TW200808366A - Wrinkle preventive/remedy - Google Patents

Abstract

To provide a safe wrinkle preventive/remedy with comprehensive wrinkle preventing/remedying effect that through inhibition of skin hyperplasia induced at the time of dropping of barrier function due to drying, etc., attains the prevention and remedy of wrinkles, especially fine wrinkles, attributed to the barrier function dropping and simultaneously attains the prevention and remedy of wrinkles attributed to aging and photo-aging. There is provided a wrinkle preventive/remedy comprising as an active ingredient at least one compound selected from the group consisting of specified aminosulfuric acid compounds and salts thereof.

Description

。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 More specifically, it is to prevent skin thickening caused by a decrease in defense function due to drying, etc., to prevent and improve wrinkles, especially small wrinkles, which are caused by such a defense function, and further, to provide prevention and improvement due to age increase or A wrinkle-free and improver that is a comprehensive anti-wrinkle and anti-wrinkle effect caused by photoaging. [Prior Art] The stratum corneum located at the outermost layer of the skin has contact or penetration of foreign matter such as bacteria or harmful substances, protects the living body, and maintains the defensive function of the skin in a healthy state by preventing evaporation of water from the body. Excessive contact with water or dryness in winter, excessive cold weather in summer, etc., and reduced defensive function, increased trails epidaermal water loss (TEWL) from the surface of the skin, relative keratinous water content reduce. When the texture (texture) formed by the skin groove or the ridge is irregular due to the decrease in the water content of the stratum corneum, the skin is rough and rough (see Non-Patent Document 1). In the report, it is pointed out that the condition of small wrinkles is such that when the skin is rough, the irregular skin lines are formed in a certain direction. In fact, the skin of the stratum corneum has a small water content, and the area ratio of small wrinkles is high ( Refer to Non-Patent Document 2). Generally, the dry state continues to promote the formation of small wrinkles, but the influence of the season, modern women due to the popularity of air-conditioners, that is, -5 - 200808366 (2) so that summer skin is still often in a dry environment, between the ages of 25 and 29 There is an increasing trend between the worries of small wrinkles. In order to prevent this skin defense function from being reduced, it is proposed to prevent or improve the moisture evapotranspiration from the skin surface, followed by rough skin or small wrinkles, coating glycerin or natural moisturizing factor NMF (related components), collagen derivatives A method of moisturizing the skin, improving the moisture content of the skin, using a cell-revitalizing component such as Placenta extract or vitamin φ, and promoting the keratinocyte regeneration, but the current situation is consistent with The formation mechanism of small wrinkles, and the agent having the excellent wrinkle improving effect above the moisturizing effect is almost completely undeveloped. On the other hand, wrinkles also occur due to aging, and the wrinkles that occur with aging are particularly high in middle-aged ages, among which women are extremely distressed. The demand for the improvement of wrinkles which occurs with this aging is very high, but because of the mechanism of aging and wrinkles, there are still many unclear parts. In the conventional cosmetics, efforts are made to improve the wrinkles as described above. The method of maintaining water, etc. • The current situation has not been able to achieve sufficient results as described above. As a result of the wrinkles which occur with aging, only the conventional method can not adequately prevent skin aging and wrinkles from occurring. In addition, in recent years, with the development of aging, as a cause of skin aging, as a whole, an important factor in the ageing system, and further, it is found that oxidation, sunlight (ultraviolet rays), drying as described above, etc. are also related to the skin. The direct factor of aging. Among these factors, especially sunlight (ultraviolet light), especially in the change called photoaging, is becoming more and more obvious. The whole part of the body is the part that is most easily photoaging, and the skin of photoaging is in the skin. 6- (3) (3) 200608366 The most important matrix component of collagen fibers is significantly reduced. Next, it also shows that the occurrence of wrinkles and the disappearance of skin tension are closely related to the reduction of collagen fibers. As for the occurrence of wrinkles and skin aging, with various skin aging factors, among which, especially exposure to sunlight (ultraviolet rays), the proliferative activity of fibroblasts of major cells in the dermis or the synthesis function of collagen and the like are reduced, such collagen, etc. The regeneration speed is also slower. As a result of the deterioration of the skin elasticity, wrinkles, and skin aging, it is known that a substance having an effect of improving such wrinkles is known to be effective as an amino acid alanine and glycine (refer to Patent Document 1 and Patent Document 2). . However, when such alanine or glycine is applied to the skin, it is accompanied by abnormal pain and difficulty in use, and at this stage, there is no such high safety and a good wrinkle improving agent. In addition, furthermore, various factors such as wrinkles and skin aging, such as the above-mentioned drying or photoaging, are complicatedly interlaced, since it is not sufficient to try to prevent or improve the crepe by an effect, so comprehensive improvement is required. A wrinkle effect and an agent that inhibits skin aging. The N-(2-acetamido)-2-aminoethanesulfonic acid of the present invention is known as a constitutive compound of Good's buffer (refer to Non-Patent Document 3) used biologically. In addition, it is known that it is used for the purpose of reducing pores (refer to Patent Document 3) and for preventing dryness of skin contained in a moisturizing agent (refer to Patent Document 4), but N-(2-acetamido)-2- is not known. Aminoethane sulfonic acid has the effect of preventing and improving wrinkles. Patent Document 1: Japanese Patent Publication No. Hei 1 1 -49628

