JP3172599B2 - Cell activator - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cell activator which has an excellent cell activating action, is effective for improving trauma, cracks, irritations and the like and for promoting wound healing.

[0002]

2. Description of the Related Art
As the active ingredient of skin external preparations used for cell activation and wound treatment, i.e., treatment of cut wounds, treatment of wounds after shaving, cracks, irritations, sores, hemorrhoids, burns, etc., generally, allantoin and its derivatives , Sicon extract, aloe extract, ginseng extract, placenta extract and the like are known.

[0003] However, these medicinal components cannot provide a sufficient effect, and therefore, a cell activator having a remarkable cell activating effect has been desired.

[0004]

Under these circumstances, the present inventors have conducted intensive studies and as a result, have found that a remarkable cell activating effect is exhibited by combining an asparagus extract with a specific compound. The present invention has been completed.

That is, the present invention provides the following components (A) and (B): (A) an asparagus extract; (B) a yeast extract; a lactic acid bacterium extract; a bifidobacterium extract; One or two selected from beef spleen extract, reindeer muscle enzyme digest, cockscomb enzyme digest, royal jelly, pearl protein extract, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, succinic acid and derivatives thereof It is intended to provide a cell activator comprising at least one species.

The asparagus extract used in the cell activator of the present invention is asparagus (Asparagus of asparagus).
ficinalis L. ), Obtained by extracting from the stem, rhizome, leaves, flowers, etc., and its preparation method is not particularly limited. For example, the extraction is carried out at room temperature to heating using various appropriate solvents. Examples of the extraction solvent include water; lower monohydric alcohols such as methyl alcohol and ethyl alcohol; liquid polyhydric alcohols such as glycerin, propylene glycol and 1,3-butylene glycol; lower alkyl esters such as ethyl acetate; benzene, hexane and the like. Or one or more of ethers such as diethyl ether. In particular, one or a mixture of two or more of water, ethyl alcohol, glycerin, and 1,3-butylene glycol is preferable. As the extraction conditions, a solvent is used in a volume ratio of 1 to 1000 times, particularly 5 to 100 times the volume ratio of asparagus, and 4 ° C. or more, particularly 15 to 100 times.
It is preferably carried out at a temperature of 30 ° C. for 1 hour or more, particularly 1 to 3 days.

[0007] The asparagus extract obtained under the above conditions may be used as it is as an extracted solution, but if necessary, those which have been subjected to treatments such as concentration and filtration may be used as appropriate.

When the cell activator of the present invention is incorporated into an external preparation for skin, the amount of the asparagus extract is 0.0001 to 10.0% by weight (hereinafter referred to as dry solid content).
(Indicated simply by “%”), particularly preferably 0.01 to 5.0.
% Is preferred. If the content is less than 0.0001%, the effect is not sufficiently exhibited, and if it exceeds 10.0%, no further increase in the effect is observed.

In the component (B) of the present invention, the yeast extract is obtained by treating yeast itself such as baker's yeast or brewer's yeast, or a dried and powdered yeast by physical or biochemical means. Aqueous extract containing amino acids, peptides, organic acids, nucleic acids and the like. Specific examples include yeast extract A-33 (manufactured by Asahi Beer).

[0010] The lactic acid bacteria extract is produced, for example, by inoculating a lactic acid bacterium into a culture medium mainly composed of animal milk such as cow milk and performing lactic acid fermentation, and fractionating whey from the obtained culture. Lactobacillus, Acidophilus, Streptococcus thermophilus and the like can be used as lactic acid bacteria. Specific examples include Spiron-L (manufactured by Sanshou Pharmaceutical Co., Ltd.) and the like.

The bifidobacterium extract is obtained, for example, by washing a surface culture of bifidobacterium (Lactobacillus bifidum) with physiological saline and inactivating it by sonication. As a specific example, culture B. B. (Manufactured by Kotobuki Chemical Co., Ltd.).

