KR101128450B1 - Composition comprising ß-cyclodextrin derivatives as a stabilizing agent - Google Patents

Abstract

The present invention provides a stabilized composition comprising ascorbic acid, retinol, or epigallocatechin gallate as an active ingredient, and a β-cyclodextrin derivative as a stabilizer. The β-cyclodextrin derivative may be a β-cyclodextrin or a derivative thereof (eg, 6-O-p-tosyl-β-cyclodextrin) and a compound of Formula 1; A compound of formula 2; Or reaction products with antioxidants (eg, dithiothreitol, retinol, silybin, thioglycerol, cysteine, α-lipoic acid, glutathione, etc.).

<Formula 1>

<Formula 2>

In the formula, R ₁ and R ₂ are independently C ₁ ~ C ₁₅ alkyl each other.

β-cyclodextrin, antioxidant

Description

Composition comprising β-cyclodextrin derivatives as a stabilizing agent

The present invention relates to a stabilized composition comprising ascorbic acid, retinol, or epigallocatechin gallate as an active ingredient and a β-cyclodextrin derivative as a stabilizer.

Pharmaceutical compositions and / or cosmetic compositions include various antioxidants. The antioxidants include ascorbic acid, retinol, epigallocatechin gallate, dithiothreitol (DTT), thioglycerol, cysteine, retinol, silybin, α-lipoic acid Various compounds, such as (alpha-lipoic acid) and glutathione, are known.

Ascorbic acid, which is known as a representative antioxidant, has a structure similar to γ-lactone, which has a problem of being easily decomposed by being sensitive to air, particularly oxygen, heat, light, and the external environment. As a solution to the problem of ascorbic acid stability, oxidation of ascorbic acid can be achieved by adding antioxidants, stabilizing in multiple emulsions, stabilizing in oil-based emulsions, and using zinc sulfate and L-tyrosine together. Inhibitors have been proposed (US Pat. No. 4,938,969, EP 553,667 B1, etc.). In addition, to improve the stability of ascorbic acid, sodium ascorbylphosphate, sodium ascorbyl phosphate, magnesium ascorbylphosphate, calcium ascorbylphosphate, and ascorbic acid polypeptide , Ethyl ascorbyl ether, ascorbyl dipalmitate, ascorbyl palmitate, ascorbyl glucoside, ascorbyl glucoside, ascorbyl ethylsilanol pectinate Examples of modifications with derivatives such as Ascorbyl ethylsilanol pectinate have been reported. However, even when the chemical structure of ascorbic acid is modified as described above, when exposed to water, light, or air, there is a limit in securing satisfactory stability, especially in an aqueous medium.

Therefore, there is a need in the art to develop a method that can solve the problems of ascorbic acid instability and discoloration.

Cyclodextrin (CD) is a non-reducing maltoligosaccharide consisting of α-1,4 bonds in which D-glucose (D-glucose) molecules are 6-12 rings and have a donut-shaped cylindrical structure. have. The large inlet of the cyclodextrin cylindrical structure has C2-OH and C3-OH secondary hydroxyl groups, and the narrower inlet contains C6-OH primary hydroxyl groups. The exterior of the cylindrical structure is hydrophilic, and the interior of the cylindrical structure is hydrophobic because of the hydrogen atoms located. Accordingly, small hydrophobic compounds are contained in the cyclodextrin to form inclusion compounds or inclusion compounds, which are used to improve the water solubility of hydrophobic compounds in the fields of medicine, food, cosmetics and the like.

Cyclodextrins are known as α-cyclodextrin consisting of six glucose, β-cyclodextrin consisting of seven, γ-cyclodextrin consisting of eight, and various derivatives are known. For example, Korean Patent Registration No. 10-0363325 discloses a method for preparing β-cyclodextrin derivatives in which octenyl amber acid anhydride is introduced into β-cyclodextrin, and the solubility of retinol in which the β-cyclodextrin derivative is a fat-soluble drug. It is disclosed to improve and exhibit high surface activity and emulsion stability.

The inventors have conducted various studies to develop a method that can increase the stability of ascorbic acid, retinol, and epigallocatechin gallate, which is useful as an antioxidant, especially in aqueous media. As a result, the β-cyclodextrin derivatives incorporating alkyl chains or other antioxidants into the β-cyclodextrin via ester or ether linkages not only increase the stability of ascorbic acid, retinol, and epigallocatechin gallate in the composition. However, it has been found that the problem of discoloration and / or malodor can be solved. In particular, when a β-cyclodextrin derivative having an antioxidant such as dithiothreitol is introduced as a stabilizer, it is dissociated in an in vivo environment (for example, gastrointestinal acid having an acidic pH) or in skin tissue to remove two kinds of antioxidants. It was found that the synergy effect by the two antioxidants can be expected.

