JP4914029B2 - Type IV and type VII collagen production promoter - Google Patents

Description

  The present invention relates to a collagen production promoter, and more particularly to a collagen production promoter that promotes the production of both type IV collagen and type VII collagen among the collagen that is one of the extracellular matrix components.

  Occurrence of wrinkles and sagging seen in aging skin is a major aging change in appearance, and is a serious problem for many middle-aged and elderly people. One of the causes of wrinkles and sagging is due to thinning of skin tissue with aging. In aging skin, the decrease in collagen fibers, which are the main matrix component of the dermis, is significant, which is the main cause of the reduction in skin thickness.

  These collagen fibers include type IV collagen and type VII collagen. Both type IV and type VII collagens are components of the epidermal basement membrane that play a role in binding the epidermis and dermis. Type IV collagen is a major component of the lamina densa structure that forms the skeleton of the epidermal basement membrane. In addition, type VII collagen is the main component of anchoring fibers that bind the basement membrane to the dermis.

  The level of expression of type IV collagen in the epidermal basement membrane has been observed to decrease with age (Vazquez F et al., Maturitas 1996, 25: 209-215). Report that dermal fibroblasts derived from humans have reduced productivity at the protein and mRNA levels compared to dermal fibroblasts derived from young people (Chen et al., J. Invest. Dermatol., 102: 205-209 , 1994). In addition, there is a report that anchoring fibers composed of type VII collagen decrease with physiological aging and photoaging in normal skin (Takuo Tsuji, Nisshinkai 105: 963-975, 1995, Tidman et al., J Invest Dermatol., 83: 448-453, 1984). Therefore, promoting the production of type IV collagen that forms the basement membrane skeleton, anchoring fiber formation that binds the epidermis and dermis, or type VII collagen, the main component of anchoring fiber, It is extremely effective in maintaining such healthy and youthful skin and preventing and improving wrinkles and sagging.

  Conventionally, as an ingredient derived from a natural product that promotes collagen production to prevent / improve aging changes of the skin, for example, isoflavone compounds and phytosterols selected from soybean zein, soybean gin, genistein, and genistin have been reported. (See Patent Document 1).

JP 2001-39849 A

  An object of the present invention is to provide a collagen production promoter that promotes the production of both type IV collagen and type VII collagen.

  Therefore, as a result of intensive research on substances having an action of promoting the production of type IV and type VII collagens, the present inventor is prominent with respect to the fact that the solvent extract of Yuzu seeds promotes the production of type IV and type VII collagen. It has been found that it is effective.

  That is, the present invention provides a type IV collagen production promoter characterized by comprising a solvent extract of yuzu seeds, and a type VII collagen production promoter characterized by comprising a solvent extract of yuzu seeds Is. Moreover, according to this invention, the collagen production promotion method characterized by promoting the production of collagen using the said collagen production promoter is provided.

  Furthermore, the present invention is a skin external preparation for promoting collagen production, characterized in that the collagen production promoter is blended.

  The present invention is also a laminin-5 production promoter characterized by comprising a solvent extract of yuzu seeds.

  Yuzu fruit extract is known to promote hyaluronic acid production (Japanese Patent Laid-Open No. 2001-158728). Yuzu seed extract has a collagen production promoting action and a laminin 5 production promoting action. It is not known to have. Further, having a hyaluronic acid production promoting effect, a collagen production promoting effect, and having a laminin 5 production promoting effect are different technical matters. It does not necessarily have a production promoting effect, particularly a type IV or type VII collagen production promoting effect and a laminin 5 production promoting effect.

  The collagen production promoter of the present invention is excellent in the effect of promoting both type IV and type VII collagen production and is safe. Therefore, the collagen production promoter and the skin external preparation for promoting collagen production of the present invention can promote the production of both type IV collagen and type VII collagen and maintain the amount of type IV and type VII collagen. It is effective in preventing and improving sagging.

  Furthermore, type IV collagen and type VII collagen constitute a structure necessary for adhesion to the epidermis, and play an important role in the adhesion between the epidermis and the dermis. It is also useful for the treatment of blisters (blisters) characterized by blister formation directly below.

  The laminin 5 production promoter of the present invention is excellent in the action of promoting the production of laminin 5, promotes the production of laminin 5 in the skin, can maintain the amount of laminin 5, and can prevent and improve wrinkles and sagging. It is valid.

