JP2006273789A - Seramidase inhibitor, and skin external use agents, cosmetics and supplementary medicines containing the inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a seramidase inhibitor used as skin external use agents, cosmetics and supplementary medicines (mild effect medicines controlled by the Ordinance of Ministry of Health, Labor and Welfare) having the effect to improve lowering of skin humidifying barrier function due to stress in particular, and superior in safety and use feeling by improving and maintaining the skin barrier function, relating to the seramidase inhibitor suppressing decomposition of seramide being a lipid between cells by inhibiting seramidase activity in corneous layer and the skin external use agents, cosmetics and supplementary medicines blended with the seramidase inhibitor. <P>SOLUTION: It is characterised in that at least one kind of a vitamin E derivative and N-acyl basic amino acid alkylester or its salts is contained in the seramidase inhibitor. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a ceramidase inhibitor that suppresses the degradation of ceramide, which is an intercellular lipid, by inhibiting ceramidase activity in the stratum corneum, and a skin external preparation, a cosmetic, and a quasi-drug containing the ceramidase inhibitor. About.

  As is well known, ceramide exists in the outermost stratum corneum of the skin and forms a lipid bilayer structure (lamellar structure), thereby transpiration of water from the living body and allergens from the external environment. It is known to play a central role in the skin barrier function that prevents the entry of chemical substances and the like.

  Up to now, to improve the reduced skin barrier function, by applying the synthesized or extracted ceramide itself and activating the ceramide synthesis, the amount of ceramide in the stratum corneum is increased and the skin barrier function is improved. I came. As a technique for applying ceramide, for example, there is a technique using a skin cosmetic as disclosed in Patent Document 1 below, and as a technique for activating ceramide synthesis, for example, ceramide synthesis as described in Patent Documents 2 and 3 below. There are technologies that use accelerators.

JP 2004-168863 A JP 11-322534 A WO02 / 6225 publication

  However, even if ceramide itself is applied using a skin cosmetic as disclosed in Patent Document 1, a lipid bilayer structure cannot be formed in the stratum corneum, so that it is difficult to obtain a sufficient improvement effect. In the case of rough skin due to psychological and social stress, it has been reported that ceramide synthesis in the stratum corneum has already been activated, and therefore, using a ceramide synthesis accelerator as described in Patent Documents 2 and 3 above. Even if ceramide synthesis is activated, the skin barrier function cannot be improved sufficiently.

  The present invention has been made to solve such problems, and has an effect of improving the lowered skin barrier function, and is excellent in safety and usability. It is an object of the present invention to provide a ceramidase inhibitor used as a product.

  As a result of intensive research to solve such problems, the present inventors have made a ceramidase which is an enzyme that degrades ceramide into N-acyl basic amino acid alkyl esters and salts thereof, or vitamin E and derivatives thereof. The present inventors have found that the activity of the skin is specifically suppressed and the skin barrier function is improved, and the present invention has been completed.

  That is, the invention according to claim 1 relating to a ceramidase inhibitor contains at least one of vitamin E, vitamin E derivatives, N-acyl basic amino acid alkyl esters, or salts of N-acyl basic amino acid alkyl esters. It is characterized by.

  The invention according to claim 2 is characterized in that the ceramidase inhibitor according to claim 1 further contains an amphiphilic component. Furthermore, the invention described in claim 3 relating to the external preparation for skin is characterized in that the ceramidase inhibitor described in claim 1 or 2 is blended.

  Furthermore, the invention according to claim 4 relating to cosmetics is characterized in that the ceramidase inhibitor according to claim 1 or 2 is blended. Furthermore, the invention according to claim 5 relating to a quasi-drug is characterized in that the ceramidase inhibitor according to claim 1 or 2 is blended.

