KR100437102B1 - New retinol derivatives, the method of preparations and the uses thereof - Google Patents

Abstract

PURPOSE: Provided are a novel retinol derivative, its preparation method in higher yield, and its use. The novel retinol has excellent light-stability, and shows high reactivity to retinoic acid receptor α, while showing low reactivity to retinoic receptor β and γ. It can applied to medical products, cosmetics, soap, shampoo, functional foods, etc., for the prevention and improvement of skin aging. CONSTITUTION: The novel retinol derivative is characterized by carboester bond of a peptide material having COOH group, wherein the peptide material having COOH group is selected from -di, -tri, -poly peptide including N-L- α-aspartyl-L-phenylalanine 1-methylester(AMP;aspartame), N-protection group-aspartame, neotame and the like. Its manufacturing method comprises the steps of: reacting retinylacetate with methanolic solvent and inorganic slat at 25-40 deg.C in a dark room then extracting the reaction product with ether solvent; removing the solvent then followed by mixing a compound having OH group, natural or separated and purified retinol, diethylazodicarboxylate and triphenylphosphate with methylenechloride solvent, and reacting them at room temperature to obtain the ester derivative of retinol; and performing chromatography with reverse-phase, Merck Silicagel 60 RP 18(40-63) micro meter to separate pure ester derivative of retinol.

Description

New retinol derivatives, the method of preparations and the uses

The present invention relates to novel retinol derivatives, their preparation and their use.

Retinol derivatives are important for the control of fetal development, homeostasis, vision, morphogenesis, skin aging, and cell differentiation in animals. In addition, retinol derivatives inhibit viruses and other factors by inhibiting indiscriminate proliferation of cells and inducing differentiation or natural death. Its use has been highlighted as a drug for inhibiting or treating cancer caused by cancer.

In addition, skin aging can be inhibited by maintaining skin cell activity and indirectly interfering with the transmission of ultraviolet rays, and differentiation into muscle nerve cells is regulated according to retinol concentration in hepatocyte differentiation. Therefore, retinol is widely applied in various fields such as medicines and cosmetics as well as derivatives.

U.S. Patent Nos. 4,035,425, 4,064,183 and 4,092,366 describe methods for preparing retinol via several steps. However, pure retinol obtained by such a manufacturing method is unstable to light and is easily photoisomerized and decomposed to have a great influence on the activity of retinol. Therefore, it is generally distributed and used by adding a stabilizer.

U.S. Patent Nos. 4,473,503 and 5,631,244 describe the preparation of retinol derivatives in combination with various carbohydrates to overcome such instability, but the manufacturing steps are complex, uneconomical, and not stable.

Therefore, there is a need for a retinol derivative compound having a simple manufacturing process without being easily decomposed in light or aqueous solution.

The present invention aims to solve the above problems and to provide a stable and novel retinol derivative, an improved production method and a use thereof.

1 is a diagram showing the cytotoxicity analysis of the retinol- (N-formyl-aspartame) of the present invention.

Figure 2 is a diagram showing the RAR / RXR activity regulation analysis of the retinol- (N-formyl-aspartame) of the present invention.

Figure 3 is a view showing the RAR activity regulation analysis of the retinol-mesaconic acid of the present invention.

Figure 4 is a diagram showing the RAR activity regulation analysis of the retinol-fumaric acid of the present invention.

5 is a diagram showing a RAR activity regulation analysis of the retinol-retinol of the present invention.

Figure 6 is a diagram showing the analysis of AP-1 activity regulation of retinol- (N-formyl-aspartame) of the present invention.

The present invention relates to novel retinol derivatives, their preparation and their use.

Retinol derivatives of the invention provide carboester derivatives and ether derivatives of retinol.

The carboester derivatives of retinol of the present invention consist of a -di, -tri, -polypeptide material having a COOH functional group and a retinol carbonate ester bond.

Another carboester derivative of retinol of the present invention consists of an amino acid having a di-COOH functional group and a retinol carboester linkage.

Another carboester derivative of retinol of the present invention consists of a compound having a plurality of double bonds in the COOH functional group and the carbon chain and a retinol carbonate bond.

Another carboester derivative of retinol of the present invention consists of a compound containing a di-COOH functional group and one double bond and a retinol carbonate ester bond.

The ether derivative of retinol of the present invention consists of a compound having an OH functional group and a retinol ether bond.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The structure of the carboester derivative of retinol of the present invention is as follows.

Among the carboester derivatives of the retinol of the present invention, a representative example of a -di, -tri, -polypeptide material having a COOH functional group and a retinol carboester bond may be represented by the structure of Formula 1 below. .

In Formula 1, the peptide material having a COOH functional group is -D, -tree, including NL-α-aspartyl-L-phenylalanine 1-methyl ester (APM; aspartame), N-protecting group-aspartame, neotam, and the like. -Can be selected from polypeptides.

In Formula 1, R ¹ is H, CHO, Retinoic acid (RA), Ac, Boc and the like, R ² is OH, OCH ₃ , OC ₂ H ₅ , Retinol, CLA, etc., n is an integer of 1-6 .

Among the carboester derivatives of the retinol of the present invention, a derivative consisting of an amino acid having a di-COOH functional group and a retinol carboester bond may be represented by the structure of Formula 2 below.

In Formula 2, amino acids having di-COOH include aspartic acid, N-protecting group-aspartic acid, glutamic acid, N-protecting group-glutamic acid, α-aspartyl-L-phenylalanine (α- AP), N-protecting group-α-aspartyl-L-phenylalanine, and the like.