Accordingly, the present invention aims to provide a stabilized composition comprising a β-cyclodextrin derivative as a stabilizer and comprising ascorbic acid, retinol, and epigallocatechin gallate.

According to one aspect of the invention, ascorbic acid, retinol, or epigallocatechin gallate (Epigallocatechin gallate) as an active ingredient, and as a stabilizer (a) the reaction of the compound of formula 1 and β-cyclodextrin product; (b) a reaction product of a compound of Formula 2 with β-cyclodextrin; (c) the reaction product of 6-O-p-tosyl-β-cyclodextrin with an antioxidant selected from the group consisting of dithiothreitol, retinol, silybin, thioglycerol, and cysteine; Or (d) a reaction product of α-lipoic acid or glutathione with β-cyclodextrin is provided:

In the formula, R ₁ and R ₂ are independently C ₁ ~ C ₁₅ alkyl each other.

In the composition of the present invention, when the stabilizer is a reaction product of the compound of Formula 1 with β-cyclodextrin or a reaction product of the compound of Formula 2 with β-cyclodextrin, the reaction product is present in NaH in dimethylformamide. It may be obtained by reacting the β-cyclodextrin with a compound of Formula 1 or a compound of Formula 2 under.

In addition, the stabilizer may be a reaction product of the antioxidant and β-cyclodextrin. When the stabilizer is a reaction product of 6-Op-tosyl-β-cyclodextrin with an antioxidant selected from the group consisting of dithiothreitol, retinol, silybin, thioglycerol, and cysteine, the reaction product is dimethylform It may be obtained by reacting the antioxidant with 6-Op-tosyl-β-cyclodextrin in the presence of NaH in the amide.

In addition, when the stabilizer is a reaction product of α-lipoic acid or glutathione and β-cyclodextrin, the reaction product may be obtained by reacting β-cyclodextrin with the α-lipoic acid or glutathione in the presence of a coupling agent and a base. have. The coupling agent may be at least one selected from the group consisting of 3-dimethyl-aminopropyl-N-ethyl carbodiimide, dicyclohexyl carbodiimide, diisopropyl carbodiimide, and ethoxy carbonyl chloride. The base may be at least one selected from the group consisting of dimethylaminopyridine, imidazole pyridine, diisopropylethylamine, and triethylamine.

The composition of the present invention may be in the form of a pharmaceutical composition or cosmetic composition.

The present invention has shown that β-cyclodextrin derivatives with an antioxidant, alkyl ester, or alkyl ether group introduced can effectively increase the stability in the aqueous medium of ascorbic acid, retinol, and epigallocatechin gallate. lost. Therefore, even if the composition according to the present invention is stored for a long time, it is possible to minimize the denaturation by the environment, such as temperature, light, oxygen and water. In particular, β-cyclodextrin derivatives into which antioxidants such as dithiothreitol, thioglycerol, cysteine, retinol, silybin, α-lipoic acid and glutathione have been introduced are dissociated in an in vivo environment or in skin tissue to release β-cyclodextrin and antioxidants. Since there may be zero, synergistic effects of antioxidants, for example, whitening, anti-aging, skin protection, anti-wrinkle, skin moisturizing effect can be expected synergistic effect. Therefore, the β-cyclodextrin derivative may be usefully applied as a stabilizer to pharmaceutical compositions and cosmetic compositions, especially aqueous pharmaceutical compositions and cosmetic compositions.

The present invention includes ascorbic acid, retinol, or epigallocatechin gallate as an active ingredient, and as a stabilizer (a) a reaction product of a compound of Formula 1 with β-cyclodextrin; (b) a reaction product of a compound of Formula 2 with β-cyclodextrin; (c) the reaction product of 6-O-p-tosyl-β-cyclodextrin with an antioxidant selected from the group consisting of dithiothreitol, retinol, silybin, thioglycerol, and cysteine; Or (d) a reaction product of α-lipoic acid or glutathione with β-cyclodextrin, wherein:

<Formula 1>

<Formula 2>

In the formula, R ₁ and R ₂ are independently C ₁ ~ C ₁₅ alkyl each other.

The composition according to the present invention can greatly improve the stability of ascorbic acid, retinol, or epigallocatechin gallate by using the β-cyclodextrin derivatives of (a) to (d) as stabilizers.