  As described above, the present invention comprises a ceramidase inhibitor containing at least one of vitamin E, vitamin E derivatives, N-acyl basic amino acid alkyl esters, or salts of N-acyl basic amino acid alkyl esters. Therefore, by adding such a ceramidase inhibitor to an external preparation for skin, skin cosmetics, quasi drugs, etc., it has a remarkable effect of improving rough skin, and is useful skin cosmetics and pharmaceuticals with excellent usability. It became possible to provide quasi-drugs and external preparations for skin

  As described above, the ceramidase inhibitor of the present invention contains at least one of vitamin E, vitamin E derivatives, N-acyl basic amino acid alkyl esters, or salts of N-acyl basic amino acid alkyl esters. . Here, “contains” means that the ceramidase inhibitor of the present invention is at least one of the above-mentioned vitamin E, vitamin E derivatives, N-acyl basic amino acid alkyl esters, or N-acyl basic amino acid alkyl ester salts. It means that it may consist of or may contain other components.

Examples of vitamin E derivatives include α-tocopherol, β-tocopherol, γ-
Tocopherol, δ-tocopherol, DL-α-tocopherol acetate, DL-α-tocopherol linoleate, DL-α-tocopherol linoleate, DL-α-tocopherol nicotinate, DL-α-tocopherol ascorbate, DL-α phosphate -Tocopherol, DL-α-tocopherol succinate and the like can be used.

  The N-acyl basic amino acid alkyl ester is an alkyl ester in which an acyl group is N-linked to arginine and lysine, which are basic amino acids. In this case, the acyl group preferably has about 8 to 22 carbon atoms.

  Examples of the N-acyl basic amino acid alkyl ester include N-coconut oil fatty acid acyl arginine ethyl ester, N-coconut oil fatty acid acyl arginine propyl ester, N-coconut oil fatty acid acyl arginine butyl ester, N-myristoyl arginine ethyl ester, N -Myristoyl arginine propyl ester, N-myristoyl arginine butyl ester, N-coconut oil fatty acid acyl lysine ethyl ester, N-coconut oil fatty acid acyl lysine propyl ester, N-coconut oil fatty acid acyl lysine butyl ester and the like are exemplified.

  In addition, salts of the N-acyl basic amino acid alkyl ester as described above can also be used. For example, N-acyl basic amino acid alkyl ester glycolate, pyrrolidone carboxylate and the like. More specifically, N-α-coconut oil fatty acid acyl arginine ethyl ester pyrrolidone carboxylate [N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate] (trade name: CAE [Ajinomoto Co., Inc. Company]]) can be preferably used.

  Furthermore, the ceramidase inhibitor of the present invention may further contain an amphiphilic component. Examples of the amphiphilic component include polyoxyethylene surfactants such as polyoxyethylene castor oil, polyglycerol surfactants, derivatives such as phospholipids and lysophospholipids, and polymer emulsifiers.

  As the polymer emulsifier, partially myristoylated chitosan pyrrolidone carboxylate (trade name: PM-chitosan [manufactured by Pierce Co., Ltd.)], alkyl-modified carboxyvinyl polymer (alkyl methacrylate methacrylate copolymer) (trade name: pemlen [ Goodrich, Inc.]), partially myristoylated succinyl chitosan, and the like. The emulsified particle diameter is not particularly limited, but is preferably about 10 to 900 nm. This is because the ceramidase inhibitory action is easily exhibited effectively by making the nanosize to about 10 to 900 nm.

  The amount of N-acyl basic amino acid alkyl ester or salt thereof, vitamin E derivative added to external preparations for skin, cosmetics and quasi drugs is not particularly limited, but is the amount of dry solids, based on the total amount Is preferably 0.0001 to 20% by weight, and more preferably 0.0005 to 5.0% by weight.

  The external preparation for skin, cosmetics, and quasi drugs containing the ceramidase inhibitor of the present invention can be made into dosage forms such as lotions, emulsions, creams, packs, ointments and the like. When the external preparation for skin of the present invention is used as a cosmetic, examples of the cosmetic include cleansing cosmetics, basic cosmetics, hair cosmetics, makeup cosmetics, soaps, and facial cleansers.

