oils, hydrocarbon oils, silicone resins, silicones,
Anionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, lower alcohols, sterols, water-soluble polymers, sequestering agents (edetate Disodium, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, etc.), neutralizing agent,
A pH adjuster, an antibacterial agent, a fragrance, a pigment, and the like can be added to the external preparation for skin of the present invention within a range that does not impair the intended effect of the present invention. The specific dosage form of the skin external preparation of the present invention will be described in Examples described later.

[0030]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to Examples, but the technical scope of the present invention is not limited thereto. In addition, the blending amount in this example is represented by% by weight based on the blending target of the blending component, unless otherwise specified.

[Test Example] Preparation of sample 50 g of high purity gelatin was added to 1000 ml of 20 mM Tris-HC.
After dissolving in l buffer (pH 7.4) /0.1 M NaCl while heating, the mixture was cooled to 50 ° C. As an immobilized enzyme for enzymatic degradation, 100 mg of collagenase enzyme (manufactured by Washington, high purity product) was bound to 50 g of chitopearl (manufactured by Fuji Boseki) using a two-crosslinking reagent. This immobilized enzyme was packed in a double column bioreactor,
Equilibration was performed with mM Tris-HCl buffer (pH 7.4) /0.1 M NaCl. High purity gelatin prepared through the above process,
The column was applied to a column of a double collagenase enzyme-immobilized column prepared in the above step, and subjected to enzymatic degradation by a column method.

[0032] The enzyme-terminated liquid that came out of the final column was fractionated and filtered with a 0.45 µm filter. The filtrate is fractionated by gel filtration or reverse phase chromatography, and has an average molecular weight of 20,000 or more, Gly-XY (Gly is a glycine residue, X and Y are amino acid residues). Wherein X and Y may be the same or different; the same applies hereinafter) having a content of 30 to 85% by weight (average molecular weight of about 3000), and having a Gly-XY content of 85 to 10
A composition having 0% by weight (average molecular weight: about 300) was prepared. After the fractionation, each fraction was subjected to a freeze-drying treatment.

The fractions obtained here (including fractions derived from collagen described later) were used in the following Test Examples and Examples.

When an anaphylaxis test was carried out on these fractions by a known method, all fractions had extremely low antigenicity, and these fractions showed safety in skin. It was found to be very excellent (results including a collagen-derived fraction described below).

2. Cell proliferation promotion test After culturing human skin fibroblasts in DMEM medium containing 10% FBS for 24 hours, 0.5% FBS containing each sample was used.
The medium was replaced with a DMEM medium containing the cells, and the cells were further cultured for 48 hours. At the end of the culture, MTT activity was measured and used as an indicator of cell proliferation. The MTT activity when cultured only in the DMEM medium containing 0.5% FBS without adding the sample was expressed as 100%. The samples were fractions prepared in the above example with a Gly-XY content of 30-85% by weight, 85-100% by weight.
Two types of fractions were examined. As a control sample, the average molecular weight in the above example was 20,000.
A similar test was performed using the above and EGF (epidermal growth factor). Table 1 shows the results of the cell growth promotion test.

[0036]

[Table 1] Table 1 ──────────────────────────────────── Sample MTT activity (% vs. Control) ───────────────────────── concentration 0.001% concentration 0.01% concentration 0.1% ────────ゼ ラ チ ン Gelatin-degraded peptide 95% 110% 110% (average molecular weight 20,000 or more) ───────分 Gly-XY content 100% 110% 130% 30 ~ 85% by weight 分─────────────────────────────────── Gly-XY content 110% 120% 150% 85-100 weight % Fraction 分───────── EGF 95% 95% 120% ────────────────────────────────── ── Note) MTT activity is a measured value of living cells using a reaction in which dyes are reduced and colored by mitochondrial respiration in cells. The higher the value, the more cell proliferation activity is observed.

From Table 1, it was found that the fraction having a Gly-XY content of 30% by weight or more had a higher cell growth promoting effect than the control. It was also found that when the Gly-XY content was 85 to 100% by weight, the cell growth promoting activity was further improved.

In place of the gelatin in the above-mentioned example, commercially available collagen was enzymatically decomposed with the collagenase enzyme used in the same example, and this was treated in the same manner as described above to give a Gly-XY content of 30%. A fraction of 85% by weight (average molecular weight: about 3000) and a fraction of 85 to 100% by weight (average molecular weight: about 300) were prepared and subjected to the same cell growth promotion test as described above. As a result, both of these collagen-derived fractions had superior cell growth promoting activity as compared with the control. Further, when the Gly-XY content was 85 to 100% by weight, the cell growth promoting activity was further improved, which was the same as the result of the above-mentioned fraction derived from gelatin.

