JP5244400B2 - Alkylresorcinol glycoside, its production method and use - Google Patents

Description

  The present invention relates to a novel 4-alkylresorcinol glycoside, its production method, and use, and more specifically, 4-alkylresorcinol or a derivative thereof (excluding glycosides, hereinafter referred to as 4-alkylresorcinol in the present specification). And its derivatives (excluding glycosides may be referred to as “4-alkylresorcinols” in some cases)), and 4-alkylresorcinol glycosides in which one or two glycosyl groups are bonded, and The present invention relates to a production method and a composition containing the 4-alkylresorcinol glycoside.

  4-Alkylresorcinols are effective for prevention and improvement of skin pigmentation (Japanese Patent Laid-Open No. 2001-206913), whitening (Japanese Patent Laid-Open No. 2002-179516), suppression of pruritus (Japanese Patent Laid-Open No. 2001-302506), etc. It is also known that it can be used as a component of an external composition for skin, and has an antibacterial action against bacteria that are involved in the generation and deterioration of acne, so that it can be used as a component of an acne-suppressing cosmetic (Japanese Patent No. 2875374). Therefore, 4-n-butylresorcinol is already used as an active ingredient in quasi-drugs for external preparations for skin.

  However, 4-alkylresorcinols generally have a number of problems such as low solubility in water and difficulty in handling, poor stability after dissolution, and irritation. Therefore, it is inconvenient when used as a quasi-drug, cosmetics, etc., and development of a method for using these in a stabilized, water-soluble and low-stimulated derivative is desired.

  In addition, attempts have been made to convert the alkyl group at the 4-position for the purpose of improving the transdermal absorbability of alkylresorcinol, and 4-cyclopentylresorcinol, 4-cyclohexylresorcinol (for example, JP 2005-527534 A, JP 2005). And a branched alkyl group such as 4- (1-methylpropyl) resorcinol, 4- (1-methylbutyl) resorcinol, etc. having a cyclic alkyl group such as JP-A-526093 and JP-A-11-152203) Resorcinol derivatives have been developed (see, for example, JP 2006-124358 A and JP 2006-124357 A). Although these compounds are less effective in inhibiting melanin production, they have improved transdermal absorbability and were expected to be useful active ingredients, but their stability is impaired compared to that of linear alkyl groups, Development of a technique for ensuring this stability has been desired.

  Furthermore, in order to improve the water solubility of resorcinol, Japanese Patent Publication No. 7-36759 and Japanese Patent Application Laid-Open No. 2001-46096 propose a method for producing a glycoside using a glycosyltransferase. No. 2002-179516 describes a whitening composition containing an alkylresorcinol derivative. However, no effective means for solving the drawbacks that have been an obstacle to the use of these alkylresorcinols has been developed yet, and the above-mentioned patent documents all describe the alkylresorcinol glycosides. unacceptable.

  The production of 4-alkylresorcinol, which is a raw material for producing a glycoside of 4-alkylresorcinol and / or its acylated product, is already known (for example, JP-A-2-49715, Lille Bitter, L. A .; see Peiner, V. Trudy-Nauchono-Issledovatel 'skii Institute Sltsev (1969), No. 18, 127-34). As for the method of glycosylation of a phenolic substance, for example, CGTase (cyclodextrin glucanotransferase) [EC 2.4.1.19], α-amylase [EC 3.2.1.1], β-amylase [ EC 3.2.1.2], glucoamylase [EC 3.2.1.3], α-glucosidase [EC 3.2.1.20], β-galactosidase [EC 3.2.1.23] Glucose transfer reactions utilizing bacterial enzymes such as those described above are known (see, for example, JP-A Nos. 2004-115392 and 2001-238673).

  The present invention eliminates the drawbacks of alkylresorcinols, is excellent in water solubility and stability, is substantially tasteless, odorless, does not irritate, has no concern for toxicity, and has sufficient physiological activity in vivo. It is an object to provide an alkylresorcinol derivative that can be exerted.