In addition, components generally used in external preparations and the like can be widely incorporated in the external preparation for skin, cosmetics, and quasi drugs of the present invention as long as the ceramidase inhibitory action is not inhibited. Such as cetanol,
Higher alcohols such as behenyl alcohol, nonpolar oils such as liquid paraffin and squalane, ester oils such as isopropyl palmitate and isopropyl myristate, vegetable oils such as wheat germ oil and olive oil, trimethylsiloxysilicic acid, methylphenyl poly Silicon compounds such as siloxane and fluorine compounds such as perfluoropolyether can be blended. In addition, pigments, preservatives, surfactants, fragrances, pigments, and the like can be appropriately blended.

  Examples of the present invention will be described below.

[Example 1]
This example is an example of a ceramidase inhibitor, and a ceramidase inhibition test was performed using N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate as a ceramidase inhibitor.

(Extraction of ceramidase)
Ceramidase was extracted according to the following conventional method. The brain extracted from the mouse was homogenized with a buffer solution, and the supernatant obtained by centrifugation was roughly extracted with a buffer solution containing ultrasonic treatment and 0.5% sodium cholate (Merck). This crude extract was subjected to dialysis, ammonium sulfate precipitation, and gel filtration to purify ceramidase and stored frozen at −20 ° C. until test use.

(Preparation of N-coconut oil fatty acyl-L-ethyl alginate-DL-pyrrolidone carboxylate)
N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate was prepared with ultrapure water so as to be 1.0 w / v%. This was diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v%.

(Measurement of ceramidase activity)
The ceramidase activity was measured by allowing the aforementioned ceramidase to act on N-palmitoyl-DL-sphingosine (BIOMOL) as ceramide and measuring the resulting DL-sphingosine level. That is, after N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate was allowed to act on the ceramidase extract, the substrate N-palmitoyl-DL-sphingosine (BIOMOL) suspension was added, The enzyme reaction was carried out at 37 ° C.

  NBD-Cl (4-chloro-7-nitrobenzofurazan: manufactured by Dojindo) ethanol solution was added to this enzyme reaction solution, and DL-sphingosine released in the enzyme reaction solution was subjected to NBD induction reaction at 55 ° C. . After methanol was added to the NBD derivatization reaction solution to stop the reaction, the fluorescence intensity of DL-sphingosine labeled with NBD was measured by HPLC connected with a fluorescence detector at an excitation wavelength of 469 nm and a fluorescence wavelength of 530 nm. To prepare a calibration curve, DL-sphingosine (BIOMOL) sequentially diluted with 40 mg / mL sodium cholate aqueous solution was used, NBD was derived from this, and fluorescence intensity was measured by HPLC.

Such fluorescence intensity was measured using an aqueous solution of N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate at three concentrations of 0.0125, 0.05, and 0.1 w / v%. Was added to the ceramidase extract, and when N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate was not added, and the ratio of the fluorescence intensity measured in each case was determined. Asked. The results are shown in Table 1.

  In Table 1, for the ceramidase activity in each test, the ceramidase activity when N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate was not added was taken as 100, and the ratio of the fluorescence intensity Based on the figure, the value of ceramidase activity is shown. As shown in Table 1, ceramidase activity decreased depending on the addition concentration of N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylic acid. From this test result, N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate was found to have an effect of suppressing ceramidase activity.

[Example 2]
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using a vitamin E derivative as a ceramidase inhibitor. In this example, DL-α-tocopherol acetate was used as the vitamin E derivative.

(Extraction of ceramidase)
The extraction of ceramidase was performed in the same manner as in Example 1. This vitamin E derivative was dissolved in ethanol to prepare 1.0 w / v%, and further diluted with ultrapure water to give 0.005, 0.01, and 0.02 w / v%.

(Measurement of ceramidase activity)
The ceramidase activity was measured in the same manner as in Example 1. The test results of Example 2 are shown in Table 2.