In Formula 2, R ¹ is H, CHO, Retinoic acid (RA), Ac, Boc, etc., R ² is OH, OCH ₃ , OC ₂ H ₅ , Retinol, CLA, etc., n is an integer of 1 to 6 .

Among the carboester derivatives of the retinol of the present invention, a representative example of a compound having a plurality of double bonds in a COOH functional group and a carbon chain and a retinol carbonate ester derivative may be represented by the structure of Formula 3 below.

In Formula 3, a compound having a plurality of double bonds in the COOH functional group and the carbon chain is oleic acid, linolenic acid instead of conjugated linoleic acid (CLA). , Prodlure, leukotrienes and the like.

Among the carboester derivatives of the retinol of the present invention, a derivative comprising a di-COOH functional group and one double bond and a retinol carboester bond may be represented by the structure of the following formula (4).

In Formula 4, the compound containing a di-COOH functional group and one double bond may be selected from the group consisting of maleic acid, fumaric acid, mesaconic acid, and the like.

In Formula 4, R is H, CH ₃ , C ₂ H ₅ .

The ether derivative of the retinol of the present invention is a derivative consisting of a compound having an OH functional group and a retinol ether bond, and a representative example thereof may be represented by the structure represented by the following formula (5).

In Formula 5, the compound having an OH functional group may be selected from the group consisting of alkyl alcohol, allyl alcohol, dienyl alcohol, trienyl alcohol, and the like instead of retinol.

The method for preparing a carboester derivative of retinol of the present invention comprises the steps of 1) reacting retinyl acetate with an inorganic base in a methanolic solvent and then extracting with ether to convert to pure retinol, 2) a peptide containing COOH, di- Combining an amino acid having COOH, a compound having a plurality of double bonds in the carbon chain and a carbon chain, a compound containing a di-COOH function and one double bond with a retinol in the presence of a condensing agent and a catalyst in a solvent, 3 ) Isolating, purifying and recovering the carboester derivative of retinol.

Referring to the method of producing a carboester derivative of the retinol of the present invention step by step.

In the first step, the commercially available retinyl acetate is reacted with methanolic solvent and inorganic base at 25 ~ 40 ° C in a light-blocked dark room to prevent photoisomerization that may occur during the reaction. Extract. In this case, the amount of the inorganic base is about 1 to 3 equivalents, and the extraction solvent includes ethers such as diethyl ether or tetrahydrofuran (THF). The reason why the ether solvent is used is that in the solvent such as ethyl acetate, the residual inorganic base dissolved in the methanolic solvent and the solvent used for extraction can be converted into the starting material retinyl acetate.

In the second step, a peptide containing a COOH, an amino acid having a di-COOH, a compound having a plurality of double bonds in a COOH functional group and a carbon chain, a compound containing a di-COOH functional group and a single double bond and a condensing agent Reaction in methylene chloride (or organic solvent) to activate acid groups or convert to acyl halides followed by the dropwise addition of pure retinol to prepare the carbonate derivatives of retinol.

Condensing agents include N, N'-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), N, N'-carbonyldiimidazole (CDI), N, N'-sulfuryldiimidazole (SDI), SO ₂ Cl and the like, and catalysts for promoting condensation reactions include N, N'-dimethylaminopyridine (DMAP).

The third step is to separate the carboester derivative of pure retinol, using column chromatography using a special silica gel [Reverse-phase, Merck Silicagel 60 RP 18 (40 ~ 63) ㎛).

The method for preparing an ether derivative of retinol according to the present invention comprises the steps of 1) converting retinyl acetate to pure retinol, 2) combining a compound having an OH functional group with retinol, and 3) separating and purifying the ether derivative of retinol. And recovering.

Step by step of the process for producing an ether derivative of the retinol of the present invention.

In the first step, the commercially available retinyl acetate is reacted with a methanolic solvent and an inorganic base at 25 to 40 ° C. in a light-blocked dark room to prevent photoisomerization that may occur during the reaction. Extract.

In the second step, after removing the solvent, a compound having an OH functional group, natural or separated retinol, diethylazodicarboxylate (DEAD), and triphenylphosphate (Ph ₃ P) are methylene chloride. It is added to a solvent and reacted at room temperature to prepare an ether derivative of retinol.

The third step is to separate the ether derivatives of pure retinol, using column chromatography using a special silica gel [Reverse-phase, Merck Silicagel 60 RP 18 (40 ~ 63) ㎛].

Bases, condensing agents, catalysts and solvents that can be used in the preparation of the retinol derivatives are not limited to those enumerated above, and any conventionally known in the art can be used within the scope that does not adversely affect the reaction. have.

Retinol derivatives according to the present invention can be used in medicines, cosmetics, soaps, shampoos and functional foods for the purpose of preventing and improving skin aging due to wrinkles, skin roughness, dryness, abnormal keratinization.

The pharmaceutical composition comprising the retinol derivative of the present invention as an active ingredient may be used for the prevention or treatment of skin cancer or acne, fine wrinkles due to skin aging, irregular pigmentation, skin roughness, sagging skin, and the administration method is oral, topical, transdermal. Administration can be in conventional manner via the intranasal, nasal, inhaled, intraocular, rectal, intravenous, intraperitoneal, intramuscular, intraarterial, or intradermal routes. As an external preparation for skin, transdermal dosage forms such as lotions, ointments, gels, creams, patches or sprays are preferable.

