JP6991142B2 - 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid, compositions containing it, and methods for producing them. - Google Patents

Description

The present invention relates to 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid, a composition containing the same, and a method for producing the same.

L-ascorbic acid (vitamin C) is known to have a whitening effect and is useful as a cosmetic material, but since it is unstable and easily oxidized, various L-ascorbic acid derivatives have been researched and developed in the 1990s. 2-O-α-D-glycosyl-L-ascorbic acid (L-ascorbic acid-2-glycoside) has been developed as a stable and decomposed and absorbed derivative in the body (Patent Documents 1 to 3).
2-O-α-D-glucopyranosyl-L-ascorbic acid, which is a typical 2-O-α-D-glycosyl-L-ascorbic acid, has the following chemical formula (2).

It has the chemical structure shown by and is hydrolyzed to produce L-ascorbic acid and D-glucose. 2-O-α-D-glucopyranosyl-L-ascorbic acid is less likely to be oxidized than L-ascorbic acid and has excellent stability, but it is easily hydrolyzed in the body and has the same bioactivity as L-ascorbic acid. Is known (Patent Documents 2 and 3), and is expected to have a wide range of applications such as cosmetics, foods and pharmaceuticals. Further, as an L-ascorbic acid derivative having further improved oil solubility, an acyl group was introduced into the hydroxyl group at the 6-position of the L-ascorbic acid residue in 2-O-α-D-glucopyranosyl-L-ascorbic acid. 6-Acyl-2-O-α-D-glucopyranocil-L-ascorbic acid was developed (Patent Document 4).

Japanese Patent No. 2832848 Japanese Unexamined Patent Publication No. 03-135992 Japanese Unexamined Patent Publication No. 03-183492 Japanese Unexamined Patent Publication No. 11-286497

A 2-O-α-D-glycosyl-L-ascorbic acid derivative or a composition containing the derivative, which is excellent in solubility in water and / or an organic solvent and / or permeability to the skin. The challenge is to provide things.

As a result of diligent research, the present inventors have made 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and / or a composition containing the compound soluble in water and / or an organic solvent. The present invention has been completed by finding that it is excellent, has excellent permeability to the skin, and exhibits excellent moisturizing property and wettability to the skin.
Embodiments of the present invention are exemplified below.

(1) A composition comprising 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and 2-O-α-D-glycosyl-L-ascorbic acid.
(2)
6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid is the following general formula (3):

(In the formula, n is an integer of 1 to 8, R represents an alkyl group, and the number of carbon atoms as the −C (= O) R group is 12 to 18.)
6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid represented by
2-O-α-D-glycosyl-L-ascorbic acid is the following general formula (1):

(In the formula, n is an integer from 1 to 8.)
The composition according to (1), which is 2-O-α-D-glycosyl-L-ascorbic acid represented by.
(3) The composition according to (1) or (2), which further contains L-ascorbic acid.
(4) The content of 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid is 5 to 60% by weight, 10 to 50% by weight, or 20 to 40% by weight. The composition according to any one of (1) to (3).
(5) The acyl group of 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid has 3 to 24 carbon atoms, 4 to 20, 8 to 20, 12 to 18 or 14 to 16. The composition according to any one of (1) to (4).
(6) 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid contains 6-acyl-2-O-α-D-glucopyranosyl-L-ascorbic acid (1)-(5). The composition according to any one of.
(7) 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid, 6-acyl-2-O-α-D-martosyl-L-ascorbic acid, 6-acyl-2-O- The composition according to any one of (1) to (6), which comprises one selected from the group consisting of α-D- maltotriosyl -L-ascorbic acid or a mixture thereof.
(8) 2-O-α-D-glycosyl-L-ascorbic acid is 2-O-α-D-maltosyl-L-ascorbic acid, 2-O-α-D-maltotriosyl-L-ascorbic acid. The composition according to any one of (1) to (7), which comprises one selected from the group consisting of or a mixture thereof.
(9) The composition according to (8), wherein 2-O-α-D-glycosyl-L-ascorbic acid comprises 2-O-α-D-glucopyranosyl-L-ascorbic acid .
(10) A powder comprising the composition according to any one of (1) to (9).
(11) An external skin preparation containing the composition according to any one of (1) to (9).
(12) A step of preparing a solution in which the composition according to any one of (1) to (9) is dissolved in a solvent of water and / or alcohol, and by spray-drying the solution, 6-acyl- A composition containing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid, which comprises a step of powdering a composition containing 2-O-α-D-glycosyl-L-ascorbic acid. Manufacturing method of things.
(13) The following general formula (4):

(In the formula, R represents a hydrocarbon group.)
6-Acyl-2-O-α-D-maltosyl-L-ascorbic acid represented by.
(14) The 6-acyl-described in (13), wherein R in the general formula (4) is an alkyl group and the number of carbon atoms as the —C (= O) R group is 12 to 18. 2-O-α-D-maltosyl-L-ascorbic acid.
(15) The powder comprising 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid according to (13) or (14).
(16) A skin external preparation containing 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid according to (13) or (14).
(17) A step of preparing a solution in which the 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid according to (13) or (14) is dissolved in a solvent of water and / or alcohol. 6-Acyl-2-O-α-D-Maltsyl-, which comprises a step of pulverizing 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid by spray-drying the solution. A method for producing a powder of L-ascorbic acid.

