KR20010044801A - N-acetylphytosphingosine retin amide derivatives and preparation process thereof - Google Patents

Abstract

PURPOSE: Provided is an acetylated N-acetyl phytosphingosine retinamide derivative, which can be used as an inhibitor against skin aging, wrinkles, skin-roughness, skin-spot and as a functional material of medicine or cosmetics for atopy, pimples, psoriasis, skin-dryness, antimicrobe, and antiinflammation. CONSTITUTION: The acetylated N-acetyl phytosphingosine retinamide derivative (formula 1) is produced by reacting N-acetyl phytosphingosine and a retinoic acid derivative in the presence of a condensing agent selected from 1,3-dicyclohexyl carbodiimide(DCC) or N,N-carbonyl diimidazol(CDI) and an organic amine catalyst to obtain N-acetyl phytosphingosine retinamide derivative, followed by reacting the N-acetyl phytosphingosine retinamide derivative with acetic anhydride.

Description

Retinamide derivatives and preparation process of N-acetylphytosphingosine {N-acetylphytosphingosine retin amide derivatives and preparation process

The present invention relates to retinamide derivatives of N-acetylphytosphingosine. More particularly, the present invention relates to acetylated N-acetylphytosphingosine retinamide derivatives useful as skin aging inhibitors and methods for preparing the same.

The acetylated N-acetylphytosphingosine retinamide derivatives of the general formula 1 according to the present invention are not only non-irritating but also exhibit enhanced stability, and thus can be usefully used as functional raw materials for anti-aging medicines and cosmetics.

Wrinkles, roughness and blemishes from photoaging have been reported to be improved by the application of creams containing retinoids, but these substances are sensitive to temperature, oxidation and light and are unstable and irritating to the skin and during the incubation period. May cause side effects such as dryness, cuts, and flaking. Due to these side effects, there are many difficulties in using retinoid as a main ingredient of medicines and cosmetics. Therefore, there is an urgent need for the development of non-irritating new functional raw materials while maintaining the activity of retinoids such as suppressing photoaging.

In addition, there is a need for a new skin aging inhibitor that has improved stability and maximizes cell regeneration in vivo. It has already been reported that retinyl acetate and retinyl palmitate obtained by reacting vitamin A alcohol (retinol) with acetic acid or palmitic acid have improved instability and irritation compared to retinol (Kretz. A., SOFW (1993)). 119.pp 21-24).

However, the role of retinoid-based skin improvement is known to be best regulated by retinoic acid, and the activity of retinol, retinal and other vitamin A derivatives is dependent on the conversion to retinoic acid. Therefore, two steps of enzymatic conversion (hydrolysis of ester bonds and oxidation of retinol) are required to see the activity of retinoic acid in such retinol derivatives.

In particular, since the above-mentioned compounds are fat-soluble, the skin absorption ability is inherently weak, so it is difficult to greatly expect the activity of retinoic acid such as enhancing collagen synthesis, preventing elastin degradation, and regeneration of epidermal cells. Polyethoxylated retinamide derivatives have been reported using polyethylene glycol which is excellent in hydrophilic activity and hydrophilic in order to increase skin absorption from the skin (see Korean Patent Publication No. 99-77401).

However, these derivatives, like acetate and palmitate, do not have the added benefit of having polyethylene glycol act on the skin independently.

On the other hand, ceramide is used worldwide for skin care cosmetics due to its excellent moisturizing function and regeneration of damaged skin protective film, and its use has been extended to hair products.

Phytosphingosine and N-acetylphytosphingosine have been used as functional raw materials such as moisturizers and anti-aging agents in addition to ceramides. The use of phytosphingosine and N-acetyl phytosphingosine is increasing rapidly due to its excellent skin regeneration, antibacterial and anti-inflammatory effects.