表２に示すように、ビタミンＥ誘導体を作用させることによって、セラミダーゼ活性はビタミンＥ誘導体の添加濃度に依存して減少した。この試験結果から、ビタミンＥ誘導体に、セラミダーゼ活性を抑制する作用が認められた。

As shown in Table 2, the action of the vitamin E derivative decreased the ceramidase activity depending on the added concentration of the vitamin E derivative. From this test result, the vitamin E derivative was confirmed to have an effect of suppressing ceramidase activity.

[Examples 3 and 4 and Comparative Examples 1 to 3]
In this example, the effect of N-acyl basic amino acid alkyl ester salts, which are ceramidase inhibitors shown in Examples 1 and 2 above, on the skin barrier function of vitamin E derivatives was tested.

(Preparation of salts of N-acyl basic amino acid alkyl ester)
Example 3 was a solution in which N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate was dissolved in a 10% ethanol solution to 5%.

(Preparation of vitamin E derivatives)
A solution obtained by dissolving DL-α-tocopherol acetate in a 10% ethanol solution to 5% was defined as Example 4.

(Test method)
BALB / c mice male with shaved back, 7 weeks old, were reared in 40 animals / 1 cage for 10 days and overstressed. The ceramidase inhibitors of Examples 3 and 4 were applied to the back of mice shaved daily during overload stress loading. After releasing from overstress stress, measure the transdermal water transpiration of the back skin of the mouse, the amount of ceramide per stratum corneum area, and the amount of ceramide per total lipid in the stratum corneum. Then, the state of the tissue was observed.

  In addition, the measurement and observation were similarly performed by using as a comparative example 1 the mouse | mouth of the non-stress load which did not load the overstress stress reared by 5 animals / cage during the test period. In addition, the case where only the stress is applied to the mouse but the ceramidase is not applied is referred to as Comparative Example 2, and the case where only the 10% ethanol solution which is the base of Examples 3 and 4 is applied to the mouse under the overcrowded stress is shown as Comparative Example 3. It was.

(Measurement of barrier function)
The transdermal moisture transpiration of the mouse dorsal skin of Examples 3 and 4 and Comparative Examples 1 to 3 was measured with Tevameter TM210 (Integral), and the difference between the measured values of Comparative Example 1 and Comparative Example 2 was converted to 100. The differences between Examples 3 and 4 and Comparative Example 3 and Comparative Example 1 are shown as TEWL recovery rates. Table 3 shows the measurement results of the recovery rate of the barrier function.

  As is clear from Table 3, the skin of the mouse to which the ceramidase inhibitor of Examples 3 and 4 was applied was restored to the barrier function compared to the skin of the mouse to which the base of Comparative Example 3 was applied. I understood. This indicates that the skin barrier function was improved by applying the ceramidase inhibitor of Examples 3 and 4 to the skin.

(Measurement of the amount of ceramide in the stratum corneum)
The stratum corneum was collected from the back skin of Examples 3 and 4 and Comparative Example 3 with cyanoacrylate resin, and lipid was extracted with a hexane: ethanol (95: 5) solution. The extracted lipids were separated by thin layer chromatography, and after coloring, the density of each spot that appeared was quantified, and expressed as the amount of ceramide per area of the collected stratum corneum and the ratio of the amount of ceramide to the total amount of lipid extracted. Table 4 shows the amount of ceramide per stratum corneum area, and Table 5 shows the ratio of the amount of ceramide to the total amount of lipid in the stratum corneum.

  As is clear from Table 4, the mouse skin in which the ceramidase inhibitor of Examples 3 and 4 was applied to the skin was more per stratum corneum in all Examples than the case of applying the base of Comparative Example 3. It was found that the amount of ceramide increased. This indicates that application of Examples 3 and 4 to the skin suppresses a decrease in the amount of ceramide in the stratum corneum due to stress.