The dosage may vary depending on the patient's age, weight, type of disease, and degree of disease, but can generally be administered at 0.01 to 80 mg / kg, preferably 1 to 3 times daily.

In addition, the cosmetic composition comprising the retinol derivative of the present invention as an active ingredient may be added to cosmetics for the purpose of preventing or alleviating skin pigmentation abnormalities such as blemishes, freckles, senile plaques, and is specifically limited in the formulation thereof. However, the formulation of the flexible cosmetics, nourishing cosmetics, massage creams, nourishing creams, packs, gels or skin adhesive type cosmetics are preferred.

In addition, the food composition comprising the retinol derivative of the present invention as an active ingredient, in order to prevent or alleviate acne, blemishes, fine wrinkles, irregular pigmentation, skin roughness, sagging skin, various foods, meat, beverages, chocolate, snacks, sweets, pizza , Ramen, other noodles, gums, ice cream, alcoholic beverages, vitamin complexes, dietary supplements and the like can be added.

Hereinafter, the present invention will be described in more detail by the following examples, but the present invention is not limited to these examples.

Example 1a : (2E, 4E, 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl (3S) -4 Preparation of-[(1-benzyl-2-methoxy-2-oxoethyl) amino] -3-formylamino-4-oxobutanoate

N-formyl-aspartame (843.3 mg) was added to a three-necked round bottomed flask filled with nitrogen, and 10 ml of methylene chloride and a minimum amount of dimethylformamide (DMF) were added thereto. The reaction mixture was lowered to 0 ° C. and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (683 mg) was slowly added dropwise and stirred for about 30 minutes. Retinol (500 mg) was added to the reaction mixture, and a small amount of N, N'-dimethylaminopyridine (DMAP) was added immediately, followed by stirring for about 1 to 3 hours. After completion of the reaction, the solvent was removed under reduced pressure, dissolved in ethyl acetate and washed several times with water and brine. The organic layer was dried over anhydrous magnesium sulfate (MgSO ₄ ), concentrated at reduced pressure, and separated by column chromatography (Revers-phase).

Yield: 82%

NMR data: 8.81 (s, 1H), 7.12-7.13 (t, 3H, J = 7.08 Hz), 7.04 (d, 2H, J = 7.81 Hz), 6.64 (dd, 1H, J = 13.18 Hz), 6.27 ( d, 1H, J = 15.14 Hz), 6.18 (d, 1H, J = 16.1 Hz), 6.11 (d, 1H, J = 11.72 Hz), 6.07 (d, J = 7.32 Hz), 5.56 (t, J = 7.32 ㎐), 5.2 (s, 1H), 4.75 (d, J = 7.32 ㎐), 4.12 (t, J = 7.32 ㎐), 3.70 (s, 3H), 3.04 (d, J = 6.84 ㎐), 2.94 ( dd, 2H, J = 4.39 ㎐, 2.65 (d, 2H, J = 6.84 ㎐), 2.01 (t, 2H, J = 5.86), 1.95 (s, 3H), 1.88 (s, 3H), 1.17 (s , 3H), 1.62 (m, 2H), 1.46 (m, 2H), 1.02 (s, 6H)

Example 1b: another exemplary embodiment of Example 1a

Only the condensing agent used N, N'-carbonyldiimidazole (CDI), and prepared in the same manner as in Example 1a.

Example 1c: Other exemplary embodiments of the Example 1a

Only the condensing agent used SO ₂ Cl, it was prepared in the same manner as in Example 1a.

Example 1d: other exemplary embodiments of the Example 1a

Only the condensing agent used N, N'-sulfuryldiimidazole (SDI), and prepared in the same manner as in Example 1a.

Example 2: (2E, 4E, 6E, 8E) dimethyl -3,7-9--(2,6,6-trimethyl 1--cyclohexenyl) -2,4,6,8-nonatetraenyl (3S) -4 -[(1-benzyl-2-methoxy-2-oxoethyl) amino] -3-amino-4-oxobutanoate. Preparation of Hydrochloride

Aspartame hydrochloride (150 mg) was added to a three-neck round bottom flask filled with nitrogen, and 15 ml of methylene chloride and a minimum amount of dimethylformamide (DMF) were added to dissolve it. The reaction mixture was lowered to 0 ° C. and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (128 mg) was slowly added dropwise and stirred for about 30 minutes. Retinol (130 mg) was added to the reaction mixture, and a small amount of N, N'-dimethylaminopyridine (DMAP) was added immediately, followed by stirring for about 1 to 3 hours. After completion of the reaction, the solvent was removed under reduced pressure, dissolved in ethyl acetate and washed several times with water and brine. The organic layer was dried over anhydrous magnesium sulfate, concentrated at reduced pressure, and separated by column chromatography.