The technical problem to be achieved by the present invention is a ceramide, which occupies 20 to 40% of the skin lipids, the basic structure of sphingosine base, sphingosine base, sphingosine base, sphingosine base, and fatty acid connected to amino group of sphingolipid base. With this in mind, a new acetylated N-acetylpytosphingosine retinamide derivative of general formula (1) in which an amide condensation reaction of N-acetylphytosphingosine with a retinoic acid derivative is developed.

The derivative developed by the present invention is an amide condensation reaction of N-acetylphytosphingosine with retinoic acid (all-trans retinoic acid) or retinoic acid isomer (e.g., 9-cis-retinoic acid or 13-cis-retinoic acid). Contains substances. It is also possible to develop derivatives composed of retinol. That is, one carboxyl portion of the dicarboxylic acid and the retinol may be ester-bonded, and the other carboxyl portion may form an amide bond with N-acetylphytosphingosine.

The substance of the present invention produces not only retinoic acid but also N-acetylphytosphingosine by enzymatic hydrolysis. Therefore, the N-acetylphytosphingosine is expected to additionally expect its bioactive characteristics as a regulatory substance for epidermal cell growth and differentiation.

In addition, retinoic acid has a lot of difficulties due to solubility problems in using a sufficient amount to achieve the optimum effect. The retinamide of N-acetylphytosphingosine is increased by 2-3 times solubility through the acetylation reaction to solve the solubility problem to some extent and is expected to be used as a functional raw material in anti-aging medicines and cosmetics.

Figure 1 shows the result of comparing the collagen synthesis effect between the compound of the present invention and retinoic acid and the control.

Figure 2 shows the results of the antimicrobial test between the acetylated N-acetylphytosphingosine retinamide and erythromycin and the control compound of the present invention.

Therefore, an object of the present invention is to use the N-acetyl phytosphingosine derivative of the general formula 2 as a starting material;

(Formula 2)

In the formula, X means hydrogen or leaving group.

Reacting with a retinoic acid derivative of the following general formula (3) in the presence of a condensing agent selected from 1,3-dicyclohexylcarbodiimide (DCC) or N, N-carbonyldiimidazole (CDI) and an organic amine catalyst;

(Formula 3)

HO-Q

In the above formula, Q means a retinoic acid moiety of the following structural formulas (Q-1, Q-2, Q-3).

(Q-1)

(Q-2)

(Q-3)

To prepare an N-acetylphytosphingosine retinamide derivative of the following general formula (4);

(Formula 4)

This is reacted with acetic anhydride again to provide a method for preparing the acetylated N-acetylphytosphingosine retinamide derivative of the general formula (1).

(Formula 1)

The present invention also provides an acetylated N-acetylphytosphingosine retinamide derivative of the general formula (1).

On the other hand, the present invention is the acetylated N-acetyl phytosphingosine retinamide derivatives, wrinkles, skin roughness, skin moisturizing, anti-aging and photoaging, such as atopic dermatitis, acne, psoriasis, dry skin, antibacterial, anti-inflammatory for the purpose of medicine or cosmetics Provides a method to use as a raw material.

On the other hand, cosmetics, such as cosmetic creams, essences, packs, skins containing 0.01 to 10% by weight of the acetylated N-acetylphytosphingosine retinamide derivatives are provided.

The compound of formula 1 of the present invention has a structure in which the acyl group of ceramide corresponds to the retinoid as a ceramide analogue. The compound is more chemically stable and safer than conventional retinoic acid, and can be penetrated into the stratum corneum more quickly and effectively due to its improved lipid-friendly properties. When it reaches the stratum corneum, the compound releases retinoic acid by various enzymes in the dermis. Eventually, the compound according to the present invention can effectively penetrate retinoic acid into the skin.

Hereinafter, a method for preparing the acetylated N-acetylphytosphingosine retinamide derivative of the present invention will be described.

As the organic amine serving as a catalyst, triethylamine is preferably used, and the amount thereof is preferably 1.0 to 1.5 molar amount based on the compound of the general formula (2). As a condensing agent, 1,3-dicyclohexylcarbodiimide (DCC) or N, N-carbonyldiimidazole (CDI) is used, and the reaction is usually carried out in a state in which light and water are blocked and is performed at room temperature.