The present invention is described in detail below.
The collagen production promoter of the present invention comprises a solvent extract of Yuzu seeds.
The plant extract can be obtained by a conventional method. For example, the plant extract can be obtained by immersing or heating and refluxing the plant with an extraction solvent, followed by filtration and concentration. As the extraction solvent, any solvent that is usually used for extraction can be used. For example, water, methanol, ethanol, propylene glycol, 1,3-butylene glycol, glycerin and other alcohols, hydrous alcohols, chloroform , Organic solvents such as dichloroethane, carbon tetrachloride, acetone, ethyl acetate, hexane and the like can be used alone or in combination. The extract obtained by extraction with the above solvent is adsorbed as it is, or the concentrated extract is adsorbed by an adsorption method, for example, by removing impurities using an ion exchange resin, or by a column of a porous polymer (eg Amberlite XAD-2). Thereafter, a product eluted with methanol or ethanol and concentrated can also be used. Further, a partitioning method, for example, an extract extracted with water / ethyl acetate can be used.

  For example, the collagen production promoter of the present invention can be obtained by crushing a seed of yuzu, using water-containing ethanol or water as a solvent, and immersing or heating at room temperature to extract. As the solvent, 75 to 99% aqueous ethanol is most preferable.

  Said collagen production promoter can be mix | blended with another component and can be used as the skin external preparation for collagen production promotion.

  When the collagen production promoter of the present invention comprising the above-mentioned yuzu seed extract is used in combination with the skin external preparation for promoting collagen production of the present invention, it is 0.0005 to 20.0 mass as a dry weight in the total amount of the external preparation. %, Preferably 0.001 to 10.0% by mass. If it is less than 0.0005% by mass, the effect of promoting collagen production of the present invention is not sufficiently exhibited. On the other hand, if it exceeds 20.0% by mass, it is difficult to make a preparation, which is not preferable. Moreover, even if it mixes exceeding 10.0 mass%, the improvement of the big effect is not recognized.

  In addition to the above-described collagen production promoter, the skin external preparation for promoting collagen production according to the present invention is generally used for skin external preparations such as cosmetics and pharmaceuticals, such as whitening agents, moisturizers, antioxidants, oily components, Ultraviolet absorbers, surfactants, thickeners, alcohols, powder components, color materials, aqueous components, water, various skin nutrients, and the like can be appropriately blended as necessary.

  Others, disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, sequestering agents such as gluconic acid, caffeine, tannin, verapamil, tranexamic acid and its derivatives, licorice extract, grabrizine , Hot water extract of fire thorn fruit, various herbal medicines, drugs such as tocopherol acetate, glycyrrhizic acid and its derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, glucoside ascorbate, arbutin, kojic acid, etc. Whitening agents, sugars such as glucose, fructose, mannose, sucrose, trehalose, vitamin A derivatives such as retinoic acid, retinol, retinol acetate, retinol palmitate, higi extract, beech extract, tonin extract, hypotaurine, silica Covered zinc oxide, overnight Kusou extract, aminomethyl cyclohexane carboxylic amide, xylitol, arginine, serine, trimethyl glycine, alpha-glucosyl hesperidin, apricot extract, hydroxyproline, etc. may also be suitably incorporated.

  The skin external preparation for promoting collagen production of the present invention can be widely applied to cosmetics, quasi-drugs, etc., particularly preferably cosmetics applied to the outer skin. A wide range of dosage forms such as a solubilizing system, an emulsifying system, a powder system, an oil liquid system, a gel system, an ointment system, an aerosol system, a water-oil two-layer system, and a water-oil-powder three-layer system can be adopted. That is, if it is a basic cosmetic, it can be widely applied in the above-mentioned various dosage forms in the form of face wash, lotion, milky lotion, cream, gel, essence (beauty liquid), pack, mask and the like. In addition, makeup cosmetics can be widely applied to foundations and the like, and toiletries such as body soaps and soaps. Furthermore, if it is a quasi-drug, it can be widely applied to various ointment forms. And the form which the collagen production promoter of this invention can take is not limited to these dosage forms and forms.

  Moreover, according to this invention, the laminin 5 production promoter characterized by consisting of the solvent extract of the said seed of Yuzu is provided. Details of the solvent extract of Yuzu seeds in the laminin 5 production promoter and the application method thereof, and the application method of the laminin 5 production promoter are the same as in the collagen production promoter.

  Hereinafter, the present invention will be described in more detail with reference to examples. In addition, this invention is not limited by this. A compounding quantity is the mass%. Prior to the examples, an effect test method and results will be described.

Test example 1
(1-1) Preparation of Yuzu Seed Extract 100 g of Yuzu seeds were pulverized and extracted by heating and reflux extraction using 1000 ml of a 90% aqueous ethanol solution. The obtained extract was concentrated to obtain 836 ml of a water-containing ethanol extract of citron seeds.

(1-2) Collagen production promotion effect test Human skin fibroblasts (hereinafter referred to as cells) were used to evaluate the ability of cells to biosynthesize collagen.