In the composition of the present invention, the stabilizer is a reaction product of the compound of formula 1 with β-cyclodextrin [ie (a)] or a reaction product of the compound of formula 2 with β-cyclodextrin [ie, (b )]. The reaction product of the compound of Formula 1 and β-cyclodextrin is at least one of Ra, Rb, and Rc in the formula of Formula 3 is-(CO) R ₁ and the remainder is hydrogen (for example, Ra, Rb When one of Rc is-(CO) R ₁ , the remaining two substituents are all hydrogen, and when two of Ra, Rb, and Rc are-(CO) R ₁ , the other substituent is hydrogen; Ra, Rb, and Rc are all-(CO) R ₁ ), and the reaction product of the compound of Formula 2 with β-cyclodextrin is represented by one or more of Ra, Rb, and Rc in the formula of Formula 3 below -R ₂ and the remainder are hydrogen (e.g., if one of Ra, Rb, and Rc is -R ₂ , the remaining two substituents are both hydrogen; and if two of Ra, Rb, and Rc are -R ₂ , The other substituent is hydrogen; or Ra, Rb, and Rc are all -R ₂ ).

The compound of Formula 1 may be prepared by reacting an alkylcarboxylic acid (ie, C ₁ to C ₁₄ alkylcarboxylic acid) with ethoxycarbonyl chloride in the presence of a base such as triethylamine. In addition, the compound of Formula 2 may be prepared by reacting an alkanol (ie, C ₁ to C ₁₅ alkanol) with methanesulfonyl chloride in the presence of a base such as triethylamine. The reactions may be carried out in an organic solvent such as dichloromethane, and the products may be separated by extraction, concentration under reduced pressure and the like according to a conventional method.

The reaction product of the compound of Formula 1 or the compound of Formula 2 with β-cyclodextrin is dissolved in a solvent such as dimethylformamide and a base such as NaH, and then, the compound of Formula 1 or Formula 2 It can be obtained by reacting the compound. The amount of the compound of Formula 1 or the compound of Formula 2 may be in the range of 1 to 21 equivalents, more preferably 1 to 3 equivalents based on 1 equivalent of β-cyclodextrin. The amount of the base such as NaH may be in the range of 1 to 5 equivalents, more preferably 2 to 3.5 equivalents, based on 1 equivalent of β-cyclodextrin. The reaction of the compound of Formula 1 or the compound of Formula 2 with β-cyclodextrin can be terminated by adding water, and the reaction product obtained is concentrated according to a conventional method, and a process such as precipitation using acetone or the like is performed. Can be separated.

According to the present invention, β-cyclodextrin derivatives into which antioxidants such as dithiothreitol, thioglycerol, cysteine, retinol, silibin, α-lipoic acid and glutathione have been introduced are ascorbic acid, retinol, or epigallocatechin gallate. In addition to improving the stability of, it has been found that it can be dissociated in the in vivo environment or in the skin tissue and be present as β-cyclodextrin and an antioxidant, and thus a synergy effect of the antioxidant can be expected.

When the stabilizer is a β-cyclodextrin derivative introduced with an antioxidant selected from the group consisting of dithiothreitol, retinol, silybin, thioglycerol, and cysteine (that is, an antioxidant having no carboxylic acid group at the end) [ That is, (c)] may be obtained by activating the β-cyclodextrin in the form of 6-Op-tosyl-β-cyclodextrin and reacting with the antioxidant.

Said 6-O-p-tosyl-β-cyclodextrin is known from β-cyclodextrin, for example, Organic Syntheses, Coll. vol. 10. p.686, (2004).

The reaction product of 6-Op-tosyl-β-cyclodextrin with an antioxidant selected from the group consisting of dithiothreitol, retinol, silybin, thioglycerol, and cysteine is 6-Op-tosyl-β-cyclodextrin. It can be obtained by dissolving in a solvent such as dimethylformamide, adding a base such as NaH, and then reacting the antioxidant. The amount of the antioxidant may be in the range of 1 to 1.5 equivalents, more preferably 1 to 1.2 equivalents, relative to 1 equivalent of 6-O-p-tosyl-β-cyclodextrin. The amount of the base such as NaH may be in the range of 1 to 1.5 equivalents, more preferably 1 to 1.2 equivalents, based on 1 equivalent of 6-O-p-tosyl-β-cyclodextrin. The reaction can be carried out in dimethylformamide and can be carried out at room temperature (about 25 ° C.) and under atmospheric pressure. The reaction of the antioxidant with 6-Op-tosyl-β-cyclodextrin can be terminated by adding water, and the reaction product obtained is concentrated according to a conventional method, through a process such as precipitation using acetone or the like. Can be separated.