In the first embodiment of the invention, 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid has excellent solubility in alcohol and water, excellent permeability to the skin, and / or excellent. Shows moisturizing properties and wettability to the skin. 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid has L-ascorbic acid-like bioactivity, has excellent permeability to the skin, and has an epidermis deeper than the stratum corneum of the skin. And / or because it can penetrate into the dermis, it is useful as a skin whitening agent.
In the second embodiment of the present invention, the composition containing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and 2-O-α-D-glycosyl-L-ascorbic acid is water. And / or show excellent solubility in organic solvents, excellent permeability to the skin, and / or excellent moisturizing and wettability to the skin.
In one embodiment of the present invention, 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid is a raw material for external skin preparations such as cosmetics.

The present invention includes a plurality of embodiments. Terms used and / or defined with respect to one embodiment are considered to have the same meaning with respect to all other embodiments, unless otherwise specified.

<2-O-α-D-glycosyl-L-ascorbic acid>
In one embodiment of the present invention, 2-O-α-D-glycosyl-L-ascorbic acid is the following general formula (1).

(In the formula, n indicates an integer of 1 or more.)
It is one kind of 2-O-α-D-glycosyl-L-ascorbic acid represented by or a mixture thereof. N in the general formula (1) is an integer of 1 or more meaning the number of sugars (glucose) bound to the α-position of the glucose residue bound to the 2-position of L-ascorbic acid, and 1 to 8 Alternatively, 1 to 3 is preferable, and 1 is particularly preferable. The 2-O-α-D-glycosyl-L-ascorbic acid of the present invention is 2-O-α-D-glucopyranocil-L-ascorbic acid (n = 1), 2-O-α-D-maltosyl-L. -Ascorbic acid (n = 2), 2-O-α-D-maltotriosyl-L-ascorbic acid (n = 3), 2-O-α-D-maltotetraosyl-L-ascorbic acid (n) = 4), 2-O-α-D-maltopentaosyl-L-ascorbic acid (n = 5), 2-O-α-D-maltohexaosyl-L-ascorbic acid (n = 6), Either 2-O-α-D-maltoheptaosyl-L-ascorbic acid (n = 7) or 2-O-α-D-maltooctaosyl-L-ascorbic acid (n = 8) or It may be a mixture thereof. 2-O-α-D-glycosyl-L-ascorbic acid is at least 2-O-α-D-glucopyranocil-L-ascorbic acid, 2-O-α-D-maltosyl-L-ascorbic acid, and / Alternatively, it preferably contains 2-O-α-D-maltotriosyl-L-ascorbic acid.
As used herein, 2-O-α-D-glycosyl-L-ascorbic acid having a specific glucose number contains other 2-O-α-D-glycosyl-L-ascorbic acid as impurities. The purity may be 90 mol% or more, 95 mol% or more, 98 mol% or more, 99 mol% or more, 99.5 mol% or more, and 99.9 mol% or more. In particular, 2-O-α-D-glycosyl-L-ascorbic acid having n = 4 or more as an impurity is 10 mol% or less, 5 mol% or less, 2 mol% or less, 1 mol% or less, 0.5 mol. %, 0.1 mol% or less, and preferably not substantially contained.

2-O-α-D-glycosyl-L-ascorbic acid can be produced by a known method, for example, the method described in Patent Document 2. As an example, a method for producing 2-O-α-D-glucopyranosyl-L-ascorbic acid is described below.
(1) Containing 2-O-α-D-glucopyranocil-L-ascorbic acid by allowing glycosyltransferase to act on a solution containing L-ascorbic acid and an α-glucosyl sugar compound and then glucoamylase. Make a solution.
(2) A high-purity 2-O-α-D-glucopyranosyl-L-ascorbic acid-containing solution obtained by purifying a 2-O-α-D-glucopyranosyl-L-ascorbic acid-containing solution using an ion exchange resin or the like. To manufacture.
(3) 2-O-α-D-glucopyranosyl-L-ascorbic acid is crystallized to obtain 2-O-α-D-glucopyranosyl-L-ascorbic acid.

Further, 2-O-α-D-martosyl-L-ascorbic acid (n = 2), which substantially does not contain 2-O-α-D-glycosyl-L-ascorbic acid having n = 4 or more, is, for example, as follows. It can be obtained by a process such as.
(1) Glycosyltransferase is allowed to act on a solution containing L-ascorbic acid and an α-glucopyranosyl sugar compound, and 2-O-α-D-glycosyl-L-ascorbic acid (n = 1 to 7) is mixed. A solution is obtained, and then β-amylase is allowed to act on the mixed solution to cleave the sugar chain having n = 4 or more, and 2-O-α-D-glycosyl-L-ascorbic acid (n = 1-3). Make a containing solution.
(2) Purify the 2-O-α-D-glycosyl-L-ascorbic acid (n = 1-3) -containing solution by removing unreacted L-ascorbic acid and saccharides using an ion exchange resin or the like. do.
(3) The solvent is removed to obtain powdered 2-O-α-D-glycosyl-L-ascorbic acid (n = 1 to 3).
The content ratio of 2-O-α-D-maltosyl-L-ascorbic acid can be controlled by the purification conditions of the step (2).

<6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid>
In one embodiment of the invention, 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid has an acyl group at the 6-position of 2-O-α-D-glycosyl-L-ascorbic acid. The following general formula (3):