(Type IV and VII collagen assay methods)
(1) Culture of human fibroblasts Human fibroblasts cultured in DMEM medium containing 10% FBS are seeded in a 24-well plate, and after the cells adhere, they are added to DMEM medium containing 0.25% FBS and 250 μM ascorbic acid glucoside. Replaced and added Yuzu seed extract. One day later, the supernatant of the medium was collected and centrifuged, and the type IV and type VII collagens in the obtained supernatant were measured, and the amount of DNA was measured for the cells, which was used as an indicator of the number of cells.

(2) DNA quantification The amount of DNA was measured by a fluorescence measurement method using Hoechst H33342. It should be noted that the concentration of the aqueous solution of the seed extract of Yuzu is 1 × 10 ⁻³ mass%, 3 × 10 ⁻³ mass%, and 1 × 10 ⁻² mass%, which has no effect on cell growth. It was confirmed that there was no cytotoxicity.

(3) Measurement of type IV and type VII collagen by sandwich ELISA
Type IV and type VII collagen were measured by sandwich ELISA. The antibodies used in this example are as follows.
Type IV collagen specific antibody; monoclonal antibody JK-199 and polyclonal antibody MO-S-CLIV
Type VII collagen specific antibody; monoclonal antibody NP-185 and monoclonal antibody NP-32
When the amount of type IV and type VII collagen per sample of the sample (control) to which no seed extract of yuzu was added was 100, the amount of type IV and type VII collagen per DNA of the added sample was expressed as type IV, The rate of type VII collagen production was promoted.

(1-3) Hyaluronic acid production promoting effect test of epidermis (keratinocytes) (1) Hyaluronic acid production promoting test method The plant extract obtained in (1-1) above was added to dimethyl sulfoxide (DMSO) at a concentration of 10%. It dissolved so that it might become each plant extract containing solution (henceforth "original solution"). A solution whose concentration was adjusted by diluting this original solution was used as a test solution, and the following experiment was conducted.
Human skin-derived immortalized epidermal cells were seeded at 20,000 per well in a 24-well petri dish, and cultured for 4 days in a growth factor-containing KGB medium (Kurabo Co., Ltd.). Thereafter, the medium was replaced with 2 ml of KGB medium containing the above test solution, and further cultured for 4 days. The plant extract concentration in the medium was 1 × 10 ⁻³ mass%, 3 × 10 ⁻³ mass%, and 1 × 10 ⁻² mass%.
After the culture, the medium was collected and hyaluronic acid was measured. The hyaluronic acid was measured using a commercially available hyaluronic acid measurement kit (manufactured by Chugai Pharmaceutical Co., Ltd.). Further, the amount of DNA in the petri dish was measured and used as an index of the number of cells. The amount of DNA was measured by a fluorescence measurement method using “Hoechst 33258” (manufactured by Hoechst).
The hyaluronic acid production promoting action was evaluated by the hyaluronic acid production promoting rate. The hyaluronic acid production promotion rate (%) is a medium containing each plant extract when the amount of hyaluronic acid per DNA of human skin-derived immortalized epidermal cells (control) cultured in a medium not containing each plant extract is 100. It was defined as the amount of hyaluronic acid per DNA of human skin-derived immortalized epidermal cells cultured in the above.

(2) DNA quantification The amount of DNA was measured by a fluorescence measurement method using Hoechst H33342. It should be noted that the concentration of the aqueous solution of the seed extract of Yuzu is 1 × 10 ⁻³ mass%, 3 × 10 ⁻³ mass%, and 1 × 10 ⁻² mass%, which has no effect on cell growth. It was confirmed that there was no cytotoxicity.

  The results are shown in Table 1. In any case, a clear collagen production promoting effect and a hyaluronic acid production promoting effect on the epidermis (keratinocytes) were recognized as compared with the sample not added with the yuzu seed extract aqueous solution (control = 100). Table 1 also shows the results of measuring the collagen production promoting effect and the hyaluronic acid production promoting effect of the epidermis (keratinocytes) in the same manner as above except that the fruit of yuzu was used instead of the seed of yuzu.

Next, CGS27023 (N-Hydroxy-2R-[[(4-methoxyphenyl) sulfonyl] (3-picolyl) amino] -3-methylbutanamide Hydrochloride) (J Med. Chem. 1997, 40 2525-2532)), the collagen production promoting action of type IV and type VII collagen was examined in the same manner as described above. As a result, in 10 ⁻⁷ mass% and 10 ⁻⁶ mass% (IC 50 = about 9 × 10 ⁻⁷ mass% protein nucleic acid enzyme 2000,45 1083-1089) where collagenase inhibitory effect can be sufficiently expected, It was not recognized at all.
This shows that the collagenase inhibitory effect that prevents the degradation of collagen and the collagen production promoting effect that promotes the production of collagen are different functions.