Patent Document 2: Japanese Unexamined Patent Publication No. Hei No. Hei No. Hei No. Hei No. 2, No. 2 0 0 5 -1 7 9 3 4 No. 3 Patent Publication No. 4: W002-083136 Etc., Fragrance and Makeup, 丨 5, 2, 60 to 65, 1991 Non-Patent Document 2: Sui Chuan, et al., Frangrance Journal, 1 992 (1 1 ), p.29-42

Non-Patent Document 3: E. Good, Biochemistry, 5(2), p, 467, 1966 SUMMARY OF THE INVENTION PROBLEM TO BE SOLVED BY THE INVENTION The present invention has been made in view of the above circumstances, and the object is to provide suppression. Skin thickening caused by a decrease in defense function due to drying, etc., prevention and improvement of wrinkles, such as small wrinkles, which are caused by such defensive functions, and further provide "prevention and improvement of wrinkles caused by aging or photoaging" A comprehensive anti-wrinkle • a wrinkle-free and improver that improves the wrinkle effect. In order to solve the above problems, the inventors of the present invention have found that N-(2-ethylamino)-2-aminoethanesulfonic acid and the salt prevent and improve the drying caused by drying. Further, the present invention has been completed in order to prevent and improve the anti-wrinkle/improvement effect and the safety of the wrinkles caused by light aging -8-(5) 200808366. In other words, the present invention contains an anti-wrinkle/improving agent containing two or more kinds of an amine-based sulfuric acid compound represented by the following formula (1) and a compound in which the salt is a group. [化2] 0

Ch2_nh—ch2-c

II

-OH Ο Ο) EFFECT OF THE INVENTION According to the present invention, an excellent wrinkle-reducing agent having an anti-wrinkle and improvement effect can be obtained. BEST MODE FOR CARRYING OUT THE INVENTION The following is a detailed description of the best mode for carrying out the invention. In the present invention, one or two or more kinds of compounds selected from the group consisting of the aminosulfuric acid compounds represented by the following formula (1) and the salts thereof are used. -9 - 200808366 (6) [Chemical 3] Ο

CH:—ΝΗ—CHj-CHa—S OH

II ο (1) The aminosulfuric acid compound represented by the above formula (1) used in the present invention is N-(2-acetamido)-2-aminoethanesulfonic acid, N-aminoformamidine. A chemical compound of methyl taurine or 2-[(2-amino-2-oxoethyl)amino]ethanesulfonic acid, etc., generally known as the compound of the name ACES. Hereinafter, in the present invention, the aminosulfuric acid compound represented by the above formula (i) is referred to as ACES. The ACES used in the present invention can be synthesized by a known method, and a commercially available product such as Aldrich Co., Ltd. is also used. The above-mentioned ACES salt is not particularly limited, and examples thereof include hydrochlorides, sulfates, phosphates, hydrobromides, sodium salts, potassium salts, magnesium salts, calcium salts, and ammonium salts of inorganic salts. The organic salt may, for example, be an acetate, a lactate, a maleate, a fumarate, a tartrate, a citrate, a methanesulfonate, a p-toluenesulfonate, a triethanolamine salt, a diethanolamine salt or an amine. Acid salt and the like. Salts can be obtained by known methods. Any of the ACES and the salt system of the present invention has an excellent anti-wrinkle/improvement effect as will be described later. Therefore, one or two or more compounds selected from the group consisting of ACES and the salt are contained as an active ingredient to prepare an anti-wrinkle/improving agent. The above-mentioned wrinkle-improving agent is based on the novel and effective use of the above-mentioned novel functions of the above-mentioned inventive ACES and -10- 200808366 (7) salts. In the anti-wrinkle/improving agent, one or two or more compounds selected from the group consisting of ACES and the salt are contained in an amount effective to prevent wrinkles and improve the amount of the wrinkle-reducing agent. 0. 〇〇1 to 2 0.0% by mass is preferred. Further, it is preferably 0.1 to 1 〇. The anti-wrinkle/improving agent of the present invention can be produced by a usual method, and as a constituent of the anti-wrinkle/improving agent, one or two or more types of compounds selected from the group consisting of aces and the salt can be prepared. However, it is usually not limited to the extent of the efficacy of the present invention, and other components used for external preparations for skin such as cosmetics or pharmaceuticals containing the external products of the medical department can be appropriately blended as needed. Examples of the other components (optional components) include an oil component, a surfactant, a powder, a color material, water, an alcohol, a tackifier, a chelating agent, a polyoxyalkylene, an antioxidant, an ultraviolet absorber, and a moisturizing agent. Agents, perfumes, various medicinal ingredients, preservatives, pH adjusters, neutralizers, etc. The anti-wrinkle/improving agent described above is used in the form of a shape such as a cosmetic or a medical composition, and exhibits an excellent anti-wrinkle and improvement effect. In particular, it is suitably used in the form of a skin external preparation for external use on the skin, and is applied by a method of directly applying or spreading to the skin, and exhibits an excellent anti-wrinkle/improvement effect. The anti-wrinkle/improving agent of the present invention is suitable for use on the skin, and can be used for a cosmetic method for preventing wrinkles from forming and/or reducing and eliminating wrinkles. The usage and amount of the anti-wrinkle/improving agent of the present invention in the related cosmetic method are not particularly limited, and may be appropriately determined depending on the dosage form or the wrinkle state of the skin, but it is usually several times per day, for example, once. Up to 5 times -11 - 200808366 (8), appropriate amount, for example, 0.1 ml to 1 ml per lcm2, directly rubbed into the skin or soaked in the appropriate amount of gauze, etc., attached to the skin. The dosage form of the anti-wrinkle improving agent is not particularly limited as long as it can exert the effects of the present invention, and examples thereof include a solution system, a solubilization system, an emulsification system, a powder dispersion system, a water-oil two-layer system, and a water/oil. - Any of the three forms of powder, ointment, gel, aerosol, and the like. The anti-wrinkle/improving agent can be applied to, for example, the field of cosmetics and pharmaceuticals including the external portion φ of the medical department, and examples of the product include ointments, creams, lotions, lotions, essences, jellies, gels, and the like. Any form such as a pack, a mask, or a cosmetic. [Embodiment] The present invention will be more specifically described by way of Examples and the like, but the technical scope of the present invention is not limited to the embodiments. The blending amount is % by mass unless otherwise specified. [Prevention of Wrinkle Mode Using Destructive Defense • Effect of Improving Small Wrinkles] Nude mice (HR-1, male 6 weeks old, Hoshino experimental animals) were divided into 2 groups of 7 each, on the left back of nude mice, 4 to Tape stripping at 8 mg/cm2/h, 3 times a week for 4 weeks, to evaluate the inhibition of wrinkles. Each time the tape was peeled off, each of the compositions of Example 1 (with A C S E ) or Comparative Example (without combination) shown below was applied. -12- 200808366