  As a result of intensive studies to solve the above-mentioned problems, the present inventor has found that glycosides of alkylresorcinols in which one or two glycosyl groups are α-linked to alkylresorcinols (hereinafter collectively referred to as “ Alkylresorcinol glycosides ") may be easily hydrolyzed in vivo, with excellent water solubility, stability, virtually tasteless, odorless, less irritating, no toxic concerns, The present inventors have found that alkylresorcinols exhibit their original physiological activity. Also, a method for producing alkylresorcinol glycosides in large quantities at low cost by an organic synthesis method or an enzyme method, particularly an enzyme method using glycosyltransferase, was found, and this alkylresorcinol glycoside was further incorporated. And the present invention was completed.

  The alkylresorcinol glycoside of the present invention is more stable than alkylresorcinols, exhibits high water solubility, is odorless, less irritating, has no toxicity concerns, and is easily hydrolyzed in vivo. And exhibits the physiological activity inherent in alkylresorcinols. In addition, the alkylresorcinol glycoside of the present invention can be blended at a high concentration in compositions such as pharmaceuticals, quasi-drugs, and cosmetics, and is effective in the whitening effect of alkylresorcinols and the therapeutic effect on anti-sensitive diseases. An excellent composition can be produced. Furthermore, according to the method for producing an alkylresorcinol glycoside of the present invention, an alkylresorcinol glycoside can be produced in a large amount at a low cost from alkylresorcinols and, in particular, an inexpensive partially decomposed starch.

It is a figure which shows the UV absorption spectrum of 4-n-butyl resorcinol glycoside of this invention. It is a figure which shows the elution pattern of HPLC (CAPCELLLPAK C18 AG120 column use) of the 4-n-butyl resorcinol glycoside containing preparation of this invention prepared by the glycosyltransferase reaction which uses a glycosyltransferase. MS of the reaction product a (3-O-α-D-glucopyranosyl-4-n-butylresorcinol) and the reaction product b (1-O-α-D-glucopyranosyl-4-n-butylresorcinol) of the present invention It is a figure which shows a spectrum. Is a diagram showing ¹ H-NMR spectrum of the reaction product a (3-O-α- D- glucopyranosyl -4-n-butyl resorcinol) of the present invention. It is a figure which shows the ¹³ C-NMR spectrum of the reaction product a (3-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of this invention. It is a figure which shows the HH COSY spectrum of the reaction product a (3-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of this invention. It is a figure which shows the C—H COSY spectrum of the reaction product a (3-O-α-D-glucopyranosyl-4-n-butylresorcinol) of the present invention. Is a diagram showing ¹ H-NMR spectrum of the reaction product b (1-O-α- D- glucopyranosyl -4-n-butyl resorcinol) of the present invention. It is a figure which shows the ¹³ C-NMR spectrum of the reaction product b (1-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of this invention. It is a figure which shows the HH COSY spectrum of the reaction product b (1-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of this invention. It is a figure which shows the C-H COSY spectrum of the reaction product b (1-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of this invention. It is a figure which shows the remote C-H COSY spectrum of the reaction product b (1-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of this invention. It is the figure which expanded a part of remote CH COSY spectrum of the reaction product b (1-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol) of FIG.

The 4-alkylresorcinol glycoside referred to in the present invention is a 4-alkylresorcinol glycoside in which a glycosyl group is bonded to either R ₁ or R ₂ of the 4-alkylresorcinol represented by the following general formula 1. The glycosyl group may be a 4-alkylresorcinol glycoside bonded to two positions of R ₁ and R ₂ , or a mixture of two or more of these glycosides. Good.