  Furthermore, also in Table 5, the mouse skin which apply | coated the ceramidase inhibitor of Example 3, 4 compared with the case where the base which is the comparative example 3 was apply | coated with respect to the total lipid amount in a stratum corneum in all Examples. It was found that the ratio of the amount of ceramide tends to increase. This suggests that the decrease in the amount of ceramide in the stratum corneum due to stress is not due to the decrease in intercellular lipids, but the decrease in ceramide itself.

(Observation of tissue state)
The dorsal skin excised from the mice of Examples 3 and 4 and Comparative Examples 1 to 3 was fixed in 20% formalin neutral buffer, and then a histopathological specimen stained with hematoxylin and eosin was prepared and observed under a microscope. As for the observation of the tissue, when the degree of epithelial layer thickening, basal cell spindle formation, stratum corneum stratification, and keratinization failure are markedly observed, ++, and when 80% of the total are observed, + The case where a part is observed is represented by ±, and the case where it is not observed is represented by −. Table 6 shows the observation results of the tissue state.

From the observation of the tissue state shown in Table 6, it was found that keratinization abnormalities in the epidermis layer were suppressed in the skin applied with Examples 3 and 4 as compared with Comparative Examples 2 and 3. This indicates that application of the ceramidase inhibitor of Examples 3 and 4 improves keratinization abnormality caused by stress and maintains a more normal state.

  From the measurement results of the barrier function, the measurement result of the ceramide amount per stratum corneum, the measurement result of the ratio of the ceramide amount to the total lipid amount in the stratum corneum, the observation result of the tissue state, etc., the N-acyl basicity N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate, which is one of the salts of amino acid alkyl esters, and DL-α-tocopherol acetate, which is one of the vitamin E derivatives, contain ceramide due to stress. It was found that there is an effect of suppressing the decrease and maintaining the skin barrier function in a more normal state.

(Skin roughness improvement test)
This example is a formulation example in the case of blending N-acyl basic amino acid alkyl ester salts, vitamin E derivatives, which are the ceramidase inhibitors, into creams as an example of cosmetics. went.

  The cream was prepared as follows. Squalene, ceralkyl alcohol, cetyl isooctanoate, and microcrystalline wax are heated and dissolved, and then added with clay mineral and POE glycerol triisostearate, adjusted to 70 ° C, and uniformly dispersed and dissolved to obtain an oily gel. It was. N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate and vitamin E derivative are dissolved in purified water of a predetermined concentration, adjusted to 70 ° C., and then sufficiently stirred into the oily gel. Slowly added. After uniformly mixing with a homomixer, the mixture was deaerated and filtered, and then cooled to 30 ° C. to obtain a cream. The composition and blending ratio of the creams of Examples 5 and 6 obtained are as follows.

Example 5
Composition ratio (wt%)
Squalene 20%
Serakil alcohol 2.0%
Cetyl isooctanoate 8.5%
Microcrystalline wax 1%
Clay mineral 1.3%
POE glycerol triisostearate 0.2%
Diglycerin 2.0%
N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate 0.5%
Chitosan pyrrolidone carboxylate 0.2%
Residual amount of water

Example 6
Composition ratio (wt%)
Squalene 20%
Serakil alcohol 2.0%
Cetyl isooctanoate 8.5%
Microcrystalline wax 1%
Clay mineral 1.3%
POE glycerol triisostearate 0.2%
Diglycerin 2.0%
Vitamin E derivative (DL-α-tocopherol acetate) 0.5%
Chitosan pyrrolidone carboxylate 0.2%
Water remaining

Using the creams of Examples 5 and 6 prepared in this manner, a rough skin practical test was conducted. The test method is as follows. For 30 female subjects (25 to 45 years old) who complain of rough skin and dry skin, a sample and a cream base were applied to the affected area half a day twice a day for 1 month.
Evaluation was shown by the number of people who answered that the degree of improvement of rough skin and dry skin was better in the sample application site than in the cream base application site.

  On the other hand, the same test as Comparative Example 4 was performed on a cream containing no ceramidase inhibitor such as N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate or vitamin E derivative. The test results are shown in Table 7.