When the stabilizer is a β-cyclodextrin derivative in which an antioxidant having a carboxylic acid group is introduced at the end, such as α-lipoic acid or glutathione (that is, (d)), the α-lipoic acid or glutathione and β-cyclodextrin The reaction product can be obtained by dissolving β-cyclodextrin in a solvent such as dimethylformamide and reacting α-lipoic acid or glutathione in the presence of a coupling agent and a base. The coupling agent may preferably use 3-dimethyl-aminopropyl-N-ethyl carbodiimide, dicyclohexyl carbodiimide, diisopropyl carbodiimide, ethoxy carbonyl chloride, and the like. Aminopyridine, imidazole pyridine, diisopropylethylamine, triethylamine, etc. can be used preferably. The amount of the antioxidant (α-lipoic acid or glutathione) may be in the range of 1 to 1.5 equivalents based on 1 equivalent of β-cyclodextrin. In addition, the amount of the coupling agent and the base may be in the range of 1 to 1.5 equivalents and 1 to 1.5 equivalents, more preferably 1 to 1.2 equivalents and 1 to 1.2 equivalents, based on 1 equivalent of β-cyclodextrin, respectively. The reaction can be carried out in dimethylformamide, chloroform, or a mixed solvent thereof, and can be carried out at room temperature (about 25 ° C.) and under atmospheric pressure. The reaction product obtained may be concentrated according to a conventional method and separated through a process such as precipitation using acetone or the like.

The composition of the present invention comprises ascorbic acid as an active ingredient and preferably comprises a stabilizer as described above (ie, a stabilizer of (a), (b), (c), or (d)). It may be a pharmaceutical composition or cosmetic composition. The content of the stabilizer may be in the range of 0.5 to 50% by weight based on the total weight of the composition.

The pharmaceutical composition according to the present invention may be in oral or parenteral form and may contain additives such as excipients (or diluents), disintegrants and the like commonly used in the pharmaceutical field. In addition, the content of ascorbic acid contained in the obtained pharmaceutical composition may be varied in consideration of the dosage of the antioxidant.

The cosmetic composition according to the present invention may be preferably an aqueous cosmetic composition, preferably an o / w type cosmetic composition. The cosmetic composition may include carriers commonly used in the field of cosmetic compositions, for example, oils such as liquid paraffin, surfactants such as polysorbate, and the like, fragrances, and preservatives. The cosmetic composition may be prepared for use as a skin, lotion, cream, foundation or essence.

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples. However, these experimental examples and examples are for illustrating the present invention and should not be construed as limiting the present invention.

Preparation Example 1 Preparation of Ethoxy Propionyl Esters

3 g (40.5 mmol.) Propionyl acid and 4.92 g (48.6 mmol.) Triethylamine were added to 30 ml of dichloromethane. 4.83 g (44.55 mmol.) Of ethoxy carbonyl chloride was slowly added dropwise to the reaction solution at -10 to -5 deg. C over 5 minutes. The reaction mixture was stirred at room temperature again for 1 hour and then extracted with 50 ml of 1M HCl aqueous solution. The organic layer was washed with saturated brine, dried over magnesium sulfate (MgSO ₄ ) and filtered. The obtained filtrate was concentrated under reduced pressure to obtain 5.39 g (yield 91%) of ethoxy propionyl ester.

¹ H NMR: (400 MHz, CDCl ₃ ), Chemical shift; 4.20 (2H, q), 2.27 (2H, q), 1.30 (3H, t), 1.14 (3H, t)

Preparation Example 2-10

Instead of propionyl acid, butyryl acid, pentanoyl acid, hexanoyl acid, heptanoyl acid, octanoyl acid, nonannoyl acid, decanoyl acid, undecanoyl acid, and dodecanoyl acid, respectively, preparation examples The reaction was carried out in the same manner as in 1, whereby ethoxy butyryl ester (Preparation Example 2), ethoxy pentanoyl ester (Preparation Example 3), ethoxy hexanoyl ester (Preparation Example 3), ethoxy heptanoyl ester (Preparation Example 5), ethoxy octanoyl ester (Preparation Example 6), ethoxy nonanonoyl ester (Preparation Example 7), ethoxy decanoyl ester (Preparation Example 8), ethoxy undecanoyl ester (Preparation Example 9), and Ethoxy dudecanoyl ester (Preparation 10) was prepared.

Preparation Example 11 Preparation of Propyl Methanesulfonate

3.00 g (49.92 mmol) of propanol and 6.06 g (59.90 mmol) of triethylamine were added to 40 ml of dichloromethane. 6.29 g (54.91 mmol) of methanesulfonyl chloride was slowly added dropwise to the reaction solution at 0 ° C. over 5 minutes with stirring. The reaction mixture was stirred at room temperature again for 1 hour, and then extracted with 50 ml of 1M HCl aqueous solution. The organic layer was washed with saturated brine, dried over magnesium sulfate (MgSO ₄ ) and filtered. The resulting filtrate was concentrated under reduced pressure to give 6.42 g (yield 93%) of propyl methanesulfonate.