However, the catalysts and condensing agents which can be used in the process according to the invention are not limited to those listed above, and all conventionally known in the art can be used within the scope that does not affect the reaction. In addition, the compound of the general formula 1 prepared by the above-described method can be purified by a conventional separation and purification method, for example, recrystallization or extraction.

The compound of Formula 1 may be used for the treatment of inflammation, wrinkles, freckles and the like caused by skin diseases such as skin cancer or acne. In particular, the compound of Formula 1 is characterized in that compared to retinoic acid has no skin irritation according to the treatment and has the effect of antibacterial, anti-inflammatory properties of N-acetylphytosphingosine.

Experiments to confirm the efficacy of the compound of formula 1 is carried out as follows.

First, the skin irritation experiment is performed at a concentration in the range of 0.1 to 1% through a patch experiment using a guinea pig (Guinea Pig).

As a result of these experiments, the compound of formula 1 according to the present invention is compared to retinoic acid, which is an anti-aging pharmaceutical and cosmetic raw material, which has been commercially used in the past, and the compound of formula 1 has an antimicrobial and anti-inflammatory property in addition to its hypoallergenicity and improved stability. Since it has the physiologically active function of N-acetylphytosphingosine, it can be usefully used as a multifunctional additive in medicine and cosmetics.

Hereinafter, the present invention will be described in more detail based on the following Examples and Experimental Examples. However, these Examples and Experimental Examples are only for better understanding of the present invention, and the scope of the present invention in any sense is not limited to these Examples.

Example 1 Preparation of N-acetylphytosphingosine Retinamide Derivative of General Formula 4

24.9 g (0.0693 mol) of N-acetylphytosphingosine was dissolved in 200 ml of anhydrous chloroform, and then 25 g (0.0832 mol) of retinoic acid and 8.42 g (0.0832 mol) of triethylamine were added thereto, followed by 1,3-dicyclohexylka. The solution to which the body imide was added was stirred at room temperature for 1 hour while blocking light and moisture. The reaction solution was extracted with water and washed. The separated organic layer was concentrated by distillation under reduced pressure and recrystallized in 200 ml of ethanol at 4 ° C. for 2 hours to remove remaining retinoic acid and unreacted N-acetylphytosphingosine. The precipitate was filtered off and dried to give 33.4 g (yield 75%) of the pale yellow title compound.

Example 2 Preparation of Acetylated N-acetylphytosphingosine Retinamide Derivative of General Formula 1

25 g (0.0389 mol) of N-acetylphytosphingosine retinamide was dissolved in 200 ml of anhydrous chloroform, and 7.88 g (0.0778 mol) of triethylamine and 11.94 g (0.1169 mol) of acetic anhydride were added to block light and moisture. The mixture was stirred at room temperature for 4 hours. The reaction solution was washed with water, the separated organic layer was concentrated by distillation under reduced pressure, and then recrystallized from 200 ml of ethanol at 4 ° C. for 2 hours. The precipitate was filtered off and dried to give 24.0 g (yield 90.1%) of the title compound as pale yellow.

Example 3 Acetylated N-acetylphytosphingosine Retinamide Skin Irritation Test

Experiments were performed using 6-week-old BALB / C mice to test the degree of skin primary irritation. In order to apply the test substance to more than 10% of the total surface area of the test 24 hours before the start of the test, the hairs of 10 test animals and the like were cut carefully with 6 × 4 cm 2 area to prevent damage to the skin.