  As is apparent from Table 7, in Examples 5 and 6, the number of respondents who felt moist was about three times or more than that of Comparative Example 4.

[Examples 7 and 8]
This example is a formulation example in the case of blending a salt of N-acyl basic amino acid alkyl ester, which is the ceramidase inhibitor, and a vitamin E derivative into a lotion as an example of a cosmetic, and the rough skin of the lotion A practical test was conducted.

  The lotion was prepared as follows. Preparation of a lotion base containing surfactant POE (20) oleyl alcohol ether, thickener methylcellulose and quince seed, ethanol, N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylic acid at a predetermined concentration Acid salts or vitamin E derivatives were added. The compositions and blending ratios of the obtained lotions of Examples 7 and 8 are as follows.

Example 7
Composition ratio (wt%)
POE (20) oleyl alcohol ether squalene 0.5%
Partially myristoylated chitosan pyrrolidone carboxylate 0.1%
Methylcellulose 0.2%
Quince Seed 0.1%
Ethanol 10%
Pentadiol 2.0%
N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate 0.5%
Water remaining

Example 8
Composition ratio (wt%)
POE (20) oleyl alcohol ether squalene 0.5%
POE hardened castor oil 0.2%
Partially myristoylated carboxymethyl chitosan 0.2%
Methylcellulose 0.2%
Quince Seed 0.1%
Ethanol 10%
Pentadiol 2.0%
Vitamin E derivative (DL-α-tocopherol acetate) 0.5%
Vitamin E derivative (DL-α-tocopherol nicotinate) 0.2%
Water remaining

  Using the lotions of Examples 7 and 8 prepared in this way, a rough skin practical test was conducted. The test method is as follows. For 30 female subjects (25 to 45 years old) who complain of rough skin and dry skin, a sample and a lotion base were applied to the affected area half a day twice a day for 1 month. Evaluation was shown by the number of people who answered that the degree of improvement of rough skin and dry skin was better in the sample application part than in the lotion base application part. On the other hand, the same test as Comparative Example 5 was performed on a lotion containing no ceramidase inhibitor. The test results are shown in Table 8.

  As is apparent from Table 8, in Examples 7 and 8, the number of respondents who felt moist was about 1.5 to 2 times that of Comparative Example 5.

Example 9
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was conducted using α-tocopherol as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. α-tocopherol was dissolved in ethanol to prepare 1.0 w / v%, and further diluted with ultrapure water to obtain 0.005, 0.01, and 0.02 w / v%. The ceramidase activity was measured in the same manner as in Example 1.

Example 10
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using DL-α-tocopherol linoleate as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. DL-α-tocopherol linoleate is dissolved in ethanol to prepare 1.0 w / v%, and further diluted with ultrapure water to obtain 0.005, 0.01, and 0.02 w / v. %. The ceramidase activity was measured in the same manner as in Example 1.

Example 11
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using DL-α-tocopherol linoleate as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. DL-α-tocopherol linolenic acid was dissolved in ethanol to prepare 1.0 w / v%, which was further diluted with ultrapure water to give 0.005, 0.01, and 0.02 w / v. %. The ceramidase activity was measured in the same manner as in Example 1.

Example 12
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using DL-α-tocopherol nicotinate as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. Nicotinic acid DL-α-tocopherol is dissolved in ethanol to prepare 1.0 w / v%, and further diluted with ultrapure water to give 0.005, 0.01, and 0.02 w / v. %. The ceramidase activity was measured in the same manner as in Example 1.

Example 13
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using DL-α-tocopherol ascorbate as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. Ascorbic acid DL-α-tocopherol was dissolved in ethanol to prepare 1.0 w / v%, which was further diluted with ultrapure water to give 0.005, 0.01, and 0.02 w / v. %. The ceramidase activity was measured in the same manner as in Example 1.