According to the results, the collagenase hydrolyzate, which is an active ingredient of the external preparation for skin of the present invention, has an excellent cell growth promoting action, thereby suppressing the decrease of the dermis and preventing skin wrinkles and sagging. It has become clear that excellent effects can be exhibited.

3. Collagen production promotion test Human skin fibroblasts were cultured in a DMEM medium containing 10% FBS for 48 hours, and then 0.5% FBS containing each sample was cultured.
The medium was replaced with a DMEM medium containing the cells, and the cells were further cultured for 48 hours. After the completion of the culture, the culture supernatant was collected to measure the type I collagen biosynthesis ability, and the cells were collected to measure the cell amount. The samples were fractions prepared in the above example with a Gly-XY content of 30-85% by weight, 85-100% by weight.
Two types of fractions were examined. As a control sample, a similar test was performed using vitamin C, which is known to have an excellent collagen production promoting action. Cell I
The ability of type I collagen biosynthesis is determined by the C-terminal peptide of type I procollagen secreted into the culture supernatant (Procollagen type I).
C-peptide: Abbreviated as PIP. ) Was evaluated by measuring the amount. Specifically, “Procollagen type IC-peptide
(PIP) measurement kit (Takara Shuzo Co., Ltd .: MK001) ". The amount of cells was estimated based on the amount of DNA in the cells. The amount of DNA was determined by the method of Cesar Lsbarca et al. (Analyt
ical Biochemisty, 102, 344-352, (1980))
The measurement was performed using Hoechst 33258 reagent. Table 2 shows the results of this collagen production test.

[0041]

[Table 2] Table 2 P Sample PIP amount (% Control) ───────────────────────── Concentration 0.0001% Concentration 0.001% Concentration 0.01% ──────── ──────────────────────────── Gelatin-degraded peptide 95% 98% 110% (average molecular weight 20,000 or more) ───────分 Gly-XY content 118% 126% 165% 30 ~ 85% by weight 分─────────────────────────────────── Gly-XY content is 120% 160% 185% 85-100 weight % Fraction 分──────── Vitamin C (concentration 0.2mM) 178% ───────────────────────────────── ───

Table 2 shows that fractions having a Gly-XY content of 30% by weight or more have an excellent collagen production promoting effect. It was also found that when the Gly-XY content was 85 to 100% by weight, the collagen production promoting activity was further improved.

From these results, the collagenase hydrolyzate, which is an active ingredient of the external preparation for skin of the present invention, has an excellent collagen production promoting action, and thus can exert an excellent effect on wrinkles and sagging of the skin. It became clear.

4. Collagen Saccharification Inhibition Test The inhibitory effect of the saccharification reaction on the formation of crosslinks was evaluated using the collagen gel contraction activity of human dermal fibroblasts as an index. Collagen solution (Collagen is product I of Koken Co., Ltd.
-AC was used. ) Is prepared on ice, and 1
The collagen was gelled at 37 ° C. in a 2-well plate (Corning: 25815). A test substance (1,3-butylene glycol was used as a control. L-lysine and L-arginine, which have excellent collagen saccharification inhibitory activity, were also examined. The concentration was% by weight.) After adding a glucose-6-phosphate solution (final concentration; 100 mM), the mixture was incubated at 37 ° C. for 7 days to perform a saccharification reaction (glycation).

After removing unreacted glucose-6-phosphate by washing, 1 × 10 ⁵ cells / ml of fibroblasts were inoculated on a collagen gel, and 0.25% FBS / DMEM was added.
The medium was loaded. After 3 hours, the gel was peeled off from the petri dish wall, and the collagen was contracted. Forty-eight hours later, the medium was aspirated, the diameter of the collagen gel was measured, and the area was calculated. Table 3 shows the results. The smaller the collagen area after shrinkage, the more the collagen cross-linking is normally maintained, which means that the collagen is not denatured by the saccharification reaction.