(In the formula, R represents a hydrocarbon group. N is the same as above.)
It is a compound indicated by.
The hydrocarbon group represented by R in the general formula (3) may be a linear, branched or cyclic, saturated or unsaturated hydrocarbon group, preferably an alkyl group, an alkenyl group, or an alkynyl group. Yes, more preferably a straight chain alkyl group. The carbon number of the hydrocarbon group shall be 3 to 24, 4 to 20, 8 to 20, 12 to 18 when counted as the carbon number of the acyl group, that is, the —C (= O) R group. Is preferable, and 14 to 16 is particularly preferable.
Similar to the general formula (1), n in the general formula (3) means the number of sugars (glucose) bound to the α-position of the glucose residue bound to the 2-position of L-ascorbic acid 1. These integers are preferably 1 to 8 or 1 to 3, and 1 is particularly preferable. 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid is 6-acyl-2-O-α-D-glucopyranocil-L-ascorbic acid (n = 1), 6-acyl-2- O-α-D-maltosyl-L-ascorbic acid (n = 2), 6-acyl-2-O-α-D-maltotriosyl-L-ascorbic acid (n = 3), 6-acyl-2- O-α-D-maltotetraosyl-L-ascorbic acid (n = 4), 6-acyl-2-O-α-D-maltopentaosyl-L-ascorbic acid (n = 5), 6- Acyl-2-O-α-D-maltohexaosyl-L-ascorbic acid (n = 6), 6-acyl-2-O-α-D-maltoheptaosyl-L-ascorbic acid (n = 7) ), Or 6-acyl-2-O-α-D-maltooctaosyl-L-ascorbic acid (n = 8) or a mixture thereof. 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid is at least 6-acyl-2-O-α-D-glucopyranocil-L-ascorbic acid, 6-acyl-2-O-α. It preferably contains -D-maltosyl-L-ascorbic acid and / or 6-acyl-2-O-α-D-maltotriosyl-L-ascorbic acid.

In one embodiment of the invention, 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid with a particular glucose number is the other 6-acyl-2-O-α- as an impurity. It may contain D-glycosyl-L-ascorbic acid and / or an unacylated 2-O-α-D-glycosyl-L-ascorbic acid. 6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid is 90 mol% or more, 95 mol% or more, 98 mol% or more, 99 mol% or more, 99.5 mol% or more, 99.9 mol. The purity is preferably% or higher. 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and / or 2-O-α-D-glycosyl-L-ascorbic acid having n = 4 or more as impurities is 10 mol% or less. It is preferably 5 mol% or less, 2 mol% or less, 1 mol% or less, 0.5 mol% or less, 0.1 mol% or less, and it is preferable that it is substantially not contained. 2-O-α-D-glycosyl-L-ascorbic acid, which is an unacylated product, as an impurity is 10 mol% or less, 5 mol% or less, 2 mol% or less, 1 mol% or less, 0.5 mol%. Hereinafter, it is preferably 0.1 mol% or less, and it is preferable that it is substantially not contained.

6-Acyl-2-O-α-D-glycosyl-L-ascorbic acid can be produced by a method of substituting an acyl group at the 6-position of 2-O-α-D-glycosyl-L-ascorbic acid. As a method for substituting the acyl group at the 6-position, a known method, for example, the method described in Patent Document 4 can be used.
As a method for producing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid, 2-O-α-D-glycosyl-L-ascorbic acid is mixed with an organic acid, an acid halide, an acid anhydride, and an acid ester. Examples thereof include a method of chemically reacting with an acylating agent such as. As the acylating agent, a saturated or unsaturated fatty acid having 3 to 24 carbon atoms, 4 to 20, 8 to 20, 12 to 18, preferably 14 to 16 or an anhydride thereof is preferable, and for example, lauric acid. , Myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid and ricinoleic acid, and anhydrides thereof are preferable.

In one embodiment of the invention, 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid, 2-O-α-D-glycosyl-L-ascorbic acid, and L-ascorbic acid are free acids. Or a salt, and the salt may be an inorganic or organic cation salt. Inorganic cations are metal ions of group 1 elements such as sodium and potassium ions; metal ions of group 2 elements such as magnesium and calcium ions; transition metal ions such as iron and copper ions; and It may be a metal ion selected from the group consisting of other metal ions such as zinc ion and aluminum ion, or a combination thereof. By salting 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and 2-O-α-D-glycosyl-L-ascorbic acid, the pH becomes neutral and other substances. It can make it difficult to decompose other substances when mixed with.

<Method for producing powder of 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid>
A step of preparing a solution in which the 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid is dissolved in a solvent; and by spray-drying the solution, 6-acyl-2-O- A powder of 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid can be produced by a method comprising the step of pulverizing α-D-glycosyl-L-ascorbic acid.
As the solvent, a known solvent can be freely used as long as the composition is soluble, but water and / or alcohol is preferable, and water is particularly preferable. Ethanol is preferable as the alcohol.
The spray drying can be performed using, for example, a spray dryer. The spray drying is preferably performed at the inlet temperature of the spray dryer of 140 to 180 ° C.
If the temperature is lower than 140 ° C., the solvent may not be sufficiently evaporated, so that a dry powder may not be obtained. On the other hand, when the temperature is higher than 180 ° C., 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and 2-O-α-D-glycosyl-L-ascorbic acid are decomposed or volatilized during the process. There is a risk of doing it. Further, it is preferable that the outlet temperature of the spray dryer is 50 to 100 ° C.

<Composition containing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid>
6-Acyl-2-O-α-D- by mixing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and 2-O-α-D-glycosyl-L-ascorbic acid It can be a glycosyl-L-ascorbic acid-containing composition. In one embodiment of the invention, the composition may further comprise L-ascorbic acid. Such compositions have excellent effects on solubility in organic solvents and permeation through the skin. The content of 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid in the composition is preferably 5 to 60% by weight, preferably 10 to 50% by weight, and more preferably 20 to 40% by weight. preferable. Within this range, a particularly excellent effect is exhibited in the solubility in an organic solvent and the permeation performance into the skin. Also, a glucosyl group (C 6H ₁₁ ) with respect to the total weight of ₆ -acyl-2-O-α-D-glycosyl-L-ascorbic acid and 2-O-α-D-glycosyl-L-ascorbic acid in the composition. The weight ratio of O ₅ ) is preferably 30% to 60%, more preferably 35% to 55%.