Yield: 62%

NMR data: 7.11-7.12 (t, 3H, J = 7.08 Hz), 7.0 (d, 2H, J = 7.81 Hz), 6.61 (dd, 1H, J = 13.18 Hz), 6.20 (d, 1H, J = 15.14 I), 6.10 (d, 1H, J = 16.1 kPa), 6.05 (d, 1H, J = 11.72 kPa), 6.02 (d, J = 7.32 kPa), 5.53 (t, J = 7.32 kPa), 5.10 (bs , 1H), 4.70 (d, J = 7.32 kPa), 4.12 (t, J = 7.32 kPa), 3.70 (s, 3H), 3.14 (d, J = 6.84 kPa), 2.84 (dd, 2H, J = 4.39 I), 2.45 (d, 2H, J = 6.84 Hz), 2.01 (t, 2H, J = 5.86), 1.75 (s, 3H), 1.68 (s, 3H), 1.17 (s, 3H), 1.52 (m , 2H), 1.26 (m, 2H), 1.02 (s, 6H)

Example 3 : Di [(2E, 4E, 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl] (2S Preparation of) -2-[(tert-butoxycarbonyl) amino] butanedioate

N-Boc-aspartic acid (1.2 g) was added to a three-necked round bottom flask filled with nitrogen, and 20 ml of methylene chloride and a minimum amount of dimethylformamide (DMF) were added to dissolve it. The reaction mixture was lowered to 0 ° C. and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (1.36 g) was slowly added dropwise and stirred for about 30 minutes. Retinol (0.51 g) was added to the reaction mixture, followed by immediately adding a small amount of N, N'-dimethylaminopyridine (DMAP), followed by stirring for about 1 to 3 hours. After completion of the reaction, the solvent was removed under reduced pressure, dissolved in ethyl acetate and washed several times with water and brine. The organic layer was dried over anhydrous magnesium sulfate, concentrated at reduced pressure, and separated by column chromatography.

Yield: 53%

NMR data: 6.64 (dd, 3H, J = 13.18 Hz), 6.23 (d, 2H, J = 15.14 Hz), 6.07 (d, 2H, J = 16.11 Hz), 6.05 (d, 2H, J = 11.23 Hz) , 5.54 (t, 2H, J = 6.60 Hz), 4.77 (d, 2H, J = 7.23 Hz), 4.71 (d, 2H, J = 7.32 Hz), 2.97 (dd, 2H, J = 4.39 Hz), 2.01 (t, 4H, J = 5.86 Hz), 1.95 (s, 6H), 1.88 (s, 6H), 1.17 (s, 6H), 1.62 (m, 4H), 1.46 (m, 4H), 1.42 (s, 9H), 1.02 (s, 12H)

Example 4 : Di [(2E, 4E, 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl] (2S Preparation of 2-aminobutanedioate

Aspartic acid (610 mg) and N, N'-dicyclohexylcarbodiimide (DCC) (720 mg) were added to a three-necked round bottom flask filled with nitrogen, and 10 ml of chloroform and a minimum amount of dimethylformamide (DMF) were added. And dissolved. A small amount of N, N'-dimethylaminopyridine (DMAP) was added and retinol (324 mg) was added followed by stirring for about 1 to 3 hours. After completion of the reaction, the solvent was removed under reduced pressure, dissolved in ethyl acetate and washed several times with water and brine. The organic layer was dried over anhydrous magnesium sulfate, concentrated at reduced pressure, and separated by column chromatography.

Yield: 50%

NMR data: 6.54 (dd, 3H, J = 13.18 Hz), 6.08 (d, 2H, J = 15.14 Hz), 6.07 (d, 2H, J = 16.11 Hz), 6.05 (d, 2H, J = 11.23 Hz) , 5.54 (t, 2H, J = 6.60 Hz), 4.57 (d, 2H, J = 7.23 Hz), 4.51 (d, 2H, J = 7.32 Hz), 2.67 (dd, 2H, J = 4.39 Hz), 2.01 (t, 4H, J = 5.86 Hz), 1.95 (s, 6H), 1.88 (s, 6H), 1.17 (s, 6H), 1.62 (m, 4H), 1.46 (m, 4H), 1.02 (s, 12H)

Example 5 : Di [(2E, 4E, 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl] 2- Preparation of (acetylamino) succinate

N-acetyl aspartic acid (610 mg) and N, N'-dicyclohexylcarbodiimide (DCC) (720 mg) were added to a three-necked round bottom flask filled with nitrogen, and 10 ml of chloroform and a minimum amount of dimethylformamide ( DMF) was added to dissolve. A small amount of N, N'-dimethylaminopyridine was added, retinol (324 mg) was added, followed by stirring for about 1 to 3 hours. After completion of the reaction, the solvent was removed under reduced pressure, dissolved in ethyl acetate and washed with water and brine (20 mL × 2). The organic layer was dried over anhydrous magnesium sulfate, concentrated at reduced pressure, and separated by column chromatography.

Yield: 50%

NMR data: 6.54 (dd, 3H, J = 13.18 Hz), 6.08 (d, 2H, J = 15.14 Hz), 6.07 (d, 2H, J = 16.11 Hz), 6.05 (d, 2H, J = 11.23 Hz) , 5.54 (t, 2H, J = 6.60 Hz), 4.57 (d, 2H, J = 7.23 Hz), 4.51 (d, 2H, J = 7.32 Hz), 2.67 (dd, 2H, J = 4.39 Hz), 2.01 (t, 4H, J = 5.86 Hz), 1.95 (s, 6H), 1.88 (s, 6H), 1.17 (s, 6H), 1.62 (m, 4H), 1.46 (m, 4H), 1.42 (s, 9H), 1.02 (s, 12H)

Example 6 : (2E, 4E, 6E, 8E) -3.7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl (9Z, 11E) -9 , 11-octadecadienoate

In a three-necked round bottom flask filled with nitrogen, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (0.71 g, 3.67 mmol) was added to 15 ml of anhydrous methylene chloride, and then dissolved. The temperature was lowered to 0 ° C., and conjugated linoleic acid (CLA) (0.85 g, 3.05 mmol) was added thereto, followed by stirring for 30 minutes. To this reaction mixture, retinol (0.87 g, 3.05 mmol) and N, N'-dimethylaminopyridine catalyst amount were dissolved in 7 ml of anhydrous methylene chloride and slowly added dropwise, followed by stirring at room temperature for 4 hours. The organic layer was washed with water (20 mL × 2) and brine (20 mL × 2), dried over MgSO ₄ , and the solvent was distilled off under reduced pressure and separated by column chromatography (solvent used: ethyl acetate / hexane = 1 //). 8).