¹ H NMR: (400 MHz, CDCl ₃ ), Chemical shift; 3.53 (2H, t), 2.94 (3H, s), 1.52 (2H, m), 0.96 (3H, t)

Preparation Example 12-20

Instead of propanol, n-butanol, n-pentanol, n-hexanol, n-heptanol, n-octanol, n-nonanol, n-decanol, n-undecanol, and n-dudecanol, respectively Reaction was carried out in the same manner as in Production Example 11, to prepare n-butyl methanesulfonate (Preparation Example 12), n-pentyl methanesulfonate (Preparation Example 13), and n-hexyl methanesulfonate (Preparation Example 13). , n-heptyl methanesulfonate (Preparation 15), n-octyl methanesulfonate (Preparation 16), n-nonyl methanesulfonate (Preparation 17), n-decanyl methanesulfonate (Preparation 18), n-Undecanyl methanesulfonate (Preparation Example 19), and n-dudecanyl methanesulfonate (Preparation Example 20) were prepared.

Example 1: Preparation of β-cyclodextrin O-propyl ester

The β-cyclodextrin was washed with acetone and then dried in a vacuum dried oven at 100 ° C. for 5 hours and stored in a desiccator. The dimethylformamide was purified by distillation under reduced pressure, and then it was sealed and stored with pre-dried molecular sieves (linde type 4A molecular sieves).

5.0 g (8.8 mmol) of β-cyclodextrin was added to 90 mL of dimethylformamide prepared above, followed by stirring at room temperature for about 3 hours. 0.742 g of NaH (30.8 mmol) was added thereto, followed by stirring at 25 ° C. for 6 hours. 4.48 g (30.8 mmol) of the ethoxy propionyl ester prepared in Preparation Example 1 was added to the reaction mixture, followed by stirring at 25 ° C. for 3 hours. 10 mL of water was added to the reaction mixture to terminate the reaction, and the resultant was concentrated in a syrup state using a rotary vacuum distillation apparatus. The above process was repeated three times to remove most of dimethylformamide. The concentrated solution was slowly added dropwise to 400 mL of acetone with stirring to form a precipitate, which was filtered to obtain a precipitate which was washed with acetone. The precipitate generation, filtration, and washing processes were repeated three times. The resulting solid was dried under reduced pressure to prepare 1.81 g of β-cyclodextrin O-propyl ester (yield: 28.7%).

Examples 2-10

Instead of the ethoxy propionyl ester, the ethoxy butyryl ester, ethoxy pentanoyl ester, ethoxy hexanoyl ester, ethoxy heptanoyl ester, ethoxy octanoyl ester, ethoxy nonannoyl ester prepared in Preparation Examples 2 to 10 , Ethoxy decanoyl ester, ethoxy undecanoyl ester, and ethoxy dodecanoyl ester, respectively, in the same manner as in Example 1 to carry out the reaction to beta -cyclodextrin O-butyl ester (Example 2 ), β-cyclodextrin O-pentyl ester (Example 3), β-cyclodextrin O-hexyl ester (Example 4), β-cyclodextrin O-heptyl ester (Example 5), β-cyclodextrin O- Octyl ester (Example 6), β-cyclodextrin O-nonyl ester (Example 7), β-cyclodextrin O-decanyl ester (Example 8), β-cyclodextrin O-Under Yl ester was prepared (Example 9), and β- cyclo dextrin O- two decane-yl ester (Example 10).

Test Example 1 Separation and Purification of β-cyclodextrin O-Alkyl Ester

The β-cyclodextrin O-alkyl esters prepared in Examples 1 to 10 were respectively developed on TLC plates (Silica gel 60F254, Merck, Germany) to confirm the composition. The sample was diluted with distilled water, and then developed using a mixed solution of ethyl acetate: ethanol: water = 6: 3: 2 (v / v / v) as a developing solvent. After development, color development was confirmed by spraying sulfuric acid-ethanol mixed solution (3: 7, v / v) and drying at 105 ° C. Β-cyclodextrin and the ethoxy alkyl esters prepared in Preparation Examples 1 to 10 were used together as a reference material and compared. As a result, the β-cyclodextrin O-alkyl esters prepared in Examples 1 to 10 were mainly formed at R _f = 0.70 on TLC, and were present in a small amount at R _f = 0.61 and R _f = 0.17.

In addition, β-cyclodextrin O-alkyl ester having a composition confirmed by TLC was separated by flash chromatography packed with silica gel. As a developing solvent, a mixed solvent of ethyl acetate: ethanol: water = 6: 3: 2 (v / v / v) was used, and a methanol / distilled water (v / v = 1/1) solution was used to wash the column. Each fraction separated was dried under reduced pressure.