<Method for producing powder of 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid-containing composition>
A step of preparing a solution in which the 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid-containing composition is dissolved in a solvent, and a step of spray-drying the solution to 6-acyl-2- The powder of the 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid-containing composition is powdered by a method comprising, and a method of powdering the O-α-D-glycosyl-L-ascorbic acid-containing composition. Can be manufactured.
As the solvent, a known solvent can be freely used as long as the composition is soluble, but water and / or alcohol is preferable, and water is particularly preferable. Ethanol is preferable as the alcohol.
The spray drying can be performed using, for example, a spray dryer. The spray drying is preferably performed at the inlet temperature of the spray dryer of 140 to 180 ° C.
If the temperature is lower than 140 ° C., the solvent may not be sufficiently evaporated, so that a dry powder may not be obtained. On the other hand, if the temperature is higher than 180 ° C., 2-O-α-D-glycosyl-L-ascorbic acid may be decomposed or volatilized during the process. Further, it is preferable that the outlet temperature of the spray dryer is 50 to 100 ° C.

In one embodiment of the invention, the composition containing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and / or the compound has high solubility in organic solvents and high skin permeability. It has excellent skin compatibility and skin permeability, and is useful as an external skin preparation for cosmetics and the like.
The external skin preparation means a composition applied to the outer skin, and includes, for example, cosmetics, pharmaceuticals, and quasi-drugs. The rind includes hair and nails.
As the external skin preparation, known bases and carriers generally used in the preparation of external preparations for skin can be appropriately selected and applied according to the purpose of use, the characteristics of the preparation and the form of use.
In addition, if necessary, other components such as surfactants, skin absorption promoters, antioxidants, auxiliaries, UV absorbers, thickeners, bulking agents, stabilizers, colorants, flavoring agents, etc. It can be used in combination with preservatives, fungicides, etc.
External skin preparations include ointments, creams, sprays, lotions, powders, gels, sol agents, aerosols, paps, tapes, liniments, emulsions, lotions, beauty essences, facial masks, and undermakes. As a dosage form for up, foundation, etc., it can be appropriately selected and prepared as desired, such as the purpose of use and the form of use.
In one embodiment of the present invention, the skin external preparation is 0.001 to 50% by weight, 0.01 to 20% by weight, preferably 0.1% by weight to 10% by weight based on 6- of the total skin external preparation. Contains an acyl-2-O-α-D-glycosyl-L-ascorbic acid-containing composition. In one embodiment of the present invention, the skin external preparation is 0.001 to 50% by weight, 0.01 to 20% by weight, preferably 0.1% by weight to 10% by weight based on 6- of the total skin external preparation. Includes acyl-2-O-α-D-maltosyl-L-ascorbic acid. Further, in one embodiment of the present invention, the skin external preparation is preferably 0.001 to 50% by weight, preferably 0.01 to 20% by weight, in terms of weight of L-ascorbic acid, based on the entire skin external preparation. Contains 0.1% by weight to 10% by weight of L-ascorbic acid derivative.

The present invention will be described in more detail in the following examples, but the present invention is not limited thereto.
Other terms and concepts in the present invention are based on the meanings of terms commonly used in the art, and the various techniques used to carry out the present invention are those of particular origin. Except for this, those skilled in the art can easily and surely carry out the procedure based on known documents and the like. In addition, various analyzes were performed by applying the methods described in the analytical instruments or reagents used, the instruction manuals for the kits, catalogs, and the like mutatis mutandis.
The contents of the prior art documents, patent gazettes and patent application specifications cited in the present specification shall be referred to as the description contents of the present invention.

(Example 1)
Preparation of composition containing 6-palmitoyle-2-O-α-D-glucopyranocil-L-ascorbic acid

(1) Synthesis of 2-O-α-D-glucopyranosyl-L-ascorbic acid 125 g of liquefied starch and 50 g of L-ascorbic acid were added to 1000 g of water and dissolved to adjust the pH to 5. To this, 20 g of a cyclomaltodextrin glucanotransferase enzyme preparation (Toruzyme 3.0L, manufactured by Novozimes) was added and reacted at 50 ° C. for 48 hours to cause 2-O-α-D-glucopyranocil-L-ascorbic acid. , 2-O-α-maltosyl-L-ascorbic acid, 2-O-α-maltotriosyl-L-ascorbic acid, 2-O-α-maltotetraosyl-L-ascorbic acid, etc. A reaction solution containing α-D-glycosyl-L-ascorbic acid was obtained.
After inactivating the enzyme by heating the obtained reaction solution to stop the reaction, 1.5 g of a glucoamylase enzyme preparation (glucomylase AF6, manufactured by Amano Enzyme Co., Ltd.) was added, and the temperature was 50 ° C. for 16 hours. By reacting, a reaction solution containing 2-O-α-D-glucopyranosyl-L-ascorbic acid was obtained. After the reaction, the enzyme was inactivated by heating to stop the reaction.

(2) Purification of 2-O-α-D-glucopyranosyl-L-ascorbic acid The obtained reaction solution was heated to inactivate the enzyme to stop the reaction, and then activated carbon was added to decolorize and filter. The filtrate is passed through a column packed with an H-type cation exchange resin to remove metal ions, and then passed through a column filled with an OH-type anion exchange resin to pass 2-O-α-D-glucopyranosyl-L. -Ascorbic acid was adsorbed. The anion exchange resin adsorbed with 2-O-α-D-glucopyranosyl-L-ascorbic acid was washed with water to remove unreacted saccharides, and then 0.1N sodium hydroxide was passed through the solution to 2-O. -Α-D-Glucopyranocil-L-ascorbic acid was eluted. Further, this eluate was passed through a column packed with an H-type cation exchange resin to obtain a purified solution of 2-O-α-D-glucopyranosyl-L-ascorbic acid from which metal ions had been removed.
The obtained purified liquid was concentrated under reduced pressure by heating at 50 ° C., concentrated to a solid content concentration of about 60%, and then the aqueous solution was cooled to 10 ° C. to precipitate crystals. The precipitate was recovered by filtration, washed with a small amount of water, and then dried under reduced pressure to obtain 21 g of 2-O-α-D-glucopyranosyl-L-ascorbic acid powder.