（但し、式中Ｒ_１、Ｒ_２はそれぞれ独立に水素原子またはグリコシル基を表し、且つ、Ｒ_１、Ｒ_２の少なくとも何れかはグリコシル基であり、Ｒ_３は炭素数３乃至２０の環状構造又は分岐構造を有するアルキル基、或いは、炭素数１乃至２０の直鎖構造を有するアルキル基を表す。）

(In the formula, R ₁ and R ₂ each independently represent a hydrogen atom or a glycosyl group, and at least _one of R ₁ and R ₂ is a glycosyl group, and R ₃ is a cyclic structure having 3 to 20 carbon atoms. Alternatively, it represents an alkyl group having a branched structure or an alkyl group having a linear structure having 1 to 20 carbon atoms.)

  The 4-alkylresorcinol glycoside of the present invention may have a cyclic or branched structure in which the alkyl group has 3 to 20 carbon atoms, preferably 4 to 18 alkyl groups, It may have 1 to 20 unsubstituted linear structures. The cyclic structure is not limited to an aliphatic group such as an alkyl group or an alkenyl group. For example, an aromatic group such as a phenyl group is included as long as the portion directly bonded to resorcinol is an alkyl group. Further, when such an aromatic cyclic substituent exists, the carbon number is counted including the carbon number of the aromatic substituent. Examples of the alkyl group having a cyclic or branched structure include a cyclopentyl group, a cyclohexyl group, a 2-phenylethyl group, an isooctyl group, an amyl group, an isoamyl group, an isobutyl group, a tertiary butyl group, a 1-methylpropyl group, 1 -Methylbutyl group, 1-ethylpropyl group, 1-ethyl-2-methylpropyl group, 1-isopropyl-2-methylpropyl group, 1-dimethyl-3-methylbutyl group, 1-butylpentyl group, 1-isobutyl-3 -Methylbutyl group, isopropyl group, isobutyl group, isoamyl group, 2-methylhexyl group or isostearyl group can be preferably exemplified. Of these, preferred are those having a cyclic alkyl group, preferably a cyclopropyl group having low toxicity, and those having a branched alkyl group are preferably a 1-methylpropyl group having a good balance between effect and toxicity. Is preferably a 1-methylbutyl group. In terms of the remarkable effects of the present invention, a 1-ethylpropyl group or 1-isopropyl-2-methylpropyl group is preferable because a significant improvement in stability can be obtained.

  Examples of the alkyl group having an unsubstituted linear structure include a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, Examples include dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl and eicosyl groups. Among these, those having an alkyl group having 1 to 7 carbon atoms are desirable from the viewpoint of melanin production inhibitory activity and anti-inflammatory activity, and butyl group is particularly desirable.

  Furthermore, in the 4-alkylresorcinol glycoside of the present invention, one or more hydrogen atoms of the alkyl group include a substituent containing an oxygen atom such as a hydrocarbon group other than a methyl group, a carboxyl group, or a ketone group. , A substituent containing a nitrogen atom such as an amino group or a nitro group, a substituent containing a sulfur atom such as a sulfone group or a sulfide group, or a substituent substituted with a halogen group such as a fluorine group or a chloro group. Moreover, in these 4-alkyl resorcinol glycosides, when an optical isomer exists, as long as it has the physiological activity of 4-alkyl resorcinols such as antibacterial action and whitening action, Even if it exists, both may be mixed.

  In the 4-alkylresorcinol glycoside of the present invention, the glycosyl group bonded to the two hydroxyl groups of alkylresorcinol can be combined with 4-alkylresorcinol by a synthesis method or an enzymatic method, There is no limitation, for example, glucosyl group, galactosyl group, fructosyl group, arabinosyl group, maltosyl group, isomaltosyl group, cerbiosyl group, raffinosyl group, gentiobiosyl group, cordobibiosyl group, laminaribiosyl group, nigerosyl group, sambubiosyl group, neohesperidosyl group Any one or two selected from a cellotriosyl group, a glycerol group or a gentiotriosyl group can be exemplified. Among these, from the viewpoint of ease of production, a glucosyl group and a malto-oligosyl group including a maltosyl group are desirable, and a glucosyl group is particularly desirable.