[Example 1] Composition

ACES Cosmetics Ethanol Refined Water Blend (% by mass) 1.0 50.0 Remaining

[Comparative Example 1] Composition Ingredients Cosmetic Ethanol Refined Water Blend amount (% by mass) 5 0.0 Remaining The wrinkle occurrence after 4 weeks of visual observation by four judges was scored. The occurrence of wrinkles was evaluated according to the criteria shown in Table 1 below, and scored on a scale of 〇 · 5 . The greater the score, the deeper the wrinkles. The average enthalpy and standard deviation of each group were calculated, and the effects of each composition are shown in Fig. 1. In Fig. 1, "ACES" indicates the results of the coating group of Example 1 (combined with ACSE), and "Vehicle" indicates the results of the coating group of Comparative Example 1 (without the drug). It is confirmed from Fig. 1 that the first embodiment of the ACSE according to the present invention exhibits an intentional wrinkle suppressing effect as compared with the vehicle. -13- 200808366 (10) [Table 1] Rating: Judgment criteria 〇 : No wrinkles are considered. 1 : The wrinkles are light and light. 2: Wrinkles are obvious. 3: Deep wrinkles. Next, the skin thickness of the back of the nude mice subjected to the above treatment was measured, and three portions of each portion were measured using a digital thickness gauge (Peacock) to determine an average enthalpy. It is known that the degree of wrinkle formation and skin thickness are correlated, and the skin thickness is reduced, indicating that wrinkle formation is suppressed. The results of skin thickness are shown in Figure 2. In Fig. 2, "ACES" indicates the results of the coating group of Example 1 (with the ACSE), and "Vehicle" indicates the results of the coating group of Comparative Example 1 (without the drug). In addition, in Fig. 2, the vertical axis indicates TS/NT, and the thickness ratio of the TS on the left side of the back of each nude mouse to the NT on the right side (control group) is calculated, and the increase rate of the left and right skin thickness is calculated, and the average of each group is obtained. value. The larger the TS/NT ratio, the greater the difference in skin thickness between the left and right, and the deeper wrinkles. As shown in Fig. 2, Example 1 in combination with the ACES of the present invention significantly inhibited skin thickness as compared with Vehicle. From the results, it was confirmed that Example 1 of the ACES incorporating the present invention inhibited skin hypertrophy to prevent wrinkle formation. [Photon Aging Wrinkle Improvement Test in Nude Mice] Nude mice were treated with Hr-Ι (Skh-Ι) nude mice (Hoshino experimental animals; 6 weeks to 10 weeks old), and some methods of Schwartz et al. were changed ( -14- 200808366) (11)

Haratake A. et al. J. Invest. Dermatol. 108: 769-775, 1997), based on the method of repeatedly irradiating UVB (Naganuma M. et al. J. Dermatol. Sci. 25:29-3 5, 200 1? Schwartz EJ Invest.