Test Example 2 Analysis of the Molecular Weight and the Degree of Substitution of Alkyl Ester of β-Cyclodextrin O-Alkyl Ester

The degree of substitution (DS) of the alkyl carboxylic acid in the β-cyclodextrin O-alkyl ester was calculated according to the following equation from the integrated area of peak intensity in ¹ H-NMR. One molecule of the alkyl carboxylic acid has three protons equivalent to the methyl (methyl, —CH ₃ ) group of the alkyl group, and one molecule of β-cyclodextrin has seven anmeric protons (H-1).

※ Degree of substitution of alkyl carboxylic acid =

   [Peak area of methyl group (3 hydrogens) in alkyl group / 3]

         / [peak area of 7 anomeric protons in β-cyclodextrin / 7]

(In the above formula, when all the hydroxyl groups in the β-cyclodextrin are substituted, the DS becomes maximum 3)

In addition, the molecular weight of the β-cyclodextrin O-alkyl ester prepared in Examples 1 to 10 was determined according to the following formula.

※ Mw = β-cyclodextrin molecular weight + (alkyl acid × DS × 7)

R _f values, substitution degree (DS) and average molecular weight (Mw) that appeared on TLC are shown in Table 1. In addition, it can be seen that the R _f value and the average molecular weight increase as the substitution degree of the formed product and the molecular weight of the alkyl acid increase (see Table 2).

R _f value of TLC DS ( ¹ H-NMR) Mw (average molecular weight) 0.70 0.79 1,456 0.61 0.54 1,353 0.17 0.23 1,227

R _f value of TLC DS ( ¹ H-NMR) Mw (average molecular weight) Example 1 0.70 0.79 1,456 Example 2 0.71 0.78 1,528 Example 3 0.71 0.74 1,580 Example 4 0.73 0.68 1,610 Example 5 0.74 0.69 1,686 Example 6 0.76 0.66 1,726 Example 7 0.77 0.63 1,717 Example 8 0.77 0.61 1,758 Example 9 0.78 0.60 1,807 Example 10 0.78 0.58 1,841

Examples 11-20

Instead of ethoxy propionyl esters, n-propyl methanesulfonate, n-butyl methanesulfonate, n-pentyl methanesulfonate, n-hexyl methanesulfonate, n-heptyl methanesulfonate prepared in Preparation Examples 11 to 20, Same method as Example 1, using n-octyl methanesulfonate, n-nonyl methanesulfonate, n-decanyl methanesulfonate, n-undecanyl methanesulfonate, and n-dudecanyl methanesulfonate, respectively Reaction was carried out with β-cyclodextrin O-propyl ether (Example 11), β-cyclodextrin O-butyl ether (Example 12), β-cyclodextrin O-pentyl ether (Example 13), β- Cyclodextrin O-hexyl ether (Example 14), β-cyclodextrin O-heptyl ether (Example 15), β-cyclodextrin O-octyl ether (Example 16), β-cyclodextrin O-nonyl ether (Example Example 17), β-cyclodextrin O-decanyl ether ( Example 18), β-cyclodextrin O-undecanyl ether (Example 19), and β-cyclodextrin O-dudecanyl ether (Example 20) were prepared.

Test Example 3: Separation and Purification of β-cyclodextrin O-alkyl ether

Β-cyclodextrin O-alkyl ethers prepared in Examples 1 to 10 were separated and purified in the same manner as in Test Example 1. Β-cyclodextrin as a reference material and the ethoxy alkyl esters prepared in Preparation Examples 11 to 20 were developed and compared together. As a result, β-cyclodextrin O-alkyl ethers prepared in Examples 11 to 20 were mainly formed at R _f = 0.70 on TLC, and were present in a small amount at R _f = 0.61 and R _f = 0.17.

Test Example 4: Analysis of the molecular weight of the β-cyclodextrin O-alkyl ether and the degree of substitution of the alkyl ether

The degree of substitution (DS) and the molecular weight of the alkyl ether (confirmation required) in the β-cyclodextrin O-alkyl ether were determined in the same manner as in Test Example 2.

R _f value, substitution degree (DS) and average molecular weight (Mw) that appeared on TLC are shown in Table 3. In addition, it can be seen that the R _f value and the average molecular weight increase as the molecular weight of the alkyl methanesulfonate increases.