(1-4) Laminin 5 production promoting effect test (1) Culture of epidermal keratinocytes Epidermal keratinocytes were isolated from human foreskin and cultured in epidermal cell growth medium (KGM) with low calcium concentration. The bovine pituitary extract and EGF were added to this medium. The cells were cultured in KGM until the fourth generation, then the adherent cells were suspended by trypsin-EDTA treatment, the cell aggregates were removed by filtration, and a uniform cell suspension was obtained. Cells were collected by centrifugation and resuspended in DMEM-F12 (2: 1) -0.1% BSA at 8 × 10 ⁴ / ml. 0.5 ml of this cell suspension was added to 0.5 ml of the same medium containing a double concentration of the drug. The culture was performed at 37 ° C. for 24 hours using a 24-well plate. At the end of the culture, the culture supernatant was transferred to an Eppendorf tube, centrifuged at 10,000 rpm for 5 minutes, the supernatant transferred to a new tube, and stored at −20 ° C. until the day of laminin 5 measurement. Further, in order to solubilize laminin 5 bound to the inside of the cell and on the culture plastic, Tris-HCl buffer (pH 7.4) containing various surfactants was added to each well and stored overnight at -20 ° C. The next day, it was sonicated and frozen again. The next day, after dissolving again, the mixture was centrifuged at 10,000 rpm for 5 minutes, and the supernatant was transferred to a tube and stored at −20 ° C. until the day of measurement of laminin 5.

(2) Measurement of laminin 5 by sandwich ELISA method Laminin 5 present in the culture supernatant and cell layer was measured by sandwich ELISA method. A BM165 monoclonal antibody against laminin α3 chain of laminin 5 was bound to a solid layer of a 96-well ELISA plate. In order to sandwich and measure laminin 5, 6F12, which is a monoclonal antibody against laminin β3 chain, was biotinylated (b-6F12) in advance as another type of antibody. In this method, only the heterotrimeric body (α3β3γ2) capable of exhibiting the function is measured, and the heterodimer (β3γ2) is not detected. Samples are added to each well previously filled with 3% gelatin / phosphate buffer solution containing b-6F12. The final dilution ratio in the sample hole was 1/4 for the culture solution and 1/10 for the cell layer. The antigen-antibody reaction was carried out at 37 ° C. for 2 hours. After the plate was washed, an avidin HRP (horseradish peroxidase) solution was added, and the reaction was further continued for 30 minutes to 1 hour. After washing, an ABTS solution which is a substrate for HRP was added, and the absorbance at 405 nm was measured with an ELISA plate reader. A calibration curve was prepared in the range of 0 to 40 ng / ml.
The amount of laminin 5 produced was calculated as the sum of the amount released in the medium and the amount remaining in the cell layer, and was expressed as a value relative to the sample (control) to which no plant extract was added. The results are shown in Table 2.

  From Table 2, it became clear that the extract of the seed of Yuzu promoted the production amount of laminin 5 depending on the concentration.

[Example 1] Cream (prescription)
Stearic acid 2.0% by mass
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 ester 2.0
Propylene glycol 5.0
Yuzu seeds 70% ethanol room temperature extract 5.0
Tranexamic acid 0.2
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Propylene glycol is added to ion-exchanged water and heated to 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). Preliminarily emulsify by adding an oil phase to the aqueous phase, uniformly emulsify with a homomixer, and then cool to 30 ° C. while stirring well.

[Example 2] Cream (prescription)
Solid paraffin 5.0% by mass
Beeswax 10.0
Vaseline 15.0
Liquid paraffin 41.0
Glycerin monostearate ester 2.0
Polyoxyethylene (20 mol) sorbitan
Monolaurate 2.0
Soap powder 0.1
Yuzu seeds 50% ethanol heat extract 0.05
Sodium bisulfite 0.03
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Add soap powder to ion-exchanged water and heat to maintain at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). The reaction is gradually added while stirring the oil phase in the aqueous phase. After completion of the reaction, the mixture is uniformly emulsified with a homomixer and cooled to 30 ° C. while stirring well after emulsification.

[Example 3] Emulsion (prescription)
Stearic acid 2.5% by mass
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
(Product name: Carbopol 941, BFGoodrich Chemical company)
Yuzu seeds 90% ethanol heated extract 0.01
Sodium bisulfite 0.01
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Dissolve the carboxyvinyl polymer in a small amount of ion-exchanged water (A phase). Polyethylene glycol 1500 and triethanolamine are added to the remaining ion-exchanged water, dissolved by heating and maintained at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). Add the oil phase to the water phase, preliminarily emulsify, add the A phase, uniformly emulsify with a homomixer, and cool to 30 ° C. while stirring well after emulsification.