Dermatol 91: 158-161, 1 988), causing wrinkles. That is, the back side is irradiated with UVB (light source; Toshiba Electric, Toshiba FL-20SE fluorescent lamp), 3 times a week for 10 weeks. The amount of irradiation after the start was 36 mJ/cm 2 /time, and gradually increased after the second week, and the first week was 216 mJ/cm 2 /time. The total exposure was 4.6 J/cm2. The amount of ultraviolet light was measured using UVRADIOMETER (UVR-305/3 65D ( Π ) , Topcon). After the end of the ultraviolet irradiation, photographs of the back of the nude mouse were taken to change the method of Bissett et al. (Bissett DL. et al. Photochemistry and Photobiology 46: 367-3 78, 1 987), based on the criteria shown in Table 2 below. The degree of occurrence of the scored wrinkles was applied only to nude mice having a wrinkle score of 7 or more, and the above Example 1 (combined with ACES) or Comparative Example 1 (uncompounded with a drug) was applied. The wrinkle scoring operation was performed by three measurers, and the score was determined by discussion. -15- 200808366 (12) [Table 2] Rating: Judgment criteria 〇 · Consider countless lines. 1 : Shallow, short, or small in number 2. 2: Think of light and wrinkles. 3: Deeper or longer than the score 2. 4 than the score 4: I think there are light and light wrinkles on the whole surface. 5: Deeper than the score of 4, or longer. The ratio is 6 6 : It is considered to have deep and long wrinkles. 7: Increases wrinkles deeper and longer than score 6. 8: I think there is deep and long wrinkles on the entire surface. The nude mice with a score of 7 or more were divided into two groups and the scores were the same. The nude mice in each group were treated with a back skin of 100 μl as shown in the above composition once a week for 5 times for 6 weeks. Instead of the example 2 (co-rotating right-handed-α-fertility), \)j HEPES: Ν-[2-ethyl]piperazine-oxime, -[2·B. [Comparative Example 2] Composition Ingredients Formulation L-dextrorotatory-α-tocopherol Cosmetic ethanol Refined water, short, or small in quantity. I, or short. It is lighter or shorter than the score of 8. The same amount (% by mass) was carried out by grouping each of the two skins, and the two skins as a whole, continuously coating each or Example 1, and comparing it to the following as shown below: and replacing it with Comparative Example 3 (sulphonic acid). 3.0 50.0 Remaining 16- 200808366 (13) [Comparative Example 3] Composition amount (% by mass) 3.0 50.0 Residual ingredients

After the application of HEPES cosmetic ethanol refined water, photographs of the back of the nude mouse were taken. According to the criteria shown in Table 2 above, the three animal subjects were scored according to the state of the Tibetan animal group, and the score was determined by discussion. . The improvement of wrinkles in nude mice was determined as follows. That is, "wrinkle improvement degree" = "score before coating solution" - "score after coating solution for 6 weeks" is calculated. Fig. 3 (ACES), Fig. 4 (left-handed spin-α-tocopherol), and Fig. 5 (HEPES) show the wrinkle improvement degree (wrinkle improvement change amount) obtained according to the above formula. In addition, in Fig. 3, F is Example 1 (combined with ACES), and X is Comparative Example 1 (unfilled with a drug). Further, in Fig. 4, B is Comparative Example 2 (coordinated with L-dextrorotatory-α-tocopherol), and X is compared with Example 1 (uncompounded with a drug). In addition, in Fig. 5, Ε is Comparative Example 3 (with HEPES), and V is Comparative Example 1 (without a drug). Further, in Fig. 5, the same symbol as that of Fig. 4 is used for the data symbol, and the amount of wrinkle improvement change 1 in E is overlapped by 4 points in E, and the wrinkle improvement change amount 1 in V is overlapped by 3 points. As shown in Fig. 3, with respect to Comparative Example 1 (uncompounded drug) coating group -17-200808366 (14) (mean score: 1.8), Example 1 (ACES) coating group of the present invention (average score: 3.0), significantly promotes the reduction of wrinkles. From the above, it was confirmed that the ACES solution has an effect of improving skin wrinkles due to ultraviolet rays. On the other hand, as shown in Fig. 4, the left anti-oxidation substance L-dextrorotatory-α-tocopherol was compared with the comparative example 1 (uncompounded drug) coating group (average score: 3 · 0), Comparative Example 2 (Left-dextrorotatory-α-tocopherol) was coated with φ cloth group (average score: 3.2), and it was considered that especially the number or formation of wrinkles was not different, and wrinkles were not observed at all. As a result of the improvement of skin wrinkles formed by ultraviolet rays, it is considered that the antioxidant substances have no particular effect. In addition, as shown in FIG. 5, the HEPES which is the most suitable in the patent document 4 which contains the effect of containing a moisturizing agent to prevent the skin from drying is compared with the coating group of the comparative example 1 (uncompounded agent) (average rating: 1 · 0 In the comparative example 3 (HEPES) coating group (average score: 1.4), it was considered that especially the number or formation of wrinkles was not different, and wrinkle reduction was hardly observed at all. The improvement in skin wrinkles formed by the outer line of purple Φ is not related to the effects described in Patent Document 4. [Human eye wrinkle improvement test] (Method) In the face of a healthy male assessor with more wrinkles, a half face method (blind test) was applied to coat Example 2 as follows. And the lotion of Comparative Example 4, 3 times a day. Example 2 Lotion -18- (15) 200808366 Ingredients Ingredients (% by mass) ACES 2.0 Ethanol for Cosmetics 15.0