(3) Synthesis of 6-palmityl-2-O-α-D-glucopyranosyl-L-ascorbic acid 20 g of 2-O-α-D-glucopyranosyl-L-ascorbic acid powder was dissolved in 300 ml of anhydrous pyridine at 25 ° C. After adding 20 g of palmitic acid anhydride over 1 hour, the reaction was further carried out at 25 ° C. for 1 hour. Next, the above reaction solution was added to 1500 ml of water, and the resulting precipitate was collected by filtration, washed with 1500 ml of water, and then dried under reduced pressure at 40 ° C.
The obtained reaction product was supported on silica gel chromatography (silica gel 60, manufactured by Nacalai Tesque Co., Ltd.), and then separated and purified using ethyl acetate: methanol = 8: 2 as a developing solvent. The obtained purified liquid was dried under reduced pressure to remove the solvent, then dissolved in ethanol and recrystallized, and the obtained crystals were dried to obtain 6-palmitoyl-2-O-α-D-glucopyranosyl-. 12 g of L-ascorbic acid was obtained.

(4) Spray drying of a composition containing 6-palmitoyle-2-O-α-D-glucopyranosyl-L-ascorbic acid 6-palmitoyle-2-O-α-D-glucopyranosyl-L-ascorbic acid 10 g. A solution dissolved in 100 g of ethanol and an aqueous solution of 10 g of 2-O-α-D-glucopyranosyl-L-ascorbic acid and 10 g of L-ascorbic acid dissolved in 100 g of water are mixed, and the mixed solution is spray-dried (Japan). A composition containing 6-palmitoyle-2-O-α-D-glucopyranosyl-L-ascorbic acid by spray drying at an inlet temperature of 160 ° C. and an outlet temperature of 75 ° C. at B-290) manufactured by Büch. 24 g of powder of the substance was obtained.

(Examples 2 to 6)
The 6-palmitoyle-2-O-α-D-glucopyranosyl-L-ascorbic acid, 2-O-α-D-glucopyranosyl-L-ascorbic acid, and L-ascorbic acid according to (4) of Example 1 are described below. A powder of a composition containing 6-palmitoyle-2-O-α-D-glucopyranocil-L-ascorbic acid was obtained in the same manner as in Example 1 except that the ratio was changed to that shown in 1.
The weight ratio of 6-palmitoyle-2-O-α-D-glucopyranosyl-L-ascorbic acid: 2-O-α-D-glucopyranosyl-L-ascorbic acid: L-ascorbic acid is as follows in each example. did.
(Example 2) 6:47:47
(Example 3) 10:45:45
(Example 4) 20:40:40
(Example 5) 50:25:25
(Example 6) 60:20:20

(Example 7)
6-Lauroyl-2-O-α-D-glucopyranosyl-L-ascorbic acid was contained in the same manner as in Example 1 except that the anhydrous palmitic acid of Example 1 (3) was replaced with anhydrous lauric acid. 18 g of powder of the composition was obtained.

(Example 8)
6-Myristoyl-2-O-α-D-glucopyranosyl-L-ascorbic acid was contained in the same manner as in Example 1 except that the anhydrous palmitic acid of Example 1 (3) was replaced with anhydrous myristic acid. 22 g of powder of the composition was obtained.

(Example 9)
6-Stearoyl-2-O-α-D-glucopyranosyl-L-ascorbic acid was contained in the same manner as in Example 1 except that the anhydrous palmitic acid of Example 1 (3) was replaced with anhydrous stearic acid. 26 g of powder of the composition was obtained.

(Comparative Example 1)
An aqueous solution prepared by dissolving 10 g of 2-O-α-D-glucopyranosyl-L-ascorbic acid and 10 g of L-ascorbic acid in 100 g of water was spray-dried (B-290, manufactured by Nippon Buch) at an inlet temperature of 160 ° C. and an outlet. By spray-drying at a temperature of 75 ° C., 14 g of a powder of the composition was obtained.

(Comparative Example 2)
A solution in which 10 g of sodium palmitate is dissolved in 100 g of ethanol and an aqueous solution in which 10 g of 2-O-α-D-glucopyranosyl-L-ascorbic acid and 10 g of L-ascorbic acid are dissolved in 100 g of water are mixed and mixed. Was spray-dried with a spray dryer (B-290, manufactured by Nippon Buch) at an inlet temperature of 160 ° C. and an outlet temperature of 75 ° C. to obtain 21 g of a powder of the composition.

(Comparative Example 3)
A solution in which 10 g of sodium stearate is dissolved in 100 g of ethanol and an aqueous solution in which 10 g of 2-O-α-D-glucopyranosyl-L-ascorbic acid and 10 g of L-ascorbic acid are dissolved in 100 g of water are mixed and mixed. Was spray-dried with a spray dryer (B-290, manufactured by Nippon Buch) at an inlet temperature of 160 ° C. and an outlet temperature of 75 ° C. to obtain 21 g of a powder of the composition.