The bovine blood deproteinized protein is obtained by subjecting the blood of an adult cow or a calf to an appropriate treatment and removing the protein. The production method is not particularly limited, but, for example, blood as a raw material may be collected from a calf that has activated the intraretinal system, or may be fresh blood of a slaughtered cow. The treatment of bovine blood is performed by heat treatment, enzymatic decomposition, electrolysis, etc., in addition to freezing treatment. The protein can be removed by an ultrafiltration method or a precipitation method, and is not particularly limited as long as it is a commonly used method. Examples of commercially available bovine blood protein deproteinization include Stimusel (Pentafarm), Sorcoseryl (Torishi Pharmaceutical) and the like, which can be suitably used. In the present invention, the bovine blood deproteinizing agent can be used as is or as a solid extract.

The bovine spleen extract is obtained, for example, by treating a water extract obtained by homogenizing fresh spleen collected from a cow. Specific examples include Revitalin P (Nikko Chemicals).

[0014] Reindeer muscle enzyme hydrolyzate is, for example, an aqueous extract obtained by physically and biochemically treating fresh skeletal muscle collected from reindeer, and contains various low molecular weight peptides and the like. is there. Specific examples include lenapeptone (manufactured by Yukon Japan).

The enzymatic decomposition product of the cockscomb is an aqueous extract obtained by enzymatically decomposing fresh chicken scallop and contains various amino acids, low molecular weight polypeptides, polysaccharides and the like. A specific example is Fibra N (manufactured by Sansho Pharmaceutical Co., Ltd.). Royal jelly is a product obtained by purifying secretions from the pharyngeal gland of worker bees, and contains various amino acids, vitamins, minerals, and the like.

The pearl protein extract is, for example, an aqueous extract of a proteolysate obtained by finely processing the shell or pearl of pearl oysters and chemically treating the fine powder, and containing various amino acids and polypeptides. Is what you do. As a specific example,
Pearl Calc extract (manufactured by Maruzen Pharmaceutical Co., Ltd.) and the like.

Adenosine triphosphate, adenosine diphosphate, adenosine monophosphate (hereinafter ATP, AD
P, AMP) and succinic acid are substances involved in the energy transfer system and the production system in vivo. In the present invention, in addition to these ATP, ADP, AMP and succinic acid, derivatives such as salts and esters thereof can be used.
For example, the monosodium salt of ATP, ADP or AMP,
Disodium salt, trisodium salt, monopotassium salt, dipotassium salt or tripotassium salt, calcium salt of succinic acid,
Disodium salts, monomethyl esters and dimethyl esters are exemplified.

These components (B) can be used alone or in combination of two or more. When the cell activator of the present invention is incorporated into an external preparation for skin, the amount of component (B) is as follows: From the viewpoint of the cell activating action and the temporal stability of the external preparation for skin, 0.001 to 20% of the total composition, particularly 0.01 to 20%.
10% is preferred.

Further, in addition to the cell activator of the present invention, aqueous components, powders, surfactants, oils, moisturizers, alcohols, and the like used in ordinary skin external preparations are included in the skin external preparations.
pH adjusters, preservatives, pigments, antioxidants, UV absorbers,
Thickeners, fragrances, cosmetic ingredients and the like can be appropriately compounded as needed. In addition, known drugs having a cell activating effect, such as allantoin and its derivatives, sicon extract, aloe extract and the like may be added.

The above-mentioned external preparation for skin can be produced according to a conventional method by blending the cell activator of the present invention. And
Emulsions, creams, lotions, serums, cleansers, packs, cleansers, foundations, etc., and can be applied as pharmaceutical, quasi-drug or cosmetic skin external preparations such as dispersions, granules, ointments, etc. it can.

[0021]

EXAMPLES The present invention will be described in more detail with reference to Test Examples and Examples, but the present invention is not limited thereto.

Production Example 1 (Asparagus Extract) Raw white asparagus root portion was 2365 as a raw material
0 g (as fresh weight) were weighed, then adjusted to an alcohol concentration of 60% and ethanol was added.
These were pulverized using a food mixer (National MX-M3) to obtain a mixture. Next, the mixture composed of these raw materials and the solvent was placed in a polybucket, and allowed to stand at room temperature for 24 hours to perform extraction. Then, the extract was collected by suction filtration in a suction filtration bottle. To the extraction residue, 25 l of 60% ethanol was added again, and the above operation was repeated. Then, about 66 l of the extract was concentrated using a rotary evaporator, and the solvent was distilled off to obtain 1000 g of a brown concentrated extract (dry solid content 4
%) Obtained (first step).