If water is used in the remaining part of the sample 4, the amount of the cosmetic water is adjusted by the amount of the refined water (% by mass). 15.0 The remaining SILFLO (Flexico Development Ltd.) is used, and the coated portion is taken from the end of the eye to obtain a coating. The rate of change in wrinkle area ratio before (〇M) and after 1 month of application (1M) and after 2 months of coating (2M). The wrinkle area ratio is obtained by analyzing a replica sample by a wrinkle measuring device by a laser light cutting method (refer to Japanese Laid-Open Patent Publication No. Hei 7- 1 1 3 6 2 3). #About the area ratio of wrinkles before coating and after 1 month of coating (1Μ), 2 months after coating (2 Μ), and the rate of change (%, vsOM) at 1% before coating. The average 値 is shown in Figure 6. In the comparative example 4, the wrinkle area of the lotion-coated side (control group) hardly changed before the application, and the wrinkle area of the lotion-coated side (ACES) of Example 2 was reduced by 80% or less compared with that before the application. After 2 months, the comparison between the lotion water of Comparative Example 4 and the lotion of Example 2 was considered to have a trend difference (student t test, Student, s attest). From this, the effect of the wrinkle improvement of the lotion of Example 2 was confirmed. -19- 200808366 (16) [Promoting laminin5 (fiductin 5) production effect test] (1) Culture epidermal keratinocyte epidermal keratinocyte cell line isolated from human foreskin, cultured in epidermal cell proliferation medium (KGM) with low calcium concentration . The bovine pituitary extract and EGF (Epidermal Growth Factor) were added to the medium. After the cell line was cultured to the 4th passage with KGM, it was treated with trypsin-EDTA to float the conjugated cells, and the cells were removed by filtration to obtain a uniform cell suspension. The cells were collected by centrifugation and resuspended in DMEM (Dulbecco 5s Modification Eaglet Medium) -F12 (2:1) -0.1% BSA to 8xl04/ml. 0.5 ml of this cell suspension was added to the same medium containing 2.5 ml of a 2-fold concentration of the drug. The culture system was carried out at 37 ° C for 24 hours using a 24-well culture plate. At the end of the culture, the culture supernatant was transferred to a microcentrifuge tube, centrifuged at 100 rpm for 5 minutes, and the supernatant was transferred to a new microcentrifuge tube and stored at 20 ° C until the measurement day of the adhesion protein 5. Further, in the solubilized cells and the fibronectin 5 bound to the cultured plastic, a Tris-hydrochloric acid buffer solution (pH 7.4) containing various surfactants was added to each well, and stored at -20 ° C overnight. The next day, ultrasonic processing was performed and frozen again. The next day, after dissolving again, the mixture was centrifuged at 1000 rpm for 5 minutes, and the supernatant was transferred to a microcentrifuge tube and stored at -2 °C until the measurement day of the fibronectin 5. (2) Determination of adhesion protein 5 by using the sandwich ELISA method. -20- 200808366 (17) The fibronectin 5 line present in the culture supernatant and cell layer was determined by sandwich enzyme immunoassay. On the fixed layer of the 96-well ELISA plate, the monoclonal antibody against the α3 chain of the adhesion protein 5, BM 165 was ligated. In order to sandwich the adhesion protein 5 for measurement, as another anti-system, 6F 12 biotinylated (b-6F 1 2 ) with respect to the monoclonal antibody of the triple chain of the adhesion protein was used in advance. In this method, only the heterotrimer (α 3 ^ 3 r 2 ) which can function is measured, and the heterodimer (yS 3 r 2 ) is not detected. The sample was added to the pre-φ before being placed in each well of a 3% pectin•phosphate buffer solution containing b-6F12. The final dilution rate of the sample in the well was 1/4 of the culture medium and the cell layer was 1 /1 〇. The antigen-antibody reaction was carried out at 37 ° C for 2 hours. After washing the culture plate, a solution of avidin-horseradish peroxidase (avidin-HRP) was added, and the reaction was further carried out for 30 to 1 hour. After washing, a solution of HRP matrix was added to a solution of <2>2丨11〇-bis (3-ethylbenzthiaoline-6-sulfonic acid), and the absorbance at 4,05 nm was measured by an enzyme immunoassay (ELISA plate reader). The calibration curve was made in the range of 0 to 40 ng/ml. (3) The amount of production of fibronectin 5 The amount of production of fibronectin 5 is the sum of the amount of free medium in the medium and the residual amount of the cell layer, which is expressed as the relative enthalpy of the sample (control group) to which no plant extract is added. . The results are shown in Table 3. As is apparent from Table 3, although the creatinine which is the same glycine analog also has an effect of promoting the production of fibronectin 5, the ACES system has a higher promoting effect. -21 - 200808366 (18) [Table 3] Sample concentration (%) Relative to the amount of recombinant protein 5 produced without added control (%) ACES 0.0001 124 0.001 137 Creatine 0.001 111 [Type IV collagen, VE Collagen-promoting effect test] (1) Culture of human fibroblasts Human fibroblasts cultured in DMEM medium containing 10% FBS were inoculated into a 24-well culture dish, and after the cells were attached, they were changed to contain 0.25% FBS and 250// D ME Μ medium of ascorbyl glucoside, add the drug. One day later, the culture supernatant was recovered, centrifuged, and the type IV collagen, W collagen, and the amount of DNA in the supernatant were measured to obtain an index of the number of cells. (2) DNA quantification The amount of DNA was measured by the fluorescence measurement method of Hoechst Co., Ltd. 3 3 3 42. (3) Determination of Type IV and VD Collagen Type IV and W Collagen by sandwich enzyme immunoassay (Sandwich ELISA) was determined by sandwich enzyme immunoassay. The antibodies used in this example are shown below.