[Example 4] Emulsion (prescription)
Microcrystalline wax 1.0% by mass
Beeswax 2.0
Lanolin 20.0
Liquid paraffin 10.0
Squalane 5.0
Sorbitan sesquioleate ester 4.0
Polyoxyethylene (20 mol) sorbitan
Monooleate 1.0
Propylene glycol 7.0
Yuzu seed 30% ethanol room temperature extract 10.0
Tranexamic acid 1.0
Sodium bisulfite 0.01
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Propylene glycol is added to ion-exchanged water and heated to 70 ° C. (aqueous phase). The other ingredients 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 uniformly emulsified with a homomixer. After emulsification, cool to 30 ° C while stirring well.

Example 5 Jelly (Prescription)
95% ethyl alcohol 10.0% by mass
Dipropylene glycol 15.0
Polyoxyethylene (50 mol)
Oleyl alcohol ether 2.0
Carboxyvinyl polymer 0.05
(Product name: Carbopol 940, BFGoodrich Chemical company)
Caustic soda 0.15
L-Arginine 0.1
Yuzu seed hot water extract 0.001
Sodium bisulfite 0.01
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Carbopol 940 is uniformly dissolved in ion-exchanged water, while polyoxyethylene (50 mol) oleyl alcohol ether and retinol are dissolved in 95% ethanol and added to the aqueous phase. Next, after adding other ingredients, the mixture is neutralized and thickened with caustic soda and L-arginine.

[Example 6] Jelly (prescription)
(Phase A)
Ethyl alcohol (95%) 10.0% by mass
Polyoxyethylene (20 mol) octyldodecanol 1.0
Pantotenyl ethyl ether 0.1
Yuzu seeds 99% ethanol room temperature extract 0.001
Methylparaben 0.15
(Phase B)
Potassium hydroxide 0.1
(Phase C)
Glycerin 5.0
Dipropylene glycol 10.0
Sodium bisulfite 0.03
Carboxyvinyl polymer 0.2
(Product name: Carbopol 940, BFGoodrich Chemical company)
Purified water residue (production method)
A phase and C phase are uniformly dissolved, and A phase is added to C phase to solubilize. Next, filling is performed after adding phase B.

[Example 7] Pack (prescription)
(Phase A)
Dipropylene glycol 5.0% by mass
Polyoxyethylene (60 mol) hydrogenated castor oil 5.0
(Phase B)
Yuzu seeds 75% ethanol heat extract 0.01
Olive oil 5.0
Tocopherol acetate 0.2
Ethylparaben 0.2
Fragrance 0.2
(Phase C)
Sodium bisulfite 0.03
Polyvinyl alcohol (saponification degree 90, polymerization degree 2,000) 13.0
Ethanol 7.0
Purified water residue (production method)
A phase, B phase, and C phase are uniformly dissolved, and B phase is added to A phase to solubilize. Next, this is added to phase C and then filled.

[Example 8] Sun protection cosmetic (prescription)
Stearic acid 1.5% by mass
Cetyl alcohol 3.0
Beeswax 2.0
Polyoxyethylene (10 mol addition)
Monooleate 1.0
Glycerin monostearate ester 1.0
Dimethylpolysiloxane 10.0
Decamethylcyclopentasiloxane 20.0
2-Hydroxy-4-methoxybenzophenone 3.0
Octyl p-methoxycinnamate 2.0
Yuzu seed 90% ethanol room temperature extract 0.1
Perfume Appropriate amount Caustic potash Appropriate amount of ion-exchanged water Remaining amount
The oil layer components other than ion-exchanged water and alkali were dissolved by heating (up to 70 ° C.), neutralized with alkali, emulsified by adding ion-exchanged water, and then cooled while stirring well after emulsification.

[Example 9] Makeup base (prescription)
A W / O emulsified makeup base was prepared with the following composition.
(a) Organically modified montmorillonite 0.5% by mass
(b) Cetyl isooctanoate 2.0
(c) Octamethylcyclotetrasiloxane 2.0
(d) Decamethylcyclopentasiloxane 5.0
(e) Dimethylpolysiloxane (6cs) 5.0
(f) Liquid paraffin 3.0
(g) Dioctadecyldimethylammonium chloride 0.2
(h) Polyoxyalkylene-modified organopolysiloxane 5.0
(i) 4-t-butyl-4'-methoxydibenzoylmethane 0.3
(j) Glyceryl mono-2-ethylhexanoyl
Diparamethoxycinnamate 1.0
(k) Fine particle titanium oxide 5.0
(l) Oleyl alcohol 0.5
(m) Stearic acid 0.5
(n) Sorbitan diisostearate 4.0
(o) Antioxidant appropriate amount
(P) Perfume appropriate amount
(q) Talc 1.5
(r) Nylon powder 1.0
(s) Residual ion exchange water
(t) Sodium citrate 0.5
(u) 1,3-butylene glycol 5.0
(v) Yuzu seed 55% ethanol heated extract 0.01
(Manufacturing method)
(1) (b) to (j) and (l) to (p) are dissolved by heating, and (v) is mixed and dispersed.
(2) Add (a) to (1) and disperse and swell.
(3) Disperse (k), (q), and (r) in (2).
(4) (t) and (u) are dissolved in (s) and added to (3) to emulsify.