<Evaluation of solubility>
Solubility in ethanol 10 g of each composition is added to 20 ml of ethanol, ultrasonic dispersion treatment is performed for 10 minutes, and the solution filtered through a 0.2 μm filter is subjected to high performance liquid chromatography (HPLC) with L-ascorbic acid, 2-. O-α-D-glucopyranosyl-L-ascorbic acid was quantitatively analyzed, and the solubility was calculated and evaluated. The results are shown in Table 1. The dissolution rate is expressed as the ratio of the amount of substance in the solution quantified by HPLC to the amount of substance contained in 10 g of the composition. The HPLC method used a high performance liquid chromatograph device manufactured by Waters, and the column used was Shodex Asahipak NH2P-50 4E 4.6 × 250 mm. (Column temperature: 40 ° C., mobile phase: acetonitrile / 0.1% phosphate buffer = 60/40, flow velocity: 0.7 mL / min, wavelength: 254 nm)

<Evaluation of skin penetration performance>
To 1 g of each composition, 5 ml of ethanol and 5 ml of water were added and ultrasonically dispersed for 10 minutes to dissolve them. 5 ml of the solution was applied to an area of 2 cm × 2 cm of the skin of the forearm and dried.
After 60 minutes had passed after the application, the keratin at the applied site was collected by the tape stripping method. As a stripping tape, OPP tape (375SN, manufactured by 3M Co., Ltd.) was used, and the skin at the same spot to which it was applied was collected three times. The third tape with the skin attached was placed in a closed container, 1 ml of water and 1 ml of ethanol were added, and the mixture was shaken for 1 hour to extract each component. The concentration of L-ascorbic acid and 2-O-α-D-glucopyranosyl-L-ascorbic acid was measured by HPLC in the extracted liquid. The results are shown in Table 2.

The results in Table 1 show that 6-acyl-2-O-α-D-glucosyl-L-ascorbic acid dissolves 2-O-α-D-glucosyl-L-ascorbic acid and L-ascorbic acid in alcohol. It has been shown to have the effect of helping. Even a small amount as in Example 2 has a great effect. Moreover, as is clear from the comparison with Comparative Examples 2 and 3, the effect is much larger than the effect expected from the usual surfactant. The results in Table 2 show that 6-acyl-2-O-α-D-glucosyl-L-ascorbic acid penetrates the skin of 2-O-α-D-glucosyl-L-ascorbic acid and L-ascorbic acid. It has been shown to have the effect of helping.

(Example 10)
Synthesis of 6-palmitoyle-2-O-α-D-maltosyl-L-ascorbic acid (1) Synthesis of 2-O-α-D-glucosyl-L-ascorbic acid (n = 1-3) Liquefaction in 1000 g of water 125 g of starch and 50 g of L-ascorbic acid were added and dissolved to adjust the pH to 5. To this, 20 g of a cyclomaltodexstring lucanotransferase enzyme preparation (Toruzyme 3.0L, manufactured by Novozimes) was added and reacted at 50 ° C. for 48 hours to cause 2-O-α-D-glucopyranocil-L-ascorbic acid. , 2-O-α-D-maltosyl-L-ascorbic acid, 2-O-α-D-maltotriosyl-L-ascorbic acid, 2-O-α-D-maltotetraosyl-L-ascorbic acid A reaction solution containing 2-O-α-D-glycosyl-L-ascorbic acid such as 2-O-α-D-glycosyl-L-ascorbic acid was obtained.
After inactivating the enzyme by heating the obtained reaction solution to stop the reaction, 0.7 g of β-amylase enzyme preparation (β-amylase F "Amano", manufactured by Amano Enzyme Co., Ltd.) was added. By reacting at 50 ° C. for 2 hours, 2-O-α-D-glucopyranosyl-L-ascorbic acid, 2-O-α-D-martosyl-L-ascorbic acid, 2-O-α-D-malt A reaction solution containing triosyl-L-ascorbic acid was obtained. After the reaction, the enzyme was inactivated by heating to stop the reaction.
(2) Purification of 2-O-α-D-maltosyl-L-ascorbic acid Activated carbon was added to the obtained reaction solution for decolorization and filtration. The filtrate is passed through a column packed with an H-type cation exchange resin to remove metal ions, and then passed through a column filled with an OH-type anion exchange resin to pass 2-O-α-D-glucopyranosyl-L. -Ascorbic acid, 2-O-α-D-maltosyl-L-ascorbic acid, 2-O-α-D-maltotriosyl-L-ascorbic acid were adsorbed. The adsorbed anion exchange resin was washed with water to remove unreacted saccharides, and then 0.1N sodium hydroxide was passed through to pass 2-O-α-D-glucopyranosyl-L-ascorbic acid, 2-O. -Α-D-maltosyl-L-ascorbic acid and 2-O-α-D-maltotriosyl-L-ascorbic acid were separated and eluted.
Further, a purified liquid containing 2-O-α-D-maltosyl-L-ascorbic acid was passed through a column packed with an H-type cation exchange resin to remove metal ions. 2-O-α- A purified liquid containing D-maltosyl-L-ascorbic acid was obtained.
(3) Spray drying of 2-O-α-D-maltosyl-L-ascorbic acid An aqueous solution containing 2-O-α-D-maltosyl-L-ascorbic acid is heated and concentrated under reduced pressure to a solid content of about 15%. , Activated carbon was added, decolorization and purification were performed, and the mixture was filtered. The filtrate is spray-dried with a spray dryer (B-290, manufactured by Nippon Buch) at an inlet temperature of 160 ° C and an outlet temperature of 75 ° C to form a powder of 2-O-α-D-maltosyl-L-ascorbic acid. Got
(4) Synthesis of 6-palmitoyle-2-O-α-D-maltosyl-L-ascorbic acid 20 g of 2-O-α-D-maltosyl-L-ascorbic acid powder was dissolved in 300 ml of anhydrous pyridine at 25 ° C. After adding 20 g of palmitic acid anhydride over 1 hour, the reaction was further carried out at 25 ° C. for 1 hour. Next, the above reaction solution was added to 1500 ml of water, and the resulting precipitate was collected by filtration, washed with 1500 ml of water, and then dried under reduced pressure at 40 ° C. The obtained reaction product was supported on silica gel chromatography (silica gel 60, manufactured by Nacalai Tesque Co., Ltd.), and then separated and purified using ethyl acetate: methanol = 8: 2 as a developing solvent. The obtained purified liquid is dried under reduced pressure to remove the solvent, then dissolved in an aqueous ethanol solution, and spray-dried with a spray dryer (B-290, manufactured by Nippon Buch) at an inlet temperature of 170 ° C. and an outlet temperature of 85 ° C. By doing so, 14 g of a powder of 6-palmitoyle-2-O-α-D-maltosyl-L-ascorbic acid was obtained.