• Type IV collagen-specific antibody; monoclonal antibody JK-199 and multiple antibodies MO_S-CLIV -22- 200808366 (19) • νπ-type collagen-specific antibody; monoclonal antibody Np-185 and monoclonal antibody NP- 32 The amount of type 1V and type I collagen in the DNA of the drug-added sample when the amount of type IV and W-type collagen in the DNA of the sample without the drug (control group) was 1 ,, as the type IV, Fox-type collagen production. The results are shown in Table 4. As is clear from Table 4, ACES has an effect of promoting the production of type IV collagen and apical collagen. [Table 4] Sample concentration (%) Relative type IV collagen production amount (%) relative to no added control group Relative relative type VII collagen production amount (%) with no added control group ACES 0.0001 108 103 0.001 113 106 [Artificial skin fibronectin 5, type IV collagen, fox-type collagen staining test] Collagen gel is made on fiber from human dermal fibroblasts (1 X 1 05 ce 11 s / m 1 ) After suspending the collagen solution, the collagen was gelatinized at 37 ° C in a 60 mm Petri dish. Thereafter, the gel was peeled off from the wall of the dish, a collagen gel was placed on the metal, and a glass ring (inner diameter of 12 mm) was placed on the gel. A mixed medium containing a suspension of keratinocytes from the human foreskin was added, but no liquid was permeated into the glass ring. The epidermal cells were adhered overnight and the glass rings were removed the next day. The above-mentioned culture medium was filled to the boundary layer of the epidermis, and the skin layer was exposed to the air to prepare a skin pattern having a re-layered epidermis having a stratum corneum layer formed by -23-(20) 200808366. After the fourth day of inoculation of epidermal cells, the medium was replaced with a medium containing (1) unadded drug, (2) 1% ACES, (3) 2% ACES, (4) 2% ACES + 1% creatinine. Then, the medium containing the same kind and the same concentration was replaced every 2 to 3 days, and cultured for another 2 weeks. The artificial skin formed was stained with hematoxylin-eosin staining and immunochromatography (anti-adhesin 5 antibody, anti-type IV collagen antibody, and VD collagen antibody). The staining of fibronectin type 5, type IV and type W collagen was graded from low to high in five stages from 1 to 5. The results are shown in Table 5. Table 5 The staining degree of fibronectin 5 The staining degree of type IV collagen The staining degree of type VII collagen 0) 2 2 1 (2) 3 3 2 (3) 4 5 5 (4) 5 5 5

From the control (1) of Table 5, the fibronectin type 5 and type IV collagen were slightly stained, and the VD type collagen was hardly observed. On the other hand, the ACES system which promotes the action of collagen 5·IV collagen-type 胶原-type collagen is involved in any of these dyeing properties, and further, by the action of the muscarin having an action of adhesion protein 5. The acid combination improves the dyeability of the adhesion protein, which is also confirmed in artificial skin. The skin external preparation which is an example of the preparation of the anti-wrinkle/improving agent of the present invention is shown below. In addition, any skin external preparation has excellent anti-24- (21) 200808366 wrinkle and improvement effect. Formulation Example 1: Anti-wrinkle and improving cream Ingredients Amount (% by mass) Stearic acid 5.0 Stearyl alcohol 4.0 Isopropyl myristate 18.0

Glycerol monostearate 3.0 10.0 propylene glycol

ACES Caustic Potassium Sodium Bisulfite Preservatives Perfume Ion exchange water (preparation) Add propylene glycol Heat at 7 °C (aqueous phase) at 7 °C (oil phase). After slowly, temporarily maintain the temperature, and then fully agitate and cool to 2 0.0 0.2 0.01. Appropriate amount The remaining amount and caustic potash are dissolved in ion-exchanged water. Mix other ingredients, heat to dissolve, and add the oil phase to the water phase. Thereafter, it was produced by uniformly homogenizing the emulsion at 30 °C. -25- (22) (22)200808366 Formulation Example 2: Anti-wrinkle • Improved cream ingredients (% by mass) Stearic acid 6.0 Mountain, sorbitan monostearate 2.0 Polyethylene oxide ( 20 mol) sorbitan monostearate 1.5 propylene glycol 10.0 ACES 7.0 glyceryl tricaprylate 10.0 squalene 5.0 sodium bisulfite 0.01 p-ethyl benzoate (Ethylparab en) 0.3 perfume amount of ions The exchange water remaining (preparation method) was added to propylene glycol to dissolve in ion-exchanged water, and the heating was maintained at 70 ° C (aqueous phase). Mix other ingredients, heat to dissolve, and keep at 7 ° C (oil phase). The oil phase was added to the aqueous phase to carry out preliminary emulsification, and the mixture was uniformly emulsified by a homogenizer, and then uniformly stirred and cooled to 30 ° C to be produced. -26- (23) (23)200808366 Formulation Example 3: Anti-wrinkle • Improved cream ingredients (% by mass) Stearyl alcohol 7.0 Stearic acid 2.0 Hydrogenated lanolin 2.0 Squalane 5.0 2 - Octyl twelve Alcohol 6.0 Polyethylene oxide (25 mol) Cetyl ether 3.0 Glycerol monostearate 2.0 Propylene glycol 5.0 ACES 0.001 Perfume amount of sodium bisulfite 0.03 Ethyl p-hydroxybenzoate 0.3 Ion exchange water remaining (preparation method The propylene glycol was added to dissolve in ion-exchanged water, and the heating was maintained at 7 ° C (aqueous phase). Mix other ingredients, heat to dissolve, and keep at 70 °C (oil phase). The oil phase was added to the aqueous phase to carry out preliminary emulsification, and the mixture was uniformly emulsified by a homogenizer, and then uniformly stirred and cooled to 30 ° C to be produced. -27- 200808366 (24) Formulation Example 4: Anti-wrinkle • Improved amount of cream ingredients (% by mass) Stearic acid 2.5 Cetyl alcohol 1.5 5.0 10.0 Vaseline mobile paraffin polyethylene oxide (1 0 mol ) monooleate 2.0