[0046]

[Table 3] Table 3 ──────────────────────────────────── Glycation Test substance After contraction Collagen area (cm ² ) な し No Control 247.8 Yes Control 379.9 Yes Gly-XY content of 30 to 85% by weight (0.05%) 290.2 Yes Gly-XY content of 85 to 100% by weight (0.05%) 268.5 Yes L-Lysine (0.5%) 243.0 Yes L -Arginine (0.5%) 241.2 ────────────────────────────────────

Table 3 shows that fractions having a Gly-XY content of 30% by weight or more show excellent collagen glycation-inhibiting action. Further, it was revealed that when the Gly-XY content is 85 to 100% by weight, the action of inhibiting saccharification of collagen is more excellent.

As a result, the collagenase hydrolyzate, which is an active ingredient of the external preparation for skin of the present invention, has an excellent inhibitory effect on collagen saccharification, and thus can exert an excellent effect on wrinkles and sagging of the skin. It became clear.
As described above, collagenase, which constitutes a major fibrous structure in the dermis layer, is quantitatively maintained (collagen production promoting activity) and qualitatively (inhibition of denaturation due to collagen saccharification) in collagenase degradation products. Action is recognized, by using it as an active ingredient of a skin external preparation,
It has been found to be effective in preventing and improving the aging phenomenon (typically wrinkles and sagging) related to the skin structure.

5. Skin aging improvement effect test Examples 1 to Comparative Example 1 described below (prepared by replacing a fraction having a Gly-XY content of 30 to 85% by weight in the formulation of Example 1 with ion-exchanged water). The actual use test shown below was performed about the cosmetics (cream).

Test Method A healthy person (female) aged 25 to 60 years, randomly selected
After using the test article on the facial skin for one month every day, 100 subjects were examined, and the effect of improving wrinkles and the effect of improving sagging were examined.

A) Evaluation of effect on wrinkles: The condition of the outer corners of the eyes was visually observed and evaluated based on the following evaluation criteria.

Evaluation criteria A: Very improved B: Improved C: Slightly improved D: No improvement effect was observed E: Deteriorated b) Effect on sagging The skin condition of the entire face was visually observed. It was observed and evaluated based on the following evaluation criteria.

Evaluation criteria A: very improved B: improved C: slightly improved D: no improvement effect was observed E: deteriorated The results of these practical use tests are shown in Table 4.

[0054]

[Table 4] Table 4 効果 Effect on wrinkles (people) Sagging Effect (person) ABCDEABCDE ─────────────────────────────────── ─ Example 1 19 32 38 11 0 22 28 32 18 0 Comparative Example 1 6 21 45 26 2 0 23 58 19 0 ─────────────────────── ─────────────

From Table 4, it is clear that when the cream of the present invention of Example 1 was used, wrinkles at the outer corners of the eyes and sagging of the skin were higher than when the cream of Comparative Example 1 was used. It is recognized that the above has been improved. This has revealed that a skin external preparation containing the above fraction as an active ingredient is an external preparation that has an anti-aging effect, a wrinkle suppressing effect, and the like on the skin.

Hereinafter, examples of the formulation of the skin external preparation of the present invention in various dosage forms, including Example 1 described above, will be described as examples. In addition, when the above-mentioned actual use test was performed also on the examples other than the example 1, in any of the examples,
Very excellent anti-aging and wrinkle-suppressing effects of the skin were observed.

Example 1 Cream Ingredient Amount (% by Weight) Stearic Acid 5.0 Stearyl Alcohol 4.0 Isopropyl Myristate 18.0 Glycerin Monostearate 3.0 Propylene Glycol 10.0 Gly-XY Content 30-85% by weight (derived from gelatin) 0.01 KOH 0.2 Sodium bisulfite 0.01 Preservatives Appropriate amount Fragrance Appropriate amount Ion-exchanged water residue <Production method> Ion-exchanged water with propylene glycol , A fraction, and KOH were added, and after dissolution, the mixture was heated and kept at 70 ° C (aqueous phase). In this aqueous phase, other components are heated and melted,
The oil phase kept at 0 ° C. was gradually added, and after all the addition was completed, the temperature was kept at that temperature for a while to cause a reaction. afterwards,
Emulsify evenly with a homomixer and, with good stirring,
Cooling to ° C gave a cream.