[Example 10] Powdery foundation (prescription)
(a) Fine particle titanium oxide 7.0% by mass
(b) Talc 40.0
(c) Mica residue
(d) Nylon powder 10.0
(e) Iron oxide red 1.0
(f) Iron oxide yellow 2.0
(g) Iron oxide black 0.2
(h) Dimethylpolysiloxane 1.0
(i) 2-ethylhexyl palmitate 9.0
(j) Sorbitan sesquioleate 1.0
(k) N, N-dimethyl PABA octyl ester 0.3
(l) Yuzu seed 60% ethanol heat extract 5.0
(m) Preservative appropriate amount
(n) Antioxidant appropriate amount
(o) Perfume appropriate amount (production method)
(1) Mix and grind (a) to (g).
(2) (h) to (k) and (m) to (o) are heated and dissolved, and (l) is mixed and dispersed.
(3) After mixing (1) and (2), mold.

[Example 11] Oily foundation (prescription)
(a) Fine particle titanium oxide 10.0% by mass
(b) Mica 22.4
(c) Kaolin 10.0
(d) Nylon powder 5.0
(e) Iron oxide red 0.5
(f) Iron oxide yellow 2.0
(g) Iron oxide black 0.1
(h) Liquid paraffin remaining
(i) Dimethylpolysiloxane 10.0
(j) Sorbitan sesquioleate 2.0
(k) Octyl methoxycinnamate 5.0
(l) Yuzu seed 90% ethanol room temperature extract 0.005
(m) Perfume appropriate amount
(n) Microcrystalline wax 6.0
(o) Carnavalou 3.0
(Manufacturing method)
(1) Mix and grind (a) to (g).
(2) (h) to (k) and (m) are dissolved by heating, and (l) is mixed and dispersed.
(3) Grind the slurry of (2) using a grinder.
(4) Add (n) and (o) to (3) and dissolve with heating, then mix and mold.

[Example 12] Cream (prescription)
Stearic acid 2.0% by mass
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 ester 2.0
Propylene glycol 5.0
Yuzu seed 40% ethanol room temperature extract 0.6
Tranexamic acid 0.2
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Propylene glycol is added to ion-exchanged water and heated to 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). Preliminarily emulsify by adding an oil phase to the aqueous phase, uniformly emulsify with a homomixer, and then cool to 30 ° C. while stirring well.

Example 13 Cream (Prescription)
Solid paraffin 5.0% by mass
Beeswax 10.0
Vaseline 15.0
Liquid paraffin 41.0
Glycerin monostearate ester 2.0
Polyethylene (20 mol)
Sorbitan monolaurate 2.0
Soap powder 0.1
Yuzu seed 30% ethanol room temperature extract 0.1
Sodium bisulfite 0.03
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue (Production method)
Add soap powder to ion-exchanged water and heat to maintain at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). The reaction is gradually added while stirring the oil phase in the aqueous phase. After completion of the reaction, the mixture is uniformly emulsified with a homomixer and cooled to 30 ° C. while stirring well after emulsification.

[Example 14] Emulsion (prescription)
Stearic acid 2.5% by mass
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
(Product name: Carbopol 94 BFGoodrich Chemical company)
Yuzu seed hot water extract 0.05
Sodium bisulfite 0.01
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchange water Residue
Dissolve the carboxyvinyl polymer in a small amount of ion-exchanged water (A phase). Polyethylene glycol 1500 and triethanolamine are added to the remaining ion-exchanged water, heated and melted, and kept at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). Add the oil phase to the water phase, preliminarily emulsify, add the A phase, uniformly emulsify with a homomixer, and cool to 30 ° C. while stirring well after emulsification.