Polyethylene glycol 1 5 00 3.0 Triethanolamine 1.0 ACES 10.0 Sodium bisulfite 0.01 Ethyl p-hydroxybenzoate 0.3 Carboxyvinyl polymer 0.05 Perfume amount of ion exchange water Residue (preparation method) Dissolve carboxyvinyl polymer in a small amount of ions Exchange water (phase A). Polyethylene glycol 1 500 and triethanolamine were added to the remaining ion exchange water, and dissolved by heating to maintain at 70 ° C (aqueous phase). Mix other ingredients, heat to dissolve, and keep at 70 ° C (oil phase). The oil phase was added to the aqueous phase to carry out preliminary emulsification, and the phase A was added, and the mixture was uniformly emulsified by a homogenizer, and the mixture was emulsified in a sufficient amount, and then stirred and mixed, and cooled to 30 ° C to be produced. -28- 200808366 (25) Formulation Example 5: Anti-wrinkle and improving emulsion Ingredients Ingredients (% by mass) (oil phase) Stearyl alcohol 1.5 Squalene 2.0 Vaseline 2.5 Deodorized liquid lanolin 1.5

Evening Primrose Oil 2.0 Isopropyl myristate 5.0 Glycerol monooleate 2.0 Polyethylene oxide (60 mol) Hardened castor oil ester 2.0 Tocopherol acetate 〇.〇5 Ethyl p-hydroxybenzoate 0.2 p-Hydroxybenzoic acid Butyl ester 0.1 ACES 1.0 Proper amount (aqueous part) Sodium bisulfite 0.01 Glycerin 5.0 Sodium hyaluronate 0.01 Carboxy vinyl polymer 0.2 Potassium hydroxide 0.2 Refined water remaining -29- 200808366 (26) (Method) 70 °C dissolves the oil phase portion. In part, the oil phase is partially mixed in the water phase, and the machine is cooled by an emulsifier to 3 (TC is manufactured. 7 〇 °C dissolved in the water phase after emulsification] to heat the preparation. Example 6: Anti-wrinkle • Improvement gel

Ingredients (95% ethanol 10.C dipropylene glycol 1 5.C polyethylene oxide (50 moles) oleyl ester 2.1 carboxyvinyl polymer 1.1 caustic soda.: L-sodium succinate. ACES u Methylparaben 〇.: Appropriate amount of ion exchange water remaining in the stomach mash 5 (preparation method) uniformly dissolves the carboxyvinyl polymer. On the other hand, dissolves ACES, polyethylene oxide (50 9 5 % ethanol, Add to the aqueous phase. Then, add other sodium, L-arginine and neutralize to make the viscosity. After ion exchange water, mol) oleyl alcohol after the component, to caustic -30- 200808366 (27) Preparation Example 7: Anti-wrinkle and improving cosmetic ingredients (mass (A phase) Ethanol (9 5 %) 10.0 Polyethylene oxide (20 mol) octyldodecanol 1.0 methylparaben 0.15 ubiquinone ether (pantothenylethylether) 0.1

ACES 0.0 5 (B phase) Potassium hydroxide 0.1 (C phase) Glycerin 5.0 Dipropylene glycol 10.0 Sodium bisulfite 0.03 Carboxy vinyl polymer 0.2 Refining water remaining (preparation method) Dissolve human phase, phase 0, and add human phase It is soluble in the 0 phase. Next, after the phase B is added, the production is carried out. -31 - (28) 200808366 Formulation Example 8: Anti-wrinkle • Improved mask (pack) Ingredient amount (% by mass) (A phase) Dipropylene glycol 5.0 Polyethylene oxide (60 m) hardened castor oil ester 2.0 (B phase) 1.0 5.0 0.2 0.2 0.2

ACES

Olive oil tocopheryl acetate ethyl p-hydroxybenzoate perfume (C phase) sodium bisulfite 0.03 polyvinyl alcohol (saponification degree 90, degree of polymerization 2000) 1 3.0 ethanol 7.0 refined water remaining (preparation method) to uniformly dissolve phase A , phase B, phase C, adding phase B to dissolve in phase A. Next, after the phase C is added thereto, the charging is carried out. -32- (29) 200808366 Compounding amount (% by mass) 2.0 10.0 10.0 40.0 6.0 0.1 0.1 2.0 5.0 10.0 Residual amount of preparation Example 9: Anti-mite and improving ointment component Polyethylene oxide (30 mol) cetyl ether Glycerol monostearate mobile paraffin petrolatum cetyl alcohol p-hydroxybenzoate butyl p-hydroxybenzoate monostearate

ACES Propylene Glycol Ion Exchange Water Fragrance ® (preparation method) Add propylene glycol to ion exchange water, dissolve and heat, and keep at 70 ° C (aqueous phase). Mix 70 ° C to dissolve other components (oil phase). The oil phase is added to the above aqueous phase, uniformly emulsified by a homogenizer, cooled, and then filled. Hereinafter, a cream is produced in the same manner. -33- 200808366 (30) Formulation Example 1 〇 : Anti-wrinkle • Cream for improvement Ingredients (% by mass) Flowing paraffin 8.0 Vaseline 3.0 Dimethyl polyoxane 2.0 Stearyl alcohol 3.0 Dodecanol 2.0