  There is no restriction | limiting in the origin and manufacturing method of the 4-alkyl resorcinol glycoside of this invention, What was manufactured combining suitably methods, such as an organic synthesis method, a fermentation method, and an enzyme method, may be sufficient. Specifically, an organic synthesis method in which an acylated sugar is condensed with 4-alkylresorcinol in the presence of a silver peroxide catalyst, and an enzyme method using a glycosyltransferase can be preferably exemplified. Among them, an enzymatic method in which a glycosyltransferase is allowed to act on a system containing 4-alkylresorcinols and an α-glycosyl compound is inexpensive in large quantities from 4-alkylresorcinols and inexpensive starches or partial hydrolysates thereof. Since a 4-alkylresorcinol glycoside can be produced, it is excellent in economic efficiency and can be advantageously used industrially.

  There is no limitation on the origin and production method of 4-alkylresorcinols used in the production of the 4-alkylresorcinol glycosides of the present invention. For example, resorcinol and the corresponding carboxylic acid are condensed in the presence of zinc chloride, and zinc amalgam / A method of reducing with hydrochloric acid, a method of condensing resorcinol and the corresponding alcohol at a high temperature of 200 to 400 ° C. (for example, Lille, J .; Bitter, LA; Peiner, V. Trudy-Nauchono-Isledovatel 'skiii) Institute Slantsev (1969), No. 18, 127-34), a method obtained by reacting a saturated carboxylic acid and resorcinol at a high temperature of 200 ° C. to 400 ° C. using an alumina catalyst (for example, British Patent No. 1). 1,581,4 It can be easily obtained by 8 Pat reference) may be used, such as a commercially available product.

  The production method of 4-alkylresorcinols will be described more specifically. For example, if isooctyl group or isostearyl group is introduced, resorcinol and isooctanoic acid or isostearic acid may be condensed in the presence of zinc chloride. If cyclopentyl alcohol, cyclohexyl alcohol, phenethyl alcohol, 2-ethylhexanol, isoamyl alcohol, amyl alcohol, isobutyl alcohol, and tertiary butyl alcohol are condensed with resorcinol at 200 to 300 ° C., 4-cyclopentyl resorcinol, 4- Cyclohexyl resorcinol, 4-isooctyl resorcinol, 4-amyl resorcinol, 4-isoamyl resorcinol, 4-isobutyl resorcinol, 4-tarsha Over-butyl resorcinol and the like can be obtained. Furthermore, when an alkyl group having a methyl group at the 1-position is introduced, the method described in JP-A-2006-124358 and JP-A-2006-124357 may be followed, and acetophenone is converted from propyl chloride or butyl chloride. When the derived Grignard reagent is reacted, 4- (1-methylpropyl) resorcinol, 4- (1-methylbutyl) resorcinol and 4-isostearylresorcinol are obtained. Similarly, when an ethyl group is introduced at the 1-position, the desired derivative can be obtained by catalytic reduction of 2 ′, 4′-dihydroxypropiophenone as a starting material and the remaining hydroxyl group using palladium as a catalyst. . This method is also described in JP-A-2006-124357. The introduction of another alkyl group having a branch at the 1-position can also be obtained by conducting a Grignard reaction using 4-isobutyrylresorcinol or the like as a starting material and, if necessary, a catalytic reduction reaction using palladium as a catalyst. The 4-alkylresorcinol thus obtained tends to be less stable in storage than linear alkyl-substituted resorcinols such as 4-n-butylresorcinol. An example is shown below.

<Reference example>
The stability of aqueous solutions of resorcinol derivatives in which the 4-position hydrogen atom was substituted with an alkyl group was compared. That is, 0.5% aqueous solutions of various 4-alkylresorcinols were prepared, stored for 1 month at 40 ° C., and the results of quantification of 4-alkylresorcinol by HPLC are shown in Table 1. HPLC conditions were as follows: column: ODS 4.6 mm × 150 mm, column temperature: 40 ° C., detection: absorption at 230 nm in ultraviolet region, mobile phase: 15 mM phosphate buffer solution (pH 6.5, 25% acetonitrile-15 mM tetrabutylammonium bromide) ), Flow rate: 1 ml / min. The quantitative value after storage was divided by the quantitative value immediately after production, and a value obtained by multiplying by 100 was obtained as the residual rate.