[Example 15] Cream (prescription)
Stearic acid 5.0% by mass
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Yuzu seed 10% ethanol extract 0.1
Winged bean extract 0.1
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water Residue

[Example 16] Cream (prescription)
Liquid paraffin 3.0% by mass
Vaseline 1.0
Dimethylpolysiloxane 1.0
Stearyl alcohol 1.8
Behenyl alcohol 1.6
Glycerin 8.0
Dipropylene glycol 5.0
Yuzu seed 35% ethanol room temperature extract 0.01
Hydrolyzed silk extract 0.01
Macadamia nut oil 2.0
Hardened oil 3.0
Squalane 6.0
Stearic acid 2.0
Phytosteryl hydroxystearate 0.5
Cetyl 2-ethylhexanoate 4.0
Polyoxyethylene hydrogenated castor oil 0.5
Self-emulsifying glyceryl monostearate 3.0
Potassium hydroxide 0.15
Sodium hexametaphosphate 0.05
Trimethylglycine 2.0
Tocopherol acetate 0.1
Green tea extract 0.1
Paraben appropriate amount edetate trisodium 0.05
4-t-butyl-4′-methoxydibenzoylmethane 0.05
Diparamethoxycinnamate mono-2-ethylhexanoate glyceryl 0.05
Colorant Appropriate amount Carboxyvinyl polymer 0.05
Purified water residue

[Example 17] Cream (prescription)
Stearic acid 5.0% by mass
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Yuzu seeds 55% ethanol extract by heating 0.05
Bengle Extract 0.03
Iris Root Extract 0.02
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water Residue

[Example 18] Lotion (Prescription)
Ethanol 10.0% by mass
Glycerin 2.0
Dipropylene glycol 1.0
Isostearic acid 0.1
Poly (oxyethylene / oxypropylene) /
Methylpolysiloxane copolymer 1.0
Lauryldimethylaminoacetic acid betaine 0.1
Citric acid 0.02
Sodium citrate 0.08
Sodium hexametaphosphate 0.01
Yuzu seeds 80% ethanol extract by heating 0.3
ONONIS EXTRACT 0.1
Hipotaurine 0.1
Chamomile extract 0.1
Lavender oil 0.001
Phenoxyethanol Suitable amount Purified water Residual

Example 19 Emulsion (Prescription)
Dimethylpolysiloxane 3.0% by mass
Decamethylcyclopentasiloxane 4.0
Ethanol 5.0
Glycerin 6.0
1,3-butylene glycol 5.0
Polyoxyethylene methyl glucoside 3.0
Sunflower oil 1.0
Squalane 2.0
Potassium hydroxide 0.1
Sodium hexametaphosphate 0.05
Hydroxypropyl-β-cyclodextrin 0.1
Dipotassium glycyrrhizinate 0.05
Loquat leaf extract 0.1
Sodium L-glutamate 0.05
Fennel extract 0.1
Yuzu seed 40% ethanol room temperature extract 0.1
Thiotaurine 0.1
Yeast extract 0.1
Lavender oil 0.1
Giant extract 0.1
Dimorpholinopyridazinone 0.1
Xanthan gum 0.1
Carboxyvinyl polymer 0.1
Acrylic acid / alkyl methacrylate copolymer
(Pemulen TR-1) 0.1
Bengala Appropriate amount Yellow iron oxide Appropriate amount Paraben Appropriate amount Purified water Residual

[Example 20] Cream (prescription)
Stearic acid 5.0% by mass
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Yuzu seeds 80% ethanol heated extract 0.1
Cuckoo extract 0.1
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water

[Example 21] Lotion (Prescription)
Ethyl alcohol 5.0% by mass
Glycerin 1.0
1,3-butylene glycol 5.0
Polyoxyethylene polyoxypropylene
Decyl tetradecyl ether 0.2
Sodium hexametaphosphate 0.03
Trimethylglycine 1.0
Sodium polyaspartate 0.1
Thiotaurine 0.1
Green tea extract 0.1
Western mint extract 0.1
Turmeric extract 0.1
Yuzu seed 99% ethanol room temperature extract 0.01
Lysolecithin 0.01
EDTA 3 sodium 0.1
Carboxyvinyl polymer 0.05
Potassium hydroxide 0.02
Phenoxyethanol Appropriate amount Purified water Residual perfume Appropriate amount

[Example 22] Cream (prescription)
Stearic acid 5.0% by mass
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Yuzu seeds 95% ethanol heated extract 0.01
Mishima psycho extract 0.01
Time extract 0.01
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water