Identification of 6-palmitoyle-2-O-α-D-maltosyl-L-ascorbic acid
¹ H NMR (300 MHz, CD ₃ OD): δ 0.90 (t, J = 6.9 Hz, 3H), 1.28 (m, 24H), 1.62 (qn, J = 6.9 Hz, 2H), 2.35 (t, J = 6.9 Hz, 2H), 3.26 (dd, J = 10.3, 9.3 Hz, 1H), 3.45 (dd, J = 9.6, 3.9 Hz, 1H), 3.58 (dd, J = 9.9, 3.6 Hz, 1H), 3.60 ( dd, J = 10.0, 9.6 Hz, 1H), 3.65 (dd, J = 12.0, 2.4 Hz, 1H), 3.66 (dd, J = 12.6, 3.6 Hz, 1H), 3.67 (dd, J = 9.9, 9.6 Hz) , 1H), 3.77 (dd, J = 12.6, 2.2 Hz, 1H), 3.78 (ddd, J = 10.0, 3.6, 2.2 Hz, 1H), 3.84 (dd, J = 12.0, 3.6 Hz, 1H), 4.05 ( dd, J = 9.6, 9.3 Hz, 1H), 4.09 (ddd, J = 10.3, 3.6, 2.4 Hz, 1H), 4.11 (ddd, J = 7.2, 6.0, 1.5 Hz, 1H), 4.15 (dd, J = 10.8, 6.0 Hz, 1H), 4.27 (dd, J = 10.8, 7.2 Hz, 1H), 4.80 (d, J = 1.5 Hz, 1H), 5.20 (d, J = 3.6 Hz, 1H), 5.42 (d, J = 3.9 Hz, 1H).
¹³ C NMR (75 MHz, CD ₃ OD): δ 14.4, 23.7, 26.0, 30.2, 30.4, 30.5, 30.6, 30.8 (6), 33.1, 34.9, 61.5, 62.7, 65.6, 67.9, 71.5, 73.1, 73.3, 74.2, 74.3, 74.8, 75.1, 77.5, 80.8, 101.1, 102.9, 119.8, 163.2, 172.3, 175.1.
MS: ESI m / z [MH] ^- : Calculated value (C ₃₄ H ₅₈ O ₁₇ ) 737.3595; Measured value 737.3624.
MS was measured using an Insertsil Ph analysis column (φ4.6 × 250 nm, GL Sciences INC.). Measurement conditions are 80% MeOH aqueous solution of 1% acetic acid; 0.3 ml / min; 40 ° C; UV 240 nm. Measurement mode: ESI (ES-) capillary voltage; 3.5 V, cone voltage; 150 V, source temperature; 120 ° C. there were.

(Comparative Example 4)
Synthesis of 6-palmityl-2-O-α-D-glucopyranosyl-L-ascorbic acid 20 g of powder of 2-O-α-D-glucopyranosyl-L-ascorbic acid was dissolved in 300 ml of anhydrous pyridine and anhydrous at 25 ° C. After adding 29 g of palmitic acid over 1 hour, the reaction was further carried out at 25 ° C. for 1 hour. Next, the above reaction solution was added to 1500 ml of water, and the resulting precipitate was collected by filtration, washed with 1500 ml of water, and then dried under reduced pressure at 40 ° C. The obtained reaction product was supported on silica gel chromatography (silica gel 60, manufactured by Nacalai Tesque Co., Ltd.), and then separated and purified using ethyl acetate: methanol = 8: 2 as a developing solvent. The obtained purified liquid is dried under reduced pressure to remove the solvent, then dissolved in an aqueous ethanol solution, and spray-dried with a spray dryer (B-290, manufactured by Nippon Buch) at an inlet temperature of 170 ° C. and an outlet temperature of 85 ° C. By doing so, 12 g of a powder of 6-palmitoyle-2-O-α-D-glucopyranosyl-L-ascorbic acid was obtained.

(Comparative Example 5)
Synthesis of 6-palmitoyle-2-O-α-D-maltotriosyl-L-ascorbic acid 20 g of powder of 2-O-α-D-maltotriosyl-L-ascorbic acid was dissolved in 300 ml of anhydrous pyridine and 25 After adding 15 g of palmitic acid anhydride at ° C for 1 hour, the reaction was further carried out at 25 ° C for 1 hour. Next, the above reaction solution was added to 1500 ml of water, and the resulting precipitate was collected by filtration, washed with 1500 ml of water, and then dried under reduced pressure at 40 ° C. The obtained reaction product was supported on silica gel chromatography (silica gel 60, manufactured by Nacalai Tesque Co., Ltd.), and then separated and purified using ethyl acetate: methanol = 8: 2 as a developing solvent. The obtained purified liquid is dried under reduced pressure to remove the solvent, then dissolved in an aqueous ethanol solution, and spray-dried with a spray dryer (B-290, manufactured by Nippon Buch) at an inlet temperature of 170 ° C. and an outlet temperature of 85 ° C. By doing so, 10 g of a powder of 6-palmitoyle-2-O-α-D-maltotriosyl-L-ascorbic acid was obtained.