  As is clear from Table 1, it has been found that when a branched alkyl group or a cyclic alkyl group is employed as the alkyl group, the stability is lowered.

  The production method of the 4-alkylresorcinol glycoside by the enzymatic method of the present invention will be specifically described. The glycosyltransferase used for the production of the glycoside is a solution containing a 4-alkylresorcinol and a glycosyl compound. There is no particular limitation as long as it produces 4-alkylresorcinol glycosides when allowed to act. From the viewpoint of action and effect, an enzyme that generates a glycoside by α-bonding is desirable.

  Specifically, for example, α-glucosidase is an enzyme derived from animal and plant tissues such as pig liver, buckwheat seed, or mold belonging to the genus Mucor, Penicillium, or Saccharomyces. Enzymes derived from cultures obtained by culturing microorganisms such as yeast belonging to the genus etc. in nutrient media are cyclomaltodextrin glucanotransferase derived from bacterial cultures belonging to the genus Bacillus, Klebsiella etc. As the enzyme, α-amylase can be appropriately selected from known enzymes having transglycosylation activity such as bacteria derived from bacteria belonging to the genus Bacillus or enzymes derived from mold cultures belonging to the genus Aspergillus. These glycosyltransferases do not necessarily need to be purified and used. Usually, the object of the present invention can be achieved with a crude enzyme such as a culture solution, concentrated solution or salted-out product of these microorganisms.

  These enzymes may be used after being purified by various known methods as needed, or may be used by immobilizing them on a carrier. Commercially available glycosyltransferases can also be used. Usually, the amount of enzyme is selected so that the reaction is completed in about 5 to 80 hours from the viewpoint of economy. Moreover, when using an immobilized enzyme, the method of performing an enzyme reaction by a batch type or a continuous type can be selected suitably.

  The glycosyl compound used in the present invention is not particularly limited as long as it becomes a substrate for the enzyme when a 4-alkylresorcinol glycoside is produced using a glycosyltransferase. For example, amylose, dextrin, cyclodextrin, malto-oligo Preferred examples include starch partial hydrolysates such as sugar, and α-glucosyl compounds such as liquefied starch and gelatinized starch.

  In order to facilitate the production of a 4-alkylresorcinol glycoside, a glycosyl compound suitable for each enzyme may be selected depending on the glycosyltransferase used.

  The concentration of the glycosyl compound in the reaction solution during the transglycosylation reaction is not particularly limited, but is usually 0.1% by mass or more (hereinafter referred to as “%” unless otherwise specified). And 0.1% to 70% is desirable, and a solution of 0.1% to 50% is particularly desirable.

  In addition, the 4-alkylresorcinol-containing liquid at the time of reaction desirably contains 4-alkylresorcinol as high as possible. For example, 4-alkylresorcinol is dissolved in suspension or at a high temperature. Alternatively, a solution containing a high concentration of 4-alkylresorcinols in a solution form dissolved at an alkaline pH exceeding 7.0 is suitable. Usually, the concentration of 4-alkylresorcinols is about 0.00. 005% or more is used, and a solution of about 0.01% to 10.0% is particularly desirable.

  In the reaction method of the present invention, it is desirable to allow glycosyltransferase to act in a state where the charged concentration of 4-alkylresorcinol is increased. For example, when 4-alkylresorcinols are reacted in suspension, 4-alkylresorcinol containing about 0.1% to 2.0% suspended 4-alkylresorcinols and an appropriate amount of glycosyl compound is used. The liquid with a high concentration is adjusted to a pH of about 4.0 to 7.0, and is maintained at a high temperature at which glycosyltransferase can act, specifically, at about 20 ° C. to 90 ° C., and the glycosyltransferase is allowed to act on this. As the 4-alkylresorcinols are converted to 4-alkylresorcinol glycosides, the suspended 4-alkylresorcinols gradually dissolve, and at the same time, 4-alkylresorcinol glycosides are easily produced at high concentrations. .