After 24 hours of epilation, apply a solution of 0.3% Napol, Retinoic Acid, and Retinyl Palmitate sample solution using DMSO as a soluble solvent, and then cover the entire application site with a thin, uniform porous gauze for 24 hours. The contact between the test substance and the skin was maintained. After removing the patch, on the day of administration, the general condition was observed every hour up to 12 hours, and carefully observed once daily from the next day to 14 days to determine the degree of stimulation. The results are shown in Table 1 below.

sample Stimulus index Stimulus DMSO 2.0 Severe Stimulation DMSO + Retinoic Acid 1.9 Severe Stimulation DMSO + Retinyl Palmitate 1.5 Weak irritation DMSO + Compound of Formula 1 0.9 Fine stimulation

As can be seen from the results in Table 1, it can be seen that there is a significant difference in the degree of stimulation between the retinoic acid and the compound of the general formula 1 according to the present invention.

Example 4 Human Patch Experiment

In order to determine the skin irritation of the sample (Napol) prepared in the above example, a certain amount (approximately 40 µg) was applied to the upper and lower portions of the healthy adult men and women and the pin chamber was removed for 2 hours, and then a plurality of The expert performed a visual judgment.

sample Examinee Judgment result Irritation degree ^** (%) ++ + +- - Napol 0.3% 30 0 0 2 28 6.67 Napol 0.5% 30 0 0 3 27 10.00 Purified water 30 0 0 One 29 3.33 Retinoic acid 0.3% 30 One 2 5 22 35.29 Retinyl Palmitate 0.3% 30 0 One 4 25 19.35

* Criteria-: No erythema or unusual phenomenon

+-: Slightly reddish

+: Significantly redder than the surroundings

++: Severe reddening and swelling around

** Stimulus (%) = {{{LEFT {(+-) number TIMES 1 RIGHT}}} + {{LEFT {(+) number TIMES 2 RIGHT}}} + {{LEFT {(++) number TIMES 3 RIGHT}}}} OVER {{{LEFT {(+-) number TIMES 1 RIGHT}}} + {{LEFT {(+) number TIMES 2 RIGHT}}} + {{LEFT {(++) number TIMES 3 RIGHT}}} + (-)} TIMES 100

As can be seen from the results in Table 2, the skin primary irritation was low.

Example 5 Collagen Synthesis Effect Experiment

In order to confirm the utility of the compound of Formula 1 as a skin aging inhibitor, its collagen synthesis effect was compared using retinoic acid as a control. The same number of human fibroblasts (Human Fibroblast) were incubated for 24 hours on a plate having 24 wells, and samples were applied at 10 ⁻⁴ to 10 ⁻⁷ W% / ml. 2 hours later, tritium-induced proline (L-2,3-3H proline) was incubated for 24 hours, and the amount of collagen newly synthesized by fibroblasts was cultured using culture medium. The amount of csp (collagenous sensitive protein) was calculated by measuring on the basis of (counting per minute). In addition, the results of comparing the collagen synthesis effect using the calculated csp amount is shown in Figure 1 below (Reference: 1. David FW and Wilson Harvey, Analytical Biochemistry, 1979; 96, 220-224. 2. Atsushi Hatamochi, Masashi Ono, Hiroaki Ueki and Masayoshi Mamba., J. Invest Dermatol., 1991, 96, 473-477). However, in FIG. 1, the control group indicates that the skin aging inhibitor was not added, and serum having activity for collagen synthesis was added.

As shown in Figure 1, retinoic acid showed an increase of up to 20.7% at 10 ^-6 W% / ㎖. And the compound of Formula 1 showed up to 27.8% enhancement effect at 10 ^-7 W% / ㎖.

Example 6 Stability Test

After 6 months at room temperature and 40 ℃ storage conditions, the degree of titer storage was compared with retinoic acid.

Storage condition Napol Retinoic acid Room temperature 98% 73% 40 ℃ 95% 49%

As can be seen from the results of Table 3, it can be seen that Napol shows nearly twice the stability of retinoic acid.