(Experimental example)
<Evaluation of solubility in water and ethanol>
10 g of each ascorbic acid derivative synthesized in Examples 10 and Comparative Examples 4 and 5 was added to 20 ml of water or ethanol, subjected to ultrasonic dispersion treatment for 10 minutes, and the solution filtered through a 0.2 μm filter was subjected to high performance liquid chromatography. Each ascorbic acid derivative was quantitatively analyzed by the method (HPLC), and the solubility was calculated and evaluated. The results are shown in Table 3 . The dissolution rate is expressed as the ratio of the amount of substance in the solution quantified by HPLC to the amount of substance contained in 10 g of each ascorbic acid derivative. The HPLC method used a high performance liquid chromatograph device manufactured by Waters, and the column used was Inertsil Ph-3 HP 3 μm 4.6 × 250 mm (column temperature: 40 ° C., mobile phase: methanol / 1% acetic acid aqueous solution = 80/20). , Flow velocity: 0.6 mL / min, wavelength 240 nm). The results are shown in Table 3.

* The values in the table indicate the dissolution rate (%).

<Evaluation of skin penetration performance>
To 0.1 g of each ascorbic acid derivative synthesized in Examples 10 and Comparative Examples 4 and 5, 4 ml of ethanol and 6 ml of water were added to perform ultrasonic dispersion treatment for 10 minutes, and the pH was adjusted to 5 with a dilute aqueous sodium hydroxide solution. 5 ml of the solution was applied to an area of 2 cm × 2 cm of the skin of the forearm and dried. After 7 hours had passed after the application, the keratin at the applied site was collected by the tape stripping method. As a stripping tape, OPP tape (375SN, manufactured by 3M Co., Ltd.) was used, and the skin at the same spot to which it was applied was collected 10 times. The 3rd, 5th, and 10th skin-attached tapes were placed in a closed container, 2 ml of a mixed solvent of water / ethanol = 4: 6 was added, and the mixture was shaken for 1 hour to extract each component. The concentration of each ascorbic acid derivative was measured by HPLC in the extracted liquid. The results are shown in Table 4.

* The values in the table indicate the concentration (mg / ml).

<Evaluation of moisturizing property>
After applying 10 μL of each sample solution per 1 cm ² to the flexion side of the human forearm in a constant temperature and humidity chamber (25 ° C., 50% RH), the sample solution was left for 30 minutes.
Then, the epidermis conductance (k2) immediately after application was measured with the skin surface stratum corneum water content measuring device [SKION-200 (manufactured by IBS Co., Ltd.)], and the epidermis conductance (k1) immediately before application was measured. The ratio (k2 / k1) of was obtained.
Similarly, for the other eight panelists, this ratio was calculated, and the arithmetic mean value (rounded off to the first decimal place) of the nine panelists was evaluated according to the following evaluation criteria. The larger the value, the higher the moisturizing effect on the skin. The results are shown in Table 5.
(Evaluation criteria)
◎: 1.6 or more ○: 1.3 to 1.5
Δ: 1.0 to 1.2
×: 0.9 or less

<Evaluation of stickiness>
The stickiness was evaluated by 9 panelists.
After applying 0.2 mL of each 1% by mass ethanol / water (4: 6) solution to the forearm, it was judged by sensory evaluation. The evaluation criteria based on the sticky feeling score are as follows.
2 points: preferred 1 point: slightly preferred 0 points: usually -1 point: slightly unfavorable -2 points: unfavorable Next, calculate the arithmetic mean value of 9 panelists (rounded to the first decimal place) and calculate the following. The evaluation criteria were evaluated on a scale of 3 from ○ to ×. The results are shown in Table 3.
○: 1.0 points or more (does not feel sticky)
Δ: -0.5 to 0.9 (slightly sticky feeling)
×: -0.6 points or less (feels a lot of stickiness)

As a result of the experiment, 6-palmitoyl-2-O-α-D-maltosyl-L-ascorbic acid of Example 10 has excellent solubility in alcohol and water, and has high permeability to the skin. all right. Compared with the compounds of Comparative Example 4 and Comparative Example 5, the disaccharide 6-palmitoyle-2-O-α-D-maltosyl-L-ascorbic acid is surprisingly related to the order of sugar chain length. However, it specifically showed excellent solubility and permeability to the skin. Furthermore, in the evaluation of moisturizing property and stickiness, the properties equal to or better than those of the comparative example were shown in both respects.

The composition containing 6-acyl-2-O-α-D-glycosyl-L-ascorbic acid and / or the compound of the present invention is particularly useful as a skin external preparation such as cosmetics.

Claims (5)

The following general formula (4):

(In the formula, R indicates a hydrocarbon group.)
6-Acyl-2-O-α-D-maltosyl-L-ascorbic acid represented by. The 6-acyl-2-O according to claim 1, wherein R in the general formula (4) is an alkyl group and the number of carbon atoms as the —C (= O) R group is 12 to 18. -Α-D-maltosyl-L-ascorbic acid. The powder comprising 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid according to claim 1 or 2. A skin external preparation containing 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid according to claim 1 or 2. A step of preparing a solution in which 6-acyl-2-O-α-D-maltosyl-L-ascorbic acid according to claim 1 or 2 is dissolved in a solvent of water and / or alcohol, and a step of spray-drying the solution. By How to make powder.