Example 14
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using DL-α-tocopherol succinate as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. DL-α-tocopherol succinate was dissolved in ethanol to prepare 1.0 w / v%, and further diluted with ultrapure water to give 0.005, 0.01, and 0.02 w / v. %. The ceramidase activity was measured in the same manner as in Example 1.

  Table 9 shows the test results of Examples 9 to 14.

  As is clear from Table 9, the ceramidase activities of Examples 9 to 14 decreased depending on the concentration of each ceramidase inhibitor added. From this test result, the ceramidase inhibitor of each Example 9-14 was confirmed to have an action of suppressing ceramidase activity.

Example 15
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using N-coconut oil fatty acyl arginine propyl ester as a ceramidase inhibitor.
The extraction of ceramidase was performed in the same manner as in Example 1. N-coconut oil fatty acid acyl arginine propyl ester was prepared with ultrapure water so as to be 1.0 w / v%, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v. v%. The ceramidase activity was measured in the same manner as in Example 1.

Example 16
This example is another example of a ceramidase inhibitor, and a ceramidase inhibition test was performed using N-coconut oil fatty acyl arginine butyl ester as a ceramidase inhibitor.
The extraction of ceramidase was performed in the same manner as in Example 1. N-coconut oil fatty acyl arginine butyl ester was prepared with ultrapure water so as to be 1.0 w / v%, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v. v%. The ceramidase activity was measured in the same manner as in Example 1.

Example 17
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using N-myristoylarginine ethyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-myristoyl arginine ethyl ester was prepared at 1.0 w / v% and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v%. The ceramidase activity was measured in the same manner as in Example 1.

Example 18
This example is another example of a ceramidase inhibitor, and a ceramidase inhibition test was performed using N-myristoylarginine butyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-myristoyl arginine butyl ester was prepared with ultrapure water to 1.0% w / v, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v%. did. The ceramidase activity was measured in the same manner as in Example 1.

Example 19
This example is another example of a ceramidase inhibitor, and a ceramidase inhibition test was performed using N-coconut oil fatty acid acyl lysine ethyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-coconut oil fatty acid acyl lysine ethyl ester was prepared to 1.0 w / v% with ultrapure water, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v. v%. The ceramidase activity was measured in the same manner as in Example 1.

Example 20
This example is another example of a ceramidase inhibitor, and a ceramidase inhibition test was performed using N-coconut oil fatty acid lysine propyl ester as a ceramidase inhibitor.
The extraction of ceramidase was performed in the same manner as in Example 1. N-coconut oil fatty acid acyl lysine propyl ester was prepared to be 1.0 w / v% with ultrapure water, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v. v%. The ceramidase activity was measured in the same manner as in Example 1.

Example 21
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was conducted using N-coconut oil fatty acid acyl lysine butyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-coconut oil fatty acid acyl lysine butyl ester was prepared with ultrapure water so as to be 1.0 w / v%, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v. v%. The ceramidase activity was measured in the same manner as in Example 1.

[Example 22]
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using N-myristoyllysine ethyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-myristoyl lysine ethyl ester was prepared to 1.0 w / v% with ultrapure water, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v%. did. The ceramidase activity was measured in the same manner as in Example 1.

Example 23
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using N-myristoyl lysine propyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-myristoyl lysine propyl ester was prepared with ultrapure water to be 1.0 w / v%, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v%. did. The ceramidase activity was measured in the same manner as in Example 1.

Example 24
This example is another example of a ceramidase inhibitor. A ceramidase inhibition test was performed using N-myristoyllysine butyl ester as a ceramidase inhibitor. The extraction of ceramidase was performed in the same manner as in Example 1. N-myristoyl lysine butyl ester was prepared with ultrapure water so as to be 1.0 w / v%, and further diluted with ultrapure water to 0.0125, 0.05, and 0.1 w / v%. did. The ceramidase activity was measured in the same manner as in Example 1.

  The test results of Examples 15 to 24 are shown in Table 10.