R _f value of TLC DS ( ¹ H-NMR) Mw (average molecular weight) Example 11 0.70 0.79 1,367 Example 12 0.71 0.78 1,441 Example 13 0.71 0.74 1,498 Example 14 0.73 0.68 1,535 Example 15 0.74 0.69 1,608 Example 16 0.76 0.66 1,652 Example 17 0.77 0.63 1,717 Example 18 0.77 0.61 1,733 Example 19 0.78 0.60 1,782 Example 20 0.78 0.58 1,817

Preparation Example 21 Preparation of 6-O-p-tosyl-β-cyclodextrin

Organic Syntheses, Coll. vol. 10. According to p.686, 2004, 6-O-p-tosyl-β-cyclodextrin was prepared. 2.5 g of NaOH was dissolved in 150 mL of distilled water, and 5.0 g (4.4 mmol) of β-cyclodextrin hydrate was added and cooled to 0˜5 ° C. 2.0 g (10.5 mmol) of p-toluenesulfonyl chloride was added to the solution, and the mixture was stirred vigorously for 2 hours. Then, 3.0 g (15.7 mmol) of p-toluenesulfonyl chloride was added thereto and maintained at 0 to 5 ° C. Stir for hours. The solution was filtered through celite to remove unreacted materials, and the filtered reaction solution was adjusted to pH 6-7 by dropwise addition of 5M HCl (aq.), And then stored at 0 ° C. overnight to crystallize. The resulting solid was filtered and dried to give 1.87 g of 6-O-p-tosyl-β-cyclodextrin. (Yield 30.9%)

Example 21 Preparation of β-cyclodextrin O-dithiothritol

1.0 g (6.5 mmol) of dithiothreitol was dissolved in 90 mL of dimethylformamide, and 0.39 g (9.76 mmol) of NaH (60% dispersion in mineral oil) was slowly added at 0-5 ° C. and stirred for 15 minutes. . 7.52 g (5.8 mmol) of 6-O-p-tosyl-β-cyclodextrin was slowly added to the solution, followed by stirring at room temperature overnight. 10 mL of water was added after completion | finish of reaction, and it concentrated in syrup state using the rotary distillation apparatus. The water addition and concentration process was repeated three times to remove most of the dimethylformamide. 30 mL of distilled water was added to the concentrate, and the mixture was slowly added dropwise to 400 mL of acetone, followed by three times of precipitation and washing of the reactant, followed by drying under reduced pressure, followed by drying under reduced pressure, 4.2 g of β-cyclodextrin O-dithiothitol. (Yield 51.1%).

Examples 22-25

The reaction was carried out in the same manner as in Example 21 using retinol, silybin, thioglycerol, and cysteine instead of dithiolthitol, and β-cyclodextrin O-retinol (Example 22), β-cyclodextrin O- Silybin (Example 23), β-cyclodextrin O-thioglycerol (Example 24), and β-cyclodextrin O-cysteine (Example 25) were prepared, respectively.

Example 26 Preparation of β-Cyclodextrin O- (α) -Lipoic Acid

5.0 g (8.8 mmol) of β-cyclodextrin and 2.67 g (26.4 mmol) of triethylamine were added to 90 mL of dimethylformamide, and the mixture was stirred at room temperature to dissolve. 2.0 g (9.7 mmol) of α-lipoic acid and 1.9 g (9.7 mmol) of 3-dimethyl-aminopropyl-N-ethyl carbodiimide were slowly added to the solution, followed by 500 mg of N, N'-dimethylaminopyridine. Stirred at room temperature overnight. 10 mL of water was added after completion | finish of reaction, and it concentrated in syrup state using the rotary distillation apparatus. The water addition and concentration process was repeated three times to remove most of the dimethylformamide. 30 mL of distilled water was added to the concentrate, and the mixture was slowly added dropwise to 400 mL of acetone, followed by three times of precipitation and washing of the reactant, followed by drying under reduced pressure, followed by drying under reduced pressure, 3.3 g of β-cyclodextrin O- (α) -lipoic acid. (Yield 28.3%).

Example 27 Preparation of β-cyclodextrin O-glutathione

The reaction was carried out in the same manner as in Example 26 using glutathione instead of α-lipoic acid, to obtain 3.23 g of β-cyclodextrin O-glutathione (yield: 25.8%).

Test Example 5: Analysis of molecular weight and degree of substitution of β-cyclodextrin O-antioxidant

The degree of substitution (DS) and the molecular weight of the antioxidant (need to confirm) in the β-cyclodextrin O-antioxidants prepared in Examples 21 to 25 were determined in the same manner as in Test Example 2, and the results are shown in Table Same as 4.