Example 2 Cream Ingredients Ingredients (% by Weight) Stearic Acid 2.0 Stearyl Alcohol 7.0 Hydrogenated Lanolin 2.0 Squalane 5.0 2-Octyldodecyl Alcohol 6.0 Polyoxyethylene (25 Mol) Cetyl alcohol ether 3.0 Glycerin monostearate 2.0 Propylene glycol 5.0 Gly-XY Fraction (gelatin-derived) having a content of 85 to 100% by weight 0.05 Sodium bisulfite 0.03 Ethyl paraben 0.3 Appropriate amount of perfume Ion-exchanged water residue <Production method> Propylene glycol was added to ion-exchanged water, and heated to 70 ° C (aqueous phase). To this aqueous phase, add an oil phase that is heated and melted at 70 ° C. by heating and melting other components, pre-emulsify, uniformly emulsify with a homomixer, and then cool to 30 ° C. while stirring well to obtain a cream. I got

[Example 3] Cream blending ingredients blending amount (% by weight) Solid paraffin 5.0 Beeswax 10.0 Vaseline 15.0 Liquid paraffin 41.0 Glycerin monostearate 2.0 Polyoxyethylene (20 mol) Sorbitan monolaurate 2.0 Soap powder 0.1 Borax 0.2 Gly-XY A fraction with a content of 30 to 85% by weight (derived from collagen) 0.5 Warmoko ethanol extract 0.05 Sodium bisulfite 0.03 Ethylparaben 0.3 Perfume Appropriate amount Ion-exchanged water residue <Production method> Soap powder and borax were added to ion-exchanged water, heated and dissolved, and kept at 70 ° C (aqueous phase). An oil phase maintained at 70 ° C. by heating and melting the other components was gradually added to the aqueous phase and reacted. After the completion of the reaction, the mixture was uniformly emulsified with a homomixer. After the emulsification, the mixture was cooled to 30 ° C. with good stirring to obtain a cream.

[Example 4] Emulsion compounding amount (% by weight) Stearic acid 2.5 Cetyl alcohol 1.5 Vaseline 5.0 Liquid paraffin 10.0 Polyoxyethylene (10 mol) Monooleate 2.0 Polyethylene glycol 1500 3.0 Triethanolamine 1.0 Carboxyvinyl polymer 0.05 (trade name: Carbopol 941, BFGoodrich Chemical company) Gly-XY fraction (derived from collagen) having a content of 85 to 100% by weight 0 Sodium bisulfite 0.01 Ethylparaben 0.3 Perfume Appropriate amount Ion-exchanged water residue <Production method> Propylene glycol was added to ion-exchanged water and heated to 70 ° C (aqueous phase). To this aqueous phase, the oil phase, which was heated and melted at 70 ° C. with other components, was gradually added, uniformly emulsified with a homomixer, and after emulsification, cooled to 30 ° C. with good stirring. An emulsion was obtained.

[Example 5] Jelly blending component blending amount (% by weight) 95% ethyl alcohol 10.0 dipropylene glycol 15.0 polyoxyethylene (50 mol) oleyl alcohol ether 2.0 carboxyvinyl polymer 1.0 ( Trade name: Carbopol 940, BFGoodrich Chemical company) NaOH 0.15 L-arginine 0.1 Gly-XY Fraction having a content of 30 to 85% by weight (derived from gelatin) 0.0001 2-hydroxy-4-methoxybenzophenone Sodium sulfonate 0.05 Ethylenediaminetetraacetate 3 Trisodium 2 water 0.05 Methyl paraben 0.2 Perfume Appropriate amount Ion exchange water residue <Production method> Carboxyvinyl polymer is uniformly dissolved in ion exchange water (aqueous phase) Separately, in 95% ethyl alcohol, polyoxyethylene Down was dissolved (50 mol) oleyl alcohol ether which was added to the aqueous phase.
Next, after adding other components to this, it neutralized with NaOH and L-arginine, thickened, and obtained jelly.

Example 6 Essence Composition (Weight% ) 95% Ethyl Alcohol 10.0 Polyoxyethylene (20 mol) Octyldodecanol 1.0 Pantothenyl Ethyl 0.1 Gly Preparative fraction having -XY content of 85 to 100% by weight (derived from gelatin) 1.5 Methyl paraben 0.15 (B phase) KOH 0.1 (C phase) Glycerin 5.0 Dipropylene glycol 10.0 Sodium bisulfite 0 .03 Carboxyvinyl polymer 0.2 (trade name: Carbopol 940, BFGoodrich Chemical company) Ion-exchanged water Residue <Production method> A phase and C phase are uniformly dissolved respectively, and A phase is added to C phase. Solubilized. Then, after adding phase B,
Filling was performed to obtain a serum.