Then, until all the concentrated extracts are dissolved, n
-Add a mixed solvent of butanol and water (1: 1), shake well, and centrifuge at 10,000 rpm for 30 minutes with a centrifuge to extract and separate saponin components and the like into n-butanol layer. (Second step). Next, the n-butanol layer was concentrated by a rotary evaporator and the solvent was distilled off to obtain about 110 g of an extract (third step).

Next, water and benzene were added to the extract in an equivalent amount, and the extract was suspended to obtain a milky white solution. The solution is centrifuged at 10,000 rpm for 30 minutes using a centrifuge, and the separated benzene layer is tilted by a centrifuge tube to discard the upper layer of benzene, or the pipette tube is used to suck and remove only the upper layer. The lipid component in the extract was removed, and the same operation was performed by adding the same amount of fresh benzene to the remaining aqueous layer. In the degreasing step, only benzene alone may be added, but it has been confirmed that efficient degreasing can be performed by using another solvent such as chloroform or ether (fourth step).

Then, water was concentrated from the obtained aqueous layer portion by a rotary evaporator and dried, and then n-
About 1 liter of a mixed solvent of butanol and water (1: 1) was added to dissolve the extract, and the mixture was allowed to stand in a separating funnel for 24 hours to extract the saponin component into the butanol layer. Next, the butanol layer was concentrated and dried by a rotary evaporator to obtain about 18 g of an extract (brown extract) (fifth step).

Next, 200 ml of methanol was added to the brown extract to dissolve it, and 75 ml of the solution was slowly added dropwise to a separately prepared ethyl ether (2 l) using an injection needle to form a white precipitate. After allowing the mixture to settle, most of the ether was removed by the decantation method, and the precipitate was collected by suction filtration. The precipitate was washed several times with fresh ether to wash away the ether in which the impurities were dissolved. The substantially white precipitate obtained by such an operation was dried in a vacuum desiccator and then pulverized in a mortar to obtain about 11 g of asparagus saponin powder (sixth step).

Test Example 1 Cell growth promotion test: Using Eagle's MEM medium containing 1% fetal calf serum, and adding the samples shown in Table 1, the effect of each sample on the proliferation of mouse-derived glandular blast cells was determined. evaluated. That is, 2 × 10 ⁴ cells, which had been subcultured for a certain period of time, were seeded on a 3.5 cm-diameter plastic petri dish containing a sample-added medium, and cultured for 4 days. Thereafter, the cells were detached from the Petri dish with trypsin to prepare a cell suspension, and the number of grown cells was counted to calculate the multiplication factor. Table 1 shows the results.

[0028]

[Table 1]

As is apparent from Table 1, when the asparagus extract and each extract were combined, a remarkable cell growth promoting effect was observed as compared with the case where each was used alone.

Example 1 Cream: A cream having the composition shown in Table 2 was prepared and evaluated for the effect of improving skin roughness. Table 2 shows the results. (Production method) A. (8) to (14) are mixed by heating and kept at 70 ° C. B. (1) to (7) are mixed by heating and maintained at 70 ° C. C. Add B to A, mix and emulsify uniformly. D. After cooling C, (15) was added and mixed uniformly to obtain a cream.

(Evaluation method) The cream of the product 1-4 of the present invention or the cream of the comparative product 1-5 was applied to one face, and the cream of the comparative product 6 was applied to the other half. Was applied once a day for 2 weeks each. Two weeks later, facial skin replicas were collected and evaluated according to the criteria shown in Table 3. The value obtained by subtracting the score of the replica of the cream application site of the comparative product 6 from the obtained score of the replica of the cream application site of the present invention products 1 to 4 and comparative products 1 to 5 was defined as the degree of skin roughness improvement. In addition, the score before the start of the test is 1 on the panel.
Or I chose 2 men.

[0032]

[Table 2]

[0033]

[Table 3]

As is clear from Table 2, the creams of the products 1 to 4 of the present invention exhibited an excellent effect of improving rough skin, and a synergistic effect of the asparagus extract and the yeast extract and the like was recognized.