Yield: 80%

NMR data (400MHz, CDCl₃): 6.64 (dd, 1H, J = 15.14 Hz, 11.23 Hz), 6.29 (d, 1H, J = 15.14 Hz), 6.17 (d, 1H, J = 16.11 Hz), 6.12 (d, 1H, J = 16.11 Hz), 6.09 (d, 1H, J = 11.23 Hz), 5.96 (m, 1H, J = 10.74 Hz, 5.86 Hz), 5.93 (m, 1H, J = 10.74 Hz), 5.61 (m, 1H, J = 6.83 Hz, 7.32 Hz), 5.32 (t, 1H, J = 7.32 Hz), 5.30 (m, 1H, J = 10.74 ㎐, 6.83 ㎐), 4.72 (d, 2H, J = 7.32 Hz), 2.30 (t, 2H, J = 5.86 Hz), 2.14 (m, 2H, J = 5.86 Hz, 10.74 Hz), 2.08 (m, 2H, J = 5.86 Hz, 7.32 Hz), 2.01 ( m, 2H, J = 5.86 Hz, 1.96 (s, 3H), 1.89 (s, 3H), 1.71 (s, 3H), 1.60 (m, 2H, J = 5.86 Hz), 1.47 (m, 2H, J = 5.86 kPa), 1.29 (m, 16H, J = 5.86 kPa), 1.02 (s, 16H), 0.87 (t, 3H, J = 7.32 kPa)

Example 7: Di [(2E, 4E , 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl] of malate Produce

In a three-necked round bottomed flask filled with nitrogen, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (334.6 mg, 1.75 mmol) was added to 5 ml of anhydrous methylene chloride, and then dissolved. Was lowered to 0 ° C. and maleic acid (78 mg, 0.58 mmol) was added and stirred for 30 minutes. To this reaction mixture, retinol (500 mg, 1.75 mmol) and N, N'-dimethylaminopyridine catalyst amount were dissolved in 6 ml of anhydrous methylene chloride, and slowly added dropwise, and stirred at room temperature for 4 hours. The organic layer was washed with water (20 mL × 2) and brine (20 mL × 2), dried over MgSO ₄ , and the solvent was distilled off under reduced pressure and separated by column chromatography (solvent used: ethyl acetate / hexane = 1 //). 30).

Yield: 48%

NMR data (400MHz, CDCl₃): 6.60 (dd, 2H, J = 15.14 Hz, 11.23 Hz) 6.23 (d, 2H, J = 15.14 Hz), 6.17 (d, 2H, J = 16.11 dB), 6.13 (d, 2H, J = 16.11 dB), 6.07 (d, 2H, J = 11.23 dB), 5.50 (t, 2H, J = 7.32 Hz), 4.73 (d, 2H, J = 7.32 Hz), 4.65 (d, 2H, J = 7.32 Hz), 5.23 (bq, 1H, J = 16.60 Hz), 3.00 (d, 1H, J = 16.60 Hz), 2.90 (d, 1H, J = 16.60 Hz), 1.98 (m, 4H, J = 5.86 Hz), 1.94 (s, 6H), 1.83 (s, 6H), 1.68 (s, 6H), 1.61 (m, 4H, J = 5.86 Hz) , 1.46 (m, 4H, J = 5.86 Hz), 1.02 (s, 12H)

Example 8: Di [(2E, 4E , 6E, 8E) -3,7-dimethyl-9- (2, 6, 6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl] (E ) -2-butenedioate Preparation

In a three-necked round bottomed flask filled with nitrogen, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (1.4 g, 7.32 mmol) was added to 20 ml of anhydrous methylene chloride, and dissolved. It was lowered to 0 ℃, fumaric acid (0.39 g, 3.05 mmol) was added and stirred for 30 minutes. To this reaction mixture, retinol (1.75 g, 6.10 mmol) and N, N'-dimethylaminopyridine catalyst amount were dissolved in 7 ml of anhydrous methylene chloride and slowly added dropwise, followed by stirring at room temperature for 4 hours. The organic layer was washed with water (20 mL × 2) and brine (20 mL × 2), dried over MgSO ₄ , and the solvent was distilled off under reduced pressure and separated by column chromatography (solvent used: ethyl acetate / hexane = 1 //). 30).

Yield: 50%

NMR data: 6.86 (s, 2H), 6.65 (dd, 2H, J = 24 Hz), 6.08 to 6.29 (m, 8H), 5.63 (t, 2H, J = 15 Hz), 4.85 (d, 4H, J) = 15 μs), 1.99 to 2.03 (m, 4H), 1.96 (s, 6H), 1.91 (s, 6H), 1.70 (s, 6H), 1.59 to 1.62 (m, 4H), 1.45 to 1.47 (m, 4H), 1.02 (s, 12H)

Example 9 D i [(2E, 4E, 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2,4,6,8-nonatetraenyl] ( E) -2-methyl-2-butenedioate Preparation

In a three-necked round bottomed flask filled with nitrogen, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride [EDCI] (1.4 g, 7.32 mmol) was added to 20 ml of anhydrous methylene chloride, and dissolved. Was lowered to 0 ° C., and mesaconic acid (0.39 g, 3.05 mmol) was added and stirred for 30 minutes. To this reaction mixture, retinol (1.75 g, 6.10 mmol) and N, N'-dimethylaminopyridine catalyst amount were dissolved in 7 ml of anhydrous methylene chloride and slowly added dropwise, followed by stirring at room temperature for 4 hours. The organic layer was washed with water (20 mL × 2) and brine (20 mL × 2), dried over MgSO ₄ , and the solvent was distilled off under reduced pressure and separated by column chromatography (solvent used: ethyl acetate / hexane = 1 //). 30).