  Further, for example, when alkylresorcinols are glycosylated on the alkali side exceeding pH 7.0, about 0.2% to 5.0% alkylresorcinols are added to water having a pH of about 7.5 to 10.0. A solution containing a high amount of alkylresorcinols, which is obtained by dissolving by heating and dissolving an appropriate amount of an α-glycosyl compound therein, has a pH as high as possible at which glycosyltransferase can act, a high temperature, specifically, a pH of about 7.5 to When the temperature is maintained at 10.0 and a temperature of about 20 ° C. to 90 ° C. and a glycosyltransferase is allowed to act on this, an alkylresorcinol glycoside is easily produced at a high concentration. At this time, the alkyl resorcinols in the alkaline solution are liable to be decomposed. Therefore, in order to prevent this, it is desirable to keep them as light-shielded and anaerobic as possible. If necessary, an antioxidant such as L-ascorbic acid or erythorbic acid may coexist.

  Further, for example, a liquid containing a high amount of alkylresorcinols containing about 0.5% to 10.0% alkylresorcinols and an appropriate amount of an α-glycosyl compound is adjusted to a pH of about 7.5 to 10.0 and a temperature of about 50 ° C. When maintained at 80 to 80 ° C. and a glycosyltransferase is allowed to act thereon, an alkylresorcinol glycoside is easily produced at a high concentration.

  Further, as alkylresorcinols, for example, about 0.1% to 1.0N caustic soda aqueous solution, caustic potash aqueous solution, sodium carbonate aqueous solution, calcium hydroxide water, aqueous ammonia, etc. A solution dissolved at a high concentration of 10.0% is used, and an acidic aqueous solution such as hydrochloric acid or sulfuric acid is added thereto to adjust to a pH at which the enzyme can act, preferably to a pH exceeding pH 7.0. It is very advantageous to add a glycosyl compound and immediately act on a glycosyltransferase, because an alkylresorcinol glycoside can be easily produced at a high concentration.

  At this time, the alkylresorcinol solution dissolved in a high concentration also tends to cause precipitation when the pH is adjusted with an acidic aqueous solution. Therefore, before adjusting the pH, α-glycosyl compound or a small amount of alkylresorcinol glycoside It is also possible to advantageously carry out the transglycosylation reaction while suppressing the precipitation of alkylresorcinols in the presence of a body.

  Further, if necessary, in order to increase the solubility of the alkylresorcinols before the reaction and facilitate the sugar transfer reaction to the alkylresorcinols, an organic solvent that can be dissolved in water with the alkylresorcinol-rich solution, for example, Coexistence of lower alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol and acetonitrile acetone, lower ketones and the like can also be selected as appropriate.

  In addition, the relatively high-molecular alkylresorcinol glycoside produced by the transglycosylation reaction may be used as it is or after purification with a porous synthetic adsorption resin, after which glucoamylase (EC 3.2.1.3) is used. ), Or by partial hydrolysis with an amylase such as β-amylase (EC 3.2.1.2) to reduce the number of α-glucosyl groups of α-glycosylalkylresorcinols. For example, when glucoamylase is allowed to act, a high-molecular substance higher than α-maltosyl-4-alkylresorcinols can be hydrolyzed to produce glucose and accumulatively produce α-glucosyl-4-alkylresorcinols. In addition, when β-amylase is allowed to act, a polymer higher than α-maltotriosyl-4-alkylresorcinol is hydrolyzed to produce maltose and mainly α-glucosyl-4-alkyl. A mixture of resorcinols and α-maltosyl-4-alkylresorcinols can be accumulated.