[0035]

Example 2 Lotion: (Formulation) (%) (1) Polyoxyethylene hydrogenated castor oil (60EO) 1.0 (2) Ethyl alcohol 10.0 (3) Preservative 0.1 (4) Appropriate amount of flavor (5) Asparagus extract ^{* 1} 1.0 (6) Lactic acid bacteria extract 0.5 (7) Sorbitol (70% aqueous solution) 3.0 (8) Horsetail extract 0.1 (9) Pyrrolidone Sodium carboxylate 3.0 (10) Purified water

(Production method) A. (1) to (4) are heated and mixed and dissolved. B. (5) to (10) are heated and mixed and dissolved. C. A and B were mixed and made uniform to obtain a lotion.

[0037]

Example 3 Emulsion: (Formulation) (%) (1) Polyoxyethylene sorbitan monostearate (10EO) 1.0 (2) Polyoxyethylene sorbitan tetraoleate (60EO) 0.5 (3) Glycerin monostearate 1.0 (4) Stearic acid 0.5 (5) Behenyl alcohol 0.5 (6) Refined avocado oil 4.0 (7) Glyceryl tri-2-ethylhexanoate 4.0 (8) Vitamin E 0.1 (9) Preservative 0.1 (10) Asparagus extract ^{* 1} 1.0 (11) Witch hazel extract 0.1 (12) Xanthan gum (2% aqueous solution) 7.0 ( 13) 1,3-butylene glycol 5.0 (14) Bovine spleen extract 1.0 (15) Enzymatic degradation product of reindeer muscle 1.0 (16) Remaining purified water (17) Appropriate amount of flavor

(Production method) (10) to (16) are mixed by heating and kept at 70 ° C. B. (1) to (9) are mixed by heating and maintained at 70 ° C. C. Add B to A, mix and emulsify uniformly. D. After cooling C, (17) was added and mixed uniformly to obtain an emulsion.

[0039]

Example 6 Ointment: (Formulation) (%) (1) Stearic acid 18.0 (2) Cetanol 4.0 (3) Triethanolamine 2.0 (4) Glycerin 5.0 (5) Asparagus Gas extract ^{* 1} 2.0 (6) Bifidobacterium extract 1.0 (7) Photosensitizer 301 0.002 (8) Garlic extract 1.0 (9) Purified water

(Production method) A. A part of (3), (4) and (9) is heated and mixed, and 7
Keep at 5 ° C. B. (1) and (2) are mixed by heating and kept at 75 ° C. C. Add A slowly to B. D. C was dissolved in the remainder of (9) while cooling (5)-
(8) was added to obtain an ointment.

Test Example 2 Wound healing test: W at 8 weeks of age
Male istar rats were subjected to the experiment in groups of 5 rats. After shaving the back of the rat, under anesthesia, the skin was incised over 4 cm in two places on the left and right so as to be symmetrical about the midline,
One is a drug application site and the other is a control site. Immediately, suture the three incisions at three places and clean with ethanol for disinfection. Of the suturing portions, one of the products 5 to 8 of the present invention or comparative products 7 to 11 (dissolved in physiological saline) shown in Table 7 was used at the drug application site, and 0.1 ml of physiological saline was used at the control site. Each was applied twice a day for one week.

One week later, the skin on the back was peeled off, and a strip-shaped section centered on the incision was made. The tensile strength of the skin section was measured using a rheometer NRM-2002J (Fudo Kogyo Co., Ltd.)
Was used for the measurement. From the obtained measured values, the wound healing rate was calculated by the following equation. Table 7 shows the results.

[0043]

(Equation 1)

[0044]

[Table 7]

As is clear from Table 7, when the components (A) and (B) were combined, the wound healing rate was clearly higher and the synergistic wound healing was higher than when each was used alone. A promoting effect was observed.

Example 5 Cream: A cream having the composition shown in Table 8 was produced, and the effect of improving skin roughness was evaluated in the same manner as in Example 1. Table 8 shows the results. (Production method) A. (11) to (18) are mixed by heating and maintained at 70 ° C. B. (1) to (10) are mixed by heating and maintained at 70 ° C. C. Add B to A, mix and emulsify uniformly. D. After cooling C, (19) was added and mixed uniformly to obtain a cream.