  As is clear from Table 10, the ceramidase activities of Examples 15 to 24 decreased depending on the concentration of each ceramidase inhibitor added. From this test result, the ceramidase inhibitor of each Example 15-24 was confirmed to have an action of suppressing ceramidase activity.

(Prescription Example 1)
This formulation example is a formulation example of a translucent lotion for improving the skin barrier. Its composition is as follows.

Composition ratio (wt%)
(1) Butylene glycol 1.0%
(2) N-coconut oil fatty acid acyl arginine ethyl ester pyrrolidone carboxylate 0.2%
(3) N-coconut oil fatty acid acyl lysine ethyl ester pyrrolidone carboxylate 0.1%
(4) Vitamin E derivative dispersion (average particle size: 200 nm) 7.5%
Composition: Tocopherol acetate 0.08%
Tocopherol succinate 0.02%
Pentadiol 2.0%
POE hardened castor oil 0.1%
Lysophospholipid 0.1%
Partial myristoylated chitosan
Pyrrolidone carboxylate 0.2%
Purified water 5.0%
(5) Glycerin 10.0%
(6) Glycolic acid 0.1%
(7) Chitosan glycolate 0.1%
(8) Glutathione 0.1%
(9) Ethanol 5.0%
(10) Purified water remaining

  The case of preparing the lotion of the present formulation example will be described. First, the component (4) above is subjected to high pressure treatment (2000 psi) with a high pressure fluidizer device to obtain a vitamin E derivative dispersion (average particle size: 200 nm). It was. Next, the dispersion liquid of (4) is added to the heat-treated components (1) to (3) and uniformly dissolved in the components (5) to (8) to obtain a translucent essence lotion. It was.

(Prescription example 2)
This formulation example is a formulation example of an external cream for improving the skin barrier. Its composition is as follows.

Composition ratio (wt%)
(1) Seraquil alcohol 3.5%
(2) Squalane 4.0%
(3) Sorbitan monostearate 3.0%
(4) Glycosylceramide 0.2%
(5) Vitamin E derivative dispersion (average particle size: 300 nm) 10.0%
Composition: 0.2% tocopherol acetate
γ-tocopherol 0.2%
Butanediol 2.0%
Polyoxyethylene castor oil 0.1%
Hydroxylated phospholipid 0.1%
Partially myristoylated carboxy
Methyl chitosan 0.2%
Purified water 7.2%
(6) N-coconut oil fatty acid acyl lysine ethyl ester pyrrolidone carboxylate 0.05%
(7) N-myristoylarginine ethyl ester pyrrolidone carboxylate 0.02%
(8) Citrulline 1.5%
(9) Rosemary extract 1.0%
(10) Glutathione 0.5%
(11) Methylparaben 0.2%
(12) Remaining amount of purified water

  The case of preparing the cream of this formulation example will be described. First, the vitamin E derivative dispersion (average particle size: 300 nm) is obtained by high-pressure treatment (2500 psi) of the composition component (5) with a high-pressure fluidizer. It was. Next, the dispersion liquid of (5) is added to the heat-treated water phase composition of (6) to (12), and the oil phase of (1) to (4) which is heated and dissolved is added. A creamy preparation was obtained by emulsification at 8000 rpm for 1 min.

[Other Examples]
In addition, although the said Example demonstrated the case where only 1 type of N-coconut oil fatty acid acyl-L-ethyl alginate-DL-pyrrolidone carboxylate and a vitamin E derivative was used, 2 or more types were mix | blended. It is also possible to use it.

Claims (5)

  A ceramidase inhibitor comprising at least one of vitamin E, a vitamin E derivative, an N-acyl basic amino acid alkyl ester, or a salt of an N-acyl basic amino acid alkyl ester.   The ceramidase inhibitor according to claim 1, further comprising an amphiphilic component.   An external preparation for skin, comprising the ceramidase inhibitor according to claim 1 or 2.   A cosmetic comprising the ceramidase inhibitor according to claim 1 or 2.   A quasi drug containing the ceramidase inhibitor according to claim 1 or 2.