[Example 23] Cream (prescription)
Liquid paraffin 8.0% by mass
Vaseline 3.0
Dimethylpolysiloxane 2.0
Stearyl alcohol 3.0
Behenyl alcohol 2.0
Glycerin 5.0
Dipropylene glycol 4.0
Trehalose 1.0
Yuzu seed 85% ethanol heat extract 0.01
Asenya extract 0.01
Tetra-2-ethylhexanoic acid pentaerythrit 4.0
Polyisoethylene glyceryl monoisostearate 2.0
Polyoxyethylene glycerol monostearate 1.0
Lipophilic glyceryl monostearate 2.0
Citric acid 0.05
Sodium citrate 0.05
Potassium hydroxide 0.015
Oil-soluble licorice extract 0.1
Retinol 0.25
Tocopherol acetate 0.1
P-Hydroxybenzoate appropriate amount phenoxyethanol appropriate amount dibutylhydroxytoluene appropriate amount edetate trisodium 0.05
4-t-butyl-4′-methoxydibenzoylmethane 0.01
2-Ethylhexyl paramethoxycinnamate 0.1
β-carotene 0.01
Polyvinyl alcohol 0.5
Hydroxyethyl cellulose 0.5
Carboxyvinyl polymer 0.05
Purified water Residual fragrance Appropriate amount

[Example 24] Cream (prescription)
Decamethylcyclopentasiloxane 15.0% by mass
Trimethylsiloxysilicate 5.0
Polyoxyethylene / methylpolysiloxane copolymer 5.0
Glycerin 5.0
1,3-butylene glycol 5.0
Maltitol solution 2.0
Macadamia nut oil 2.0
Squalane 2.0
Phytosteryl hydroxystearate 0.5
Cetyl 2-ethylhexanoate 2.0
Distearyldimethylammonium chloride 0.2
Retinol acetate 0.02
Tocopherol acetate 0.05
Fish collagen 0.4
Sodium chondroitin sulfate 0.01
Sodium hyaluronate 0.1
Yuzu seed 45% ethanol room temperature extract 0.01
Edetate trisodium 0.05
Diparamethoxycinnamic acid mono-2-
Glyceryl ethylhexanoate 0.05
Aluminum magnesium silicate 0.3
Paraben Appropriate amount of purified water Residual

[Example 25] Cream (prescription)
Stearic acid 5.0% by mass
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Yuzu seed 20% ethanol heat extract 0.02
Marjoram extract 0.1
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water

[Example 26] Cream (prescription)
Stearic acid 5.0% by mass
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Yuzu seed 10% ethanol heat extract 0.01
Tormentilla extract 0.01
Clara extract 0.01
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water

[Example 27] Emulsion (prescription)
Dimethylpolysiloxane 2.0% by mass
Decamethylcyclopentasiloxane 25.0
Dodecamethylcyclohexasiloxane 10.0
Polyoxyethylene / methylpolysiloxane copolymer 1.5
Trimethylsiloxysilicic acid 1.0
1,3-butylene glycol 5.0
Yuzu seed 75% ethanol room temperature extract 0.01
Mangosteen extract 0.01
Squalane 0.5
Talc 5.0
Dipotassium glycyrrhizinate 0.1
Tocopherol acetate 0.1
Edetate trisodium 0.05
4-t-butyl-4′-methoxydibenzoylmethane 1.0
2-Ethylhexyl paramethoxycinnamate 5.0
Diparamethoxycinnamic acid mono-2-
Glyceryl ethylhexanoate 1.0
Silicone coated fine particle titanium oxide (40nm) 4.0
Dimethyl distearyl ammonium hectorite 0.5
Spherical polyethylene powder 3.0
Phenoxyethanol Appropriate amount Purified water Residual perfume Appropriate amount

[Example 28] Bath preparation (prescription)
Squalane 10.0% by mass
Macadamia nut oil 10.0
Sorbitan oleate 5.0
POE oleyl ether 10.0
Yuzu Seed Extract 0.5
Fennel extract 0.5
Juyaku Extract 0.5
Ion exchange water 0.5
Dye Appropriate amount of perfume Appropriate amount

[Example 29] Lotion (Prescription)
1,3-butylene glycol 9.0% by mass
Dipropylene glycol 3.0
Glycerin 5.0
Ethanol 3.0
Trimethylglycine 5.0
PEG / PPG-14 / 7 dimethyl ether 2.0
PEG-60 hydrogenated castor oil 0.7
Yuzu seed extract 0.3
Yuzu (fruit) extract 0.3
Citric acid 0.02
Sodium citrate 0.09
Sodium metaphosphate 0.1
Methyl paraben Appropriate amount Phenoxyethanol Appropriate amount Deionized water Residual perfume Appropriate amount

Claims (3)

A type IV collagen production promoter comprising a solvent extract of yuzu seeds , wherein the solvent is hydrous ethanol . A type VII collagen production promoter comprising a solvent extract of yuzu seeds , wherein the solvent is hydrous ethanol . A laminin-5 production promoter comprising a solvent extract of Yuzu seeds , wherein the solvent is hydrous ethanol .