Yield: 53%

NMR data: 6.79 (s, 1H), 6.65 (dd, 2H, J = 27 Hz), 6.08 to 6.29 (m, 8H), 5.63 (t, 2H, J = 14 Hz), 4.83 (dd, 4H, J) = 16 ㎐), 2.29 (s, 3H, J = 4.39 ㎐), 1.99 to 2.03 (m, 4H), 1.96 (s, 6H), 1.91 (s, 6H), 1.67 (s, 6H), 1.58 to 1.62 (m, 4H), 1.45-1.47 (m, 4H), 1.02 (s, 12H)

Example 10 : (1E, 3E, 5E, 7E) -9 [(2E, 4E, 6E, 8E) -3,7-dimethyl-9- (2,6,6-trimethyl-1-cyclohexenyl) -2, Preparation of 4,6,8-nonatetraenyl] oxy-3,7-dimethyl-1- (2,6,6-trimethyl-1-cyclohexenyl) -1,3,5,7-nonateraene

Retinol (0.6 g, 2.10 mmol), DEAD (Diethylazodicarboxylate) (0.3 g, 1.15 mmol) and Ph ₃ P (0.33 g, 1.15 mmol) were added to a three-necked round bottom flask filled with nitrogen, and anhydrous methylene chloride 20 ML was added to dissolve and stirred vigorously at room temperature for 20 minutes. After completion of the reaction, 50 ml of water was added, followed by extraction with ethyl acetate (30 ml × 2). The organic layer was washed with water (50 mL) and brine (50 mL), dried over MgSO ₄ , and the solvent was distilled off under reduced pressure and separated by column chromatography (use solvent: ethyl acetate / hexane = 1/1).

Yield: 84%

NMR data: 6.05 (q, 4H, J = 16.11 Hz), 5.14 (br s, 6H), 4.16 (q, 4H, J = 7.32 Hz), 1.99 (s, 6H), 1.74 (s, 6H), 1.68 (s, 6H), 1.63 (t, 4H, J = 5.86 Hz), 1.45 (m, 4H), 1.01 (s, 6H), 1.00 (s, 6H)

Experimental Example 1 Cytotoxicity Test of Retinol- (N-Formyl-Aspartame)

Cell lines used to analyze cytotoxicity by retinol derivatives are SK-Hep-1 (liver cancer), MDA-MB-231 (breast cancer), HaCAT (skin cancer), HCT116 (colon cancer). These cell lines were maintained and cultured in DMEM (Dulbecco's Modified Eagle's Medium) / 10% FBS containing 10% fetal bovine serum (FBS) (GibcoBRL, Gaithersburg, MD). 100 μl of medium and 3 × 10 ³ cells of the cells were dispensed in a 96 well microtiter plate and cultured for 24 hours. Retinol derivatives of different concentrations (0, 500, 1000, 5000 nM) were treated for 4 days in cultured cell lines, and the extent of proliferation inhibition was measured by counting living cells directly with a hemocytometer.

The retinol derivatives used in the experiment were retinol- (N-formyl-aspartame), retinol, 4-HPR [N- (4-hydroxyphenyl) retinamide] (Sigma Co., St. Louis, MO), respectively dimethyl It was used by dissolving in sulfoxide, the concentration of dimethyl sulfoxide added to the culture was not to exceed 0.01%.

As shown in Figure 1, when treated by concentration, retinol- (N-formyl-aspartame) is completely toxic to SK-Hep-1 (liver cancer), MDA-MB-231 (breast cancer), HaCAT (skin cancer) cell lines. The HCT116 (colon cancer) cell line showed mild toxicity compared to retinol.

Experimental Example 2 : Retinol- (N-formyl-aspartame) retinic acid receptor / retinoid X receptor (RAR / RXR) activity assay

To analyze the effect of retinol- (N-formyl-aspartame) on the activity of the retinic acid receptor, skin cancer (HaCAT) cell line was used to measure the activity on the retinic acid receptor / retinoid ex receptor.

DR5-tk-CAT or DR1-tk-CAT, a recombinant gene consisting of DR1 or DR5, thymine kinase promoter, and CAT (chloramphenicol acetyl transferase) gene Was appropriately combined with plasmid DNA expressing the retinic acid receptors α, β, and γ or the retinoid ex receptors α, β, and γ. Thereafter, the cells were incubated in DMEM / 10% FBS medium for one day, and then each retinol derivative was added to 1 μM and cultured again for 5 days at 37 ° C. under 5% CO ₂ . Cells were washed with phosphate-buffered saline (PBS), protein was extracted from each cell, and then transfection efficiency was determined by measuring β-galactosidase activity and protein quantification. The degree of transcriptional activity was analyzed by CAT ELISA (Roche Molecular Biochemicals, Mannheim, Germany).