Example 7 Acetylated N-acetylphytosphingosine Retinamide Antibacterial Activity Test

In order to measure the antimicrobial activity of acetylated N-acetylphytosphingosine retinamide against skin harmful bacteria, the antimicrobial activity was tested against Propionibacterium acnes, the cause of acne. The medium used to test antimicrobial activity against Propioni bacterium acnes was 25 g of Brain Heart Infusion, 5 g of yeast extract, 4 g of Casitone, 1 g of L-cysteine HCl, 5 g of glucose, 1 g of soluble starch in 1 liter of distilled water. , 15 g of Monopotassium Phosphate, 1 g of ammonium sulfate, 0.2 g of magnesium sulfate, and 0.02 g of calcium chloride were dissolved evenly and used after autoclaving. After the anaerobic condition was formed using BBL GasPak Anaerobic System, about 3 ~ 5 days at 37 ℃ Incubated. After the incubation, the number of bacteria was measured to check the antimicrobial activity. Samples of acetylated N-acetylphytosphingosine retinamide used in the present invention were used by dissolving ethanol, and in order to determine the antimicrobial effect, at concentrations of 1 μg / ml, 10 μg / ml, 100 μg / ml and 1,000 μg / ml, Used. Each of the microorganisms were cultured to determine the dilution factor to form a ^3-10 10 ⁴ colonies were cultured and plated on each of the culture medium per plate medium was diluted 10-fold in sequence. Each microorganism was incubated and then diluted with the dilution factor determined in the above experiment (0.85% NaCl was used as the dilution solution). After diluting the sample prepared by the method described above in the solvent used for sample preparation to make the desired concentration, 1 ml of the diluted sample was added to 9 ml of the microorganism dilution solution and mixed well. In the control group, erythromycin, an antibiotic mainly used to suppress acne bacteria, was used. After standing at 37 ° C. for 30 minutes to 1 hour (sometimes mixed), 100 μl of the medium was smeared and then cultured in each culture condition, and the number of colonies was measured after the incubation.

As shown in Figure 2 at 10 ㎍ / ㎖ seems to have a more antimicrobial effect than Erythromycin.

The effect of the present invention is that the compound of the present invention can penetrate into the stratum corneum more quickly and effectively with superior chemical stability and safety and improved lipid-friendly properties than conventional retinoic acid. When it reaches the stratum corneum, the compound releases retinoic acid by various enzymes in the dermis. Eventually, the compounds of the present invention allow effective penetration of retinoic acid into the skin. On the other hand, not only retinoic acid but N-acetyl phytosphingosine is produced by enzyme hydrolysis. Therefore, the N-acetylphytosphingosine is expected to additionally expect its bioactive characteristics as a regulatory substance for epidermal cell growth and differentiation.

Claims (4)

N-acetyl phytosphingosine derivative of the following general formula 2 as a starting material; (Formula 2) In the formula, X means hydrogen or leaving group. Reacting with a retinoic acid derivative of the following general formula (3) in the presence of a condensing agent selected from 1,3-dicyclohexylcarbodiimide (DCC) or N, N-carbonyldiimidazole (CDI) and an organic amine catalyst; (Formula 3) HO-Q In the above formula, Q means a retinoic acid moiety of the following structural formulas (Q-1, Q-2, Q-3). (Q-1) (Q-2) (Q-3) To prepare an N-acetylphytosphingosine retinamide derivative of the following general formula (4); (Formula 4) This is reacted with acetic anhydride again to prepare an acetylated N-acetylphytosphingosine retinamide derivative of the general formula (1) (Formula 1) Acetylated N-acetylphytosphingosine retinamide derivative of claim 1 The acetylated N-acetylphytosphingosine retinamide derivative of claim 2 is used as a functional ingredient in medicine or cosmetics for the purpose of preventing photoaging such as wrinkles, roughness of the skin, moisturizing skin and spots, and for atopic, acne, psoriasis, dry skin, antibacterial and anti-inflammatory. How to Cosmetics, such as cosmetic creams, essences, packs and skins, containing 0.01 to 10% by weight of the acetylated N-acetylphytosphingosine retinamide derivative of claim 2