[0047]

[Table 8]

As is clear from Table 8, the products of the present invention 9 to 12
This cream exhibited an excellent skin roughness improving effect, and a synergistic effect of the components (A) and (B) was recognized.

[0049]

Example 6 Lotion: (Formulation) (%) (1) Polyoxyethylene hydrogenated castor oil (60EO) 1.0 (2) Ethyl alcohol 15.0 (3) Preservative 0.1 (4) Succinic acid dimethyl ester 0.1 (5) Perfume appropriate amount (6) Asparagus extract ^{* 1} 1.0 (7) Mannit 1.0 (8) Horse chestnut extract 0.5 (9) Citric acid 0 .1 (10) sodium citrate 0.3 (11) 1,3-butylene glycol 4.0 (12) purified water

(Production method) (1) to (5) are heated and mixed and dissolved. B. (6) to (12) are heated and mixed and dissolved. C. A and B were mixed to obtain a lotion.

[0051]

Example 7 Emulsion: (Formulation) (%) (1) Polyoxyethylene sorbitan monostearate (10EO) 1.0 (2) Polyoxyethylene sorbitan tetraoleate (60EO) 0.5 (3) Glycerin monostearate 1.0 (4) Stearic acid 0.5 (5) Behenyl alcohol 0.5 (6) Squalane 8.0 (7) Grape seed oil 5.0 (8) Preservative 0 .1 (9) Asparagus extract ^{* 1} 0.5 (10) ATP disodium salt 0.05 (11) Carboxyvinyl polymer 0.1 (12) Sodium hydroxide 0.05 (13) Ethyl alcohol 5.0 (14) Remaining amount of purified water (15) Suitable amount of fragrance

(Production method) A. (10), (12) and (14) are mixed by heating.
Keep at 0 ° C. B. (1) to (9) are mixed by heating and maintained at 70 ° C. C. Add B to A, mix and emulsify uniformly. D. After cooling C, (11), (13) and (15) were added and mixed uniformly to obtain an emulsion.

[0053]

Example 8 Ointment: (Formulation) (%) (1) Stearic acid 18.0 (2) Cetanol 4.0 (3) Triethanolamine 2.0 (4) Glycerin 5.0 (5) Asparagus Gas extract ^{* 1} 1.0 (6) ADP 1.0 (7) Photosensitizer 401 0.002 (8) Lysozyme chloride 1.0 (9) Purified water

(Production method) A part of (3), (4) and (9) is heated and mixed, and 7
Keep at 5 ° C. B. (1) and (2) are mixed by heating and kept at 75 ° C. C. Add A slowly to B. D. C was dissolved in the remainder of (9) while cooling (5)-
(8) was added to obtain an ointment.

[0055]

As described in detail above, the cell activator of the present invention has an excellent cell activating effect, and is effective for improving skin roughness due to trauma, cracks, nicks, etc., and promoting wound healing.

Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 7/00 A61K 7/00 W 31/70 31/70 35/14 35/14 Z 35/28 35/28 35/34 35/34 35 / 64 35/64 35/74 35/74 C 35/78 35/78 V A61P 43/00 107 A61P 43/00 107 (72) Inventor Shin Asano 48-18 Sakaemachi, Kita-ku, Tokyo KOSE RESEARCH CO., LTD. (56) References JP-A-3-86809 (JP, A) JP-A-3-236319 (JP, A) JP-A-3-11009 (JP, A) JP-A-3-1577311 (JP, A) (58) Field surveyed (Int.Cl. ⁷ , DB name) A61K 7/48 A61K 7/00 A61K 31/70 A61K 35/14 A61K 35/28 A61K 35/34 A61K 35/64 A61K 35/74 A61K 35 / 78 A61P 43/00 107

Claims (1)

(57) [Claims] 1. The following components (A) and (B), (A) asparagus extract, (B) yeast extract, lactic acid extract, bifidobacterium extract, bovine blood deproteinizing protein, and bovine spleen extract , Reindeer muscle enzyme digest, cockscomb enzyme digest, royal jelly, pearl protein extract and one or more selected from adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, succinic acid and derivatives thereof Cell activator.