Example 7 Packing Ingredients (Weight%) (A phase) Dipropylene glycol 5.0 Polyoxyethylene (60 mol) hydrogenated castor oil 5.0 (B phase) Gly-XY content is 30 -85 wt% fraction (derived from collagen) 0.01 Olive oil 5.0 Tocopherol acetate 0.2 Ethylparaben 0.2 Fragrance 0.2 (C phase) Sodium bisulfite 0.03 Polyvinyl alcohol 13.0 (Ken Ethanol 7.0 Deionized water Residue <Production method> A phase, B phase and C phase were each uniformly dissolved, and A phase was added to B phase to be solubilized. Then, this is C
After adding to the phase, filling was performed to obtain a pack.

Example 8 Solid Foundation Compounding Component Mixing Amount (% by Weight) Talc 43.1 Kaolin 15.0 Sericite 10.0 Zinc White 7.0 Titanium Dioxide 3.8 Yellow Iron Oxide 2.9 Black Iron Oxide 0.2 Squalane 8.0 Isostearic acid 4.0 POE sorbitan monooleate 3.0 Isocetyl octoate 2.0 Gly-XY Fractionated (collagen-derived) having a content of 85 to 100% by weight 1.0 Preservative Suitable Amount Perfume Appropriate amount <Production method> The talc-black iron oxide powder components are mixed well in a blender, and the squalane-isocetyl octanoate oil components, fractions, preservatives, and fragrances are added and kneaded well. The container was filled and molded to obtain a solid foundation.

[Example 9] Emulsion type foundation (cream type) compounding components compounding amount (% by weight) (powder part) titanium dioxide 10.3 sericite 5.4 kaolin 3.0 yellow iron oxide 0.8 bengara 0 0.3 Black iron oxide 0.2 (oil phase) Decamethylcyclopentasiloxane 11.5 Liquid paraffin 4.5 Polyoxyethylene-modified dimethylpolysiloxane 4.0 (aqueous phase) Deionized water 50.0 1,3-butylene Glycol 4.5 Gly-XY Fraction with a content of 30 to 85% by weight (derived from gelatin) 1.5 Sorbitan sesquioleate 3.0 Preservatives Appropriate amount Fragrance Appropriate amount <Production method> After heating and stirring the aqueous phase component Then, a powder portion sufficiently mixed and pulverized was added thereto, and a homomixer treatment was performed. Furthermore, after adding the heat-mixed oil phase and performing a homomixer treatment, adding a flavor while stirring, and cooling to room temperature,
An emulsion type foundation was obtained.

[0066]

Industrial Applicability According to the present invention, an external preparation for skin which has an anti-aging effect on skin, a wrinkle suppressing effect, a cell growth promoting effect, a collagen production promoting effect and a collagen glycation inhibiting effect, and which is extremely excellent in safety. Is provided.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) A61P 43/00 107 A61K 37/18

Claims (8)

[Claims] 1. An external preparation for skin containing as an active ingredient a decomposition product of collagen or gelatin by collagenase. 2. The degradation product of collagen or gelatin by collagenase has the amino acid sequence of (Gly-X-
Y) n (where Gly represents a glycine residue; X, Y
Represents an amino acid residue excluding a glycine residue; X and Y may be the same or different; n represents a positive integer); and an average molecular weight of 280 to 2
The external preparation for skin according to claim 1, which contains a peptide which is 0000. 3. Gly-XY (in the formula, Gly represents a glycine residue, X and Y represent amino acid residues excluding a glycine residue) in a degradation product of collagen or gelatin by collagenase, The external preparation for skin according to claim 2, wherein the content of the tripeptide represented by X and Y may be the same or different from each other) is 30 to 100% by weight in the decomposed product. 4. The skin external preparation according to claim 1, wherein the skin external preparation is an anti-aging skin external preparation. 5. The external preparation for skin according to claim 1, wherein the external preparation for skin is a wrinkle suppressing skin external preparation. 6. The external preparation for skin according to claim 1, wherein the external preparation for skin is an external preparation for skin having an effect of promoting cell proliferation. 7. The external preparation for skin according to any one of claims 1 to 3, wherein the external preparation for skin is an external preparation for skin having an effect of promoting collagen production. 8. The external preparation for skin according to any one of claims 1 to 3, wherein the external preparation for skin is an external preparation for skin in which suppression of collagen saccharification is observed.