R _f value of TLC DS ( ¹ H-NMR) Mw (average molecular weight) Example 21 0.72 0.67 1,567 Example 22 0.73 0.65 2,366 Example 23 0.74 0.58 3,029 Example 24 0.69 0.62 1,604 Example 25 0.72 0.67 1,567 Example 26 0.73 0.59 1,987 Example 27 0.65 0.47 2,145

Test Example 6: Evaluation of stability of the composition

Using the β-cyclodextrin derivatives prepared in Examples 1 to 27, respectively, o / w cream bases were prepared with the composition of Table 5 below. The cream base was placed in an oven at a temperature of 45 ° C. to measure the residual content (titer) of ascorbic acid for 4 weeks, thereby performing stability increase and sensory test.

o / w cream base
Test sample (wt%) Control sample (wt%) a b c a b c Stearic Acid Monoglycerin 2.5 2.5 2.5 2.5 2.5 2.5 Liquid paraffin 4.0 4.0 4.0 4.0 4.0 4.0 Polysorbate 1.6 1.6 1.6 1.6 1.6 1.6 Schalen 5.0 5.0 5.0 5.0 5.0 5.0 Carbomer 0.1 0.1 0.1 0.1 0.1 0.1 Hyaluronic acid 0.3 0.3 0.3 0.3 0.3 0.3 Lysine 0.3 0.3 0.3 0.3 0.3 0.3 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 1,3-butylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 Β-CD derivative of Example 1-27 2 2 2 2 (β-CD) 2 (β-CD) 2 (β-CD) Ascorbic acid 6.0 - - 6.0 - - Retinol - 6.0 - - 6.0 - Epigallocatechin gallate - - 6.0 - - 6.0 Spices 0.001 0.001 0.001 0.001 0.001 0.001 antiseptic 0.3 0.3 0.3 0.3 0.3 0.3 Purified water 70.9 70.9 70.9 70.9 70.9 70.9 total 100.0 100.0 100.0 100.0 100.0 100.0

The contents of ascorbic acid, retinol, and epigallocatechin gallate in each o / w cream base were measured three times by high performance liquid chromatography and the average value was calculated. The high performance liquid chromatography conditions are as follows. Ascorbic acid: column-ACE 5-C18 (2.1 * 150 mm, 5 μm), mobile phase-mixed solution of methanol and 0.1% aqueous acetic acid solution (5:95), detector wavelength-UV 240 nm, flow rate-1 ml / min, injection amount-2 μl; Retinol: Column-ACE 5-C18 (4.6 * 150 mm, 5 μm), Mobile phase-Mixture of 90% aqueous methanol solution (90:10), Detector wavelength-UV 325 nm, Flow rate-1 ml / min, Injection volume 2 μl; EGCG: column-ACE 5-C18 (2.1 * 150 mm, 5 μm), mobile phase-acetonitrile / acetic acid / methanol / DW (150: 5: 20: 862), detector wavelength-UV 280 nm, flow rate )-1 ml / min, injection volume-2 μl. As a control, the stability test was performed by adding β-CD instead of O-substituted β-CD.

The sensory test was performed as follows: Each sample was measured by measuring the intensity of the degree of incidence in 10 healthy women in their late 20s, the intensity of the degree was set as shown in Table 6.

Bad smell Explanation 0 Odorless One Detection 2 Normal odor 3 Strong odor 4 Extreme odor 5 Unbearable odor

As described above, the results of performing the residual content and sensory test of ascorbic acid are shown in Tables 7a to 7d below. In Tables 7a to 7d, 'a' refers to ascorbic acid, 'b' refers to retinol, and 'c' refers to epigallocatechin gallate.

From the results of Tables 7a to 7d above, when ascorbic acid, which is easily discolored and / or odorized, is prepared according to the present invention with the addition of O-substituted β-cyclodextrin to prepare an o / w cream base, the overall minimum in aqueous medium It can be seen that high stability is maintained by maintaining a high residual content of more than 85%, and also rarely occurs an odor problem.

Claims (8)

delete As an active ingredient, ascorbic acid, retinol, or epigallocatechin gallate (Epigallocatechin gallate), and as a stabilizer the reaction product of the compound of formula 1 and β-cyclodextrin or the compound of formula 2 and β-cyclodextrin A stabilized composition comprising a reaction product, wherein the reaction product is obtained by reacting a β-cyclodextrin with a compound of Formula 1 or a compound of Formula 2 in the presence of NaH in dimethylformamide: <Formula 1>

<Formula 2>

In the formula, R ₁ and R ₂ are independently C ₁ ~ C ₁₅ alkyl each other. As an active ingredient includes ascorbic acid, retinol, or epigallocatechin gallate (Epigallocatechin gallate), and as stabilizer antioxidant selected from the group consisting of dithiothreitol, retinol, silybin, thioglycerol, and cysteine A stabilized composition comprising a reaction product with 6-Op-tosyl-β-cyclodextrin, the reaction product reacting the antioxidant with 6-Op-tosyl-β-cyclodextrin in the presence of NaH in dimethylformamide. Stabilized composition, characterized in that obtained: <Formula 1>

<Formula 2>

In the formula, R ₁ and R ₂ are independently C ₁ ~ C ₁₅ alkyl each other. delete delete delete delete The composition according to claim 2 or 3, wherein the composition is a cosmetic composition.