To analyze the effect of retinol- (N-formyl-aspartame) on the activity of retinic acid receptors, each retinic acid receptor (α, β, or γ) expression vector and DR5-tk for activity measurement in skin cancer (HaCAT) cell lines. -CAT plasmid was introduced into liposomes. Thereafter, retinol- (N-formyl-aspartame) was treated and cell extracts were obtained to quantify the expression of CAT by CAT ELISA to measure the change in activity of each retinic acid receptor according to retinol- (N-formyl-aspartame).

As shown in FIG. 2, retinol- (N-formyl-aspartame) showed high activity on retinic acid receptor α, similar to retinol, and generally weak activity on retinic acid receptors β and γ (A). Retinol and retinol- (N-formyl-aspartame) derivatives were inactive against the retinoid ex receptors (α, β, γ) (B).

Experimental Example 3 : Retinol-mesaconic acid retinic acid receptor (RAR) activity assay

In order to analyze the effect of retinol-mesaconic acid on the activity of the retinic acid receptor, the Cos-1 cell line was used to measure the activity on the retinic acid receptor.

Recombinant gene DR5-tk-CAT, which consists of DR5, thymine kinase promoter, and CAT (chloramphenicol acetyl transferase) genes, which express retinic acid receptors α, β, and γ Properly combined with plasmid DNA was cotransfected into Cos-1 cell line with lipofectamine (GibcoBRL). Thereafter, the cells were incubated in DMEM / 10% FBS medium for one day, and then each retinol derivative was added to 1 μM and cultured again for 5 days at 37 ° C. under 5% CO ₂ . Cells were washed with phosphate buffer (PBS), protein was extracted from each cell, and transfection efficiency was determined by measuring β-galactosidase activity and protein quantification. Transcriptional activity to RAR receptor was determined by CAT ELISA. Analyzed.

As shown in FIG. 3, retinol-mesaconic acid showed high activity on retinic acid receptor α, similar to retinol, and generally weak activity on retinic acid receptors β and γ.

Experimental Example 4 : Retinol-fumaric acid retinic acid receptor (RAR) activity assay

In order to analyze the effect of the retinol-fumaric acid on the activity of the retinic acid receptor, Cos-1 cell line was used to measure the activity of the retinic acid receptor in the same manner as in Experiment 3.

As shown in FIG. 4, retinol-fumaric acid showed high activity on retinic acid receptor α, similar to retinol, and showed weak overall activity on retinic acid receptors β and γ.

Experimental Example 5 : Retinol-Retinol Retinic Acid Receptor (RAR) Activity Assay

In order to analyze the effect of the retinol-retinol on the activity of the retinic acid receptor, Cos-1 cell line was used to measure the activity of the retinic acid receptor in the same manner as in Experimental Example 3.

As shown in FIG. 5, retinol-retinol showed high activity on retinic acid receptor α like retinol, and generally weak activity on retinic acid receptors β and γ.

Experimental Example 6 : Inhibition of the activity of retinol- (N-formyl-aspartame) activation protein-1 (AP-1)

Since active protein-1 (a component of c-Jun) is a transcription factor that induces the expression of collagen degrading enzyme, which is the main cause of skin wrinkles, the anti-wrinkle effect of the retinol derivative was confirmed through an activity inhibition experiment of active protein-1.

CAT reporter (Coll-CAT) having a collagen promoter containing an active protein-1 reaction site (TRE) was transfected into skin cancer (HaCAT) cell lines, and the activity of the activated protein-1 by retinol derivatives was measured by CAT ELISA. . In some cases, the effects of retinol derivatives on the transcriptional activity of c-Jun were analyzed by cotransfecting c-Jun or retinic acid receptor expression vectors.

Since the prepared retinol- (N-formyl-aspartame) derivatives induced the inhibition of active protein-1, which causes skin wrinkles, it would be useful for cosmetics. Similar to the retinic acid receptor experiment described above, an activated protein-1 (c-Jun) expression vector and a Coll-CAT plasmid for activity measurement were introduced into liposomes into a skin cancer (HaCAT) cell line. After treatment of retinol- (N-formyl-aspartame) derivatives and cell extracts to quantify the expression of CAT by CAT ELISA, the activity change of active protein-1 according to the retinol- (N-formyl-aspartame) derivatives was measured. It was.

As shown in Figure 6, the expression of the collagen degrading enzyme that causes wrinkles when c-Jun is expressed in the cell is increased by about 4.5 times, and when the retinol is treated with the retinic acid receptor α, the expression of the collagenase is about 47%. Retinol was inhibited by about 60% and retinol- (N-formyl-aspartame) by about 44%. Similar inhibition rates could be obtained in other cases (FIG. 6, c-Jun or + c-Jun). Through this, retinol- (N-formyl-aspartame) was found to be inferior to retinic acid but similarly inhibited the activity of active protein-1 similar to retinol.

Experimental Example 7 Experiment of Inhibiting Activity of Retinol-Mesaconic Acid Active Protein-1

After the CAT reporter (Coll-CAT) having a collagen promoter containing the active protein-1 reaction site (TRE) was transfected into the Cos-1 cell line, the activity inhibition experiment of the active protein-1 in the same manner as in Experimental Example 6 Was done.

As a result, the expression of collagen degrading enzyme which causes wrinkles when c-Jun is expressed in cells was 100% .Retinol treatment inhibited the expression of collagen degrading enzyme by about 42%, retinic acid by about 50%, retinol Mesaconic acid was inhibited by about 30%. Through this, retinol-mesaconic acid was found to inhibit the activity of the active protein-1 weakly than retinic acid and retinol.