Glycerin 5.0 Dipropylene glycol 4.0 Trehalose 1.0 Tetrapentaerythritol tetrakis 4-ethylhexanoate 4.0 Polyethylene oxide glyceride monoisostearate 2.0 Polyethylene oxide glyceride monostearate 1.0 Lipophilic monostearic acid Glycerate 2.0 Citric acid 0.05 Sodium citrate 0.05 Potassium hydroxide 0.015 Oil soluble licorice extract 0.1 Retinoic acid palmitate (1 million units) 0.25 ACES 1.0 Tocopherol acetate 0.1 Parabens appropriate amount of phenoxyethanol Appropriate amount of dibutylhydroxytoluene amount -34- 200808366 (31) Trisodium ethylenediaminetetraacetate 0.05 4-tert-butyl-4'-methoxydiphenylanthracene methane. 〇1 p-methoxycinnamic acid 2- Ethyl hexyl ester 0.1 /3 -Carotene 0.01 * Polyvinyl alcohol 0.5 ' Hydroxyethyl cellulose 0.5 Carboxyvinyl polymer 0.05 φ Refined water Residual flavor

Formulation Example 1 1 : Anti-wrinkle and improvement component Vaseline dimethyl polyoxyalkyl alcohol ethanol behenyl alcohol Shark liver alcohol glycerol 1,3-butanediol polyethylene glycol 20000 Jojoba oil squalane hydroxyl hard Fatty acid plant sterol ester tetrakis(2-ethylhexanoate) pentaerythritol cream cream (% by mass) 2.0 2.0 5.0 0.5 0.2 7.0 5.0 0.5 3.0 2.0 0.5 1.0 -35- 200808366 (32) Polyethylene oxide hardened castor oil 1.0 Potassium hydroxide 0.1 sodium metabisulfite 0.01 sodium hexametaphosphate 0.05 stearyl glycyrrhizinate 0.1 ubiquinone ether 0.1 arbutin 7.0

传明酸Methyl decylamine hydrochloride 11.0 ACES Potassium salt 1.0 Tocopherol acetate 0.1 Sodium hyaluronate 0.05 Parabens Ethylene triamine tetraacetate 0.05 4-tert-butyl- 4'-A Oxydiphenylmethane methane 0.1 di-p-methoxycinnamic acid mono-2-ethylhexanoic acid glycerin 0.1 yellow iron oxide appropriate amount of Sanxianjiao 0.1 carboxyvinyl polymer 0.2 refined water remaining preparation example 12: anti-wrinkle • improvement Cream ingredients (% by mass) Vaseline 2.0 2.0 Dimethylpolyoxane-36 - 200808366 (33) Ethyl alcohol pentastanol Shark liver alcohol glycerol 1,3-butanediol polyethylene glycol 2000 0 Hobba oil squalane hydroxystearic acid phytosterol ester tetrakilyl 2-ethylhexanoate pentaerythritol ester polyethylene oxide hardened castor oil ester potassium hydroxide sodium metabisulfite sodium hexametaphosphate glycyrrhetinic acid stearyl thiophene ether Fruit glycoside

Benzal acid methyl decylamine hydrochloride ACES tocopheryl acetate sodium hyaluronate p-hydroxybenzoate ethylene diamine tetraacetic acid trisodium 4-tert-butyl _4' methoxy benzophenone methane 5.0 0.5 0.2 7.0 5.0 0.5 3.0 2.0 0.5 1.0 1.0 0.1 0.01 0.05 0.1 0.1 7.0 11.0 1.0 0.1 0.05 Appropriate amount 0.0 5 0.1 -37- 200808366 (34) Di-p-methoxycinnamic acid mono-2-ethylhexanoic acid glyceride 0.1 Yellow oxidation Iron appropriate amount of Sanxianjiao 0.1 carboxyvinyl polymer 0.2 Refined water Remaining manufacturing Examples 1 to 2 Any of the wrinkle-preventing and improving agents are the same as those of Examples 1 and 2 to prevent wrinkles and improve In the effect test, the anti-wrinkle effect is improved. INDUSTRIAL APPLICABILITY The amine-based sulfuric acid compound and the salt of the above formula (1) of the present invention can be used as an anti-wrinkle/improving agent, for example, in the medical product, because it has an excellent anti-wrinkle and improvement effect. Various fields such as cosmetics and pharmaceuticals. [Simplified illustration of the drawings] [Fig. 1] shows the results of the anti-wrinkle/improvement effect test (wrinkle prevention wrinkle mode) by the application of ACES. Fig. 2 is a graph showing changes in skin thickness by coating of ACES. Fig. 3 is a graph showing the results of a wrinkle improvement test (photoaging wrinkle mode) by application of ACES. Fig. 4 is a graph showing the results of a wrinkle improvement test (photoaging wrinkle mode) by application of L-dextrorotatory-α-tocopherol. -38- 200808366 (35) [Fig. 5] A graph showing the results of a wrinkle improvement test (photoaging wrinkle mode) by coating HEPES. Fig. 6 is a graph showing changes in the area ratio of wrinkles by application of ACES.

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Claims (1)

(1) 200808366 X. Patent application scope 1. An anti-wrinkle/improving agent selected from the group consisting of compounds represented by the following formula (1) [Chemical Formula 1] 0 H2N-C-CH2-NH-CH2-CH2 -S 0 0 [Characteristics of containing one or two kinds of aminosulfuric acid compounds and the salt of the salt OH (1) -40-