Experimental Example 8 : Inhibition of the activity of the active protein-1 of retinol-fumaric acid

After the CAT reporter (Coll-CAT) having a collagen promoter containing the active protein-1 reaction site (TRE) was transfected into the Cos-1 cell line, the activity inhibition experiment of the active protein-1 in the same manner as in Experimental Example 6 Was done.

As a result, the expression of collagen degrading enzyme which causes wrinkles when c-Jun is expressed in cells was 100% .Retinol treatment inhibited the expression of collagen degrading enzyme by about 42%, retinic acid by about 50%, retinol Fumaric acid was hardly inhibited.

Experimental Example 9 Experiment of Inhibiting Activity of Retinol-Retinol Active Protein-1

After the CAT reporter (Coll-CAT) having a collagen promoter containing the active protein-1 reaction site (TRE) was transfected into the Cos-1 cell line, the activity inhibition experiment of the active protein-1 in the same manner as in Experimental Example 6 Was done.

As a result, the expression of collagen degrading enzyme that causes wrinkles when c-Jun is expressed in cells was 100%. When retinol was treated, the expression of collagenase was suppressed by about 43%, retinic acid was about 47.5%, retinol Retinol was inhibited by about 41.5%. It was confirmed that retinol-retinol inhibits the activity of active protein-1 similar to retinic acid and retinol.

The retinol derivatives of the present invention have been found to be more stable against light than conventionally known retinol, and exhibit stability without any change even after about 10 days.

Like the retinol, the retinol derivatives of the present invention showed high activity on the retinic acid receptor α, and generally showed weak activity on the retinic acid receptors β and γ. There was no activity for the retinoid ex receptors α, β, and γ.

In addition, the retinol derivatives of the present invention also inhibited the activity of c-Jun to the extent that retinol is similar to that of retinic acid receptors.

Therefore, the retinol derivative of the present invention can be effectively used in medicines, cosmetics, soaps, shampoos and functional foods for the purpose of preventing and improving skin aging due to wrinkles, rough skin, dryness, abnormal keratinization and the like.

Claims (22)

delete -D, -Tree having a COOH functional group comprising any one selected from the group consisting of NL-α-aspartyl-L-phenylalanine 1-methylester (APM; aspartame), N-protecting group-aspartame and neotam; or -A retinol derivative characterized by consisting of a polypeptide and a retinol carbonate ester bond delete As amino acids with di-COOH functional groups, aspartic acid, N-protecting group-aspartic acid, glutamic acid, N-protecting group-glutamic acid, α-aspartyl-L-phenylalanine (α-AP) and Retinol derivative, characterized in that consisting of retinol carbonate ester bonds and amino acids selected from the group consisting of N-protecting group-α-aspartyl-L-phenylalanine delete Retinol derivatives comprising a prodlure having a plurality of double bonds in a COOH functional group and a carbon chain and a retinol carbonate ester bond delete Retinol derivatives comprising maleic acid or mesaconic acid and retinol carbonate bonds delete A compound having an OH function, a retinol derivative comprising a retinol ether bond and a compound selected from the group consisting of retinol, alkyl alcohol, allyl alcohol, dienyl alcohol and trienyl alcohol 1) Retinyl acetate is reacted with an inorganic base in methanolic solvent and then extracted with ether solvent to convert to pure retinol, 2) Peptides containing COOH, amino acids with di-COOH, COOH functional groups and a large number in the carbon chain Combining a compound having a double bond, a di-COOH functional group and a compound containing one double bond with a retinol in a solvent in the presence of a condensing agent and a catalyst, 3) separating, purifying and recovering a retinol carbonate derivative Method for producing a carboester derivative of retinol consisting of 1) reacting retinyl acetate with an inorganic base in a methanolic solvent and then extracting with ether solvent to convert to pure retinol, 2) combining a compound having an OH functional group with retinol, 3) ether derivative of retinol Method for producing an ether derivative of retinol consisting of the steps of separating, purifying and recovering The method for preparing a retinol derivative according to claim 11 or 12, wherein the ether solvent in step 1) is diethyl ether or tetrahydrofuran. The method of claim 11, wherein N, N'-dicyclohexylcarbodiimide (DCC), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), N, N'-carbonyldiimidazole (CDI), N, N'-sulfuryldiimidazole (SDI) and SO ₂ Cl method for the preparation of retinol derivatives A pharmaceutical composition for preventing and treating skin aging containing the retinol derivative of any one of claims 2, 4, 6, 8 and 10. A cosmetic composition for preventing and improving skin aging containing the retinol derivative according to any one of claims 2, 4, 6, 8 and 10. Soap and shampoo composition for preventing and improving skin aging containing the retinol derivative according to any one of claims 2, 4, 6, 8 and 10. A food composition for preventing and improving skin aging containing the retinol derivative according to any one of claims 2, 4, 6, 8 and 10. A pharmaceutical composition for preventing and treating acne containing the retinol derivative of any one of claims 2, 4, 6, 8 and 10. A cosmetic composition for preventing and improving acne containing the retinol derivative according to any one of claims 2, 4, 6, 8 and 10. Soap and shampoo composition for preventing and improving acne containing the retinol derivative of any one of claims 2, 4, 6, 8 and 10. A food composition for preventing and improving acne containing the retinol derivative of any one of claims 2, 4, 6, 8, and 10.