  The 4-alkylresorcinol glycoside that can be obtained by the method for producing a glycoside by the enzymatic method of the present invention is different from the hydroxyl group of 4-alkylresorcinol in the bonding mode of the glucosyl group to the hydroxyl group of 4-alkylresorcinol. Three types are obtained: one in which a glycosyl group is introduced and one in which a glycosyl group is introduced at both the 1-position and the 3-position. In this case, the number of monosaccharides constituting each glycosyl group is not particularly limited, but from the viewpoint of ease of production and ease of handling, usually 1 to 6 monosaccharides are desirable. And / or two are desirable, and one is particularly desirable. The reaction solution obtained by the production method of the present invention is partially purified as it is, and then highly purified. Further, these three types of glycosides are individually fractionated and fractionated. It is also optional to mix and mix these with a syrup filtered and concentrated by a conventional method, or after drying and pulverizing the composition.

  The purification method of these 4-alkylresorcinol glycosides is not particularly limited as long as it can increase the purity of the 4-alkylresorcinol glycosides. Usually, the adsorptivity by a porous synthetic adsorbent is used. Using the difference, a 4-alkylresorcinol glycoside and a contaminant such as an α-glycosyl compound may be separated and purified. In this case, non-glycosylated 4-alkylresorcinols themselves have physiological activity and need not be removed. However, from the viewpoint of improving water solubility, 4-alkylresorcinol glycosides are possible as much as possible. It is desirable to increase the purity.

  The porous synthetic adsorbent referred to in the present invention is a synthetic resin such as a porous, wide adsorption surface area, and nonionic styrene-divinylbenzene polymer, phenol-formalin resin, acrylate resin, and methacrylate resin. Yes, for example, commercial names manufactured by Rohm & Haas, Amberlite XAD-1, Amberlite XAD-2, Amberlite XAD-4, Amberlite XAD-7, Amberlite XAD-8, Amberlite XAD- 11, Amberlite XAD-12, Mitsubishi Kasei Kogyo Co., Ltd. product name Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, manufactured by IMACTI Product name of Immacy Syn-42, Immacy Syn-4 4 and Immacy Syn-46.

  When the reaction liquid in which the 4-alkylresorcinol glycoside of the present invention is produced is passed through, for example, a column filled with the porous synthetic adsorbent as described above, the 4-alkylresorcinol glycoside and a relatively small amount of the glycoside are synthesized. While unreacted 4-alkylresorcinols are adsorbed to the porous synthetic adsorbent, most of the glycosyl compounds that coexist in large quantities flow out without being adsorbed.

  If necessary, after completion of the reaction of glycosyltransferase and before contacting with the porous synthetic adsorbent, for example, the reaction solution is heated to remove insoluble matter, or magnesium aluminate silicate, Treat with magnesium aluminate to adsorb and remove proteinaceous substances in the reaction solution, or treat with strongly acidic ion exchange resin (H type), medium basic or weakly basic ion exchange resin (OH type), etc. It is also optional to use a combination of purification methods such as desalting.

  As described above, the 4-alkylresorcinol glycoside selectively adsorbed on the porous synthetic adsorbent column and the relatively small amount of unreacted 4-alkylresorcinols are washed with a dilute alkali, water, etc. If a relatively small amount of an organic solvent or a mixed solution of an organic solvent and water, for example, methanol water, ethanol water, or the like is passed, 4-alkylresorcinol glycosides are eluted first, increasing the liquid flow rate or increasing the organic volume. If the solvent concentration is increased, unreacted 4-alkylresorcinols are eluted.

  The elution operation of 4-alkylresorcinol glycosides and unreacted 4-alkylresorcinols with this organic solvent is also a regeneration operation of the porous synthetic adsorbent, so that this porous synthetic adsorbent can be used repeatedly. To.

  The eluate with a high content of 4-alkylresorcinol glycoside is subjected to distillation treatment, and after first distilling off the organic solvent, if concentrated to an appropriate concentration, a syrup-like product containing 4-alkylresorcinol glycoside as a main component is obtained. can get. Furthermore, the powdery product which has 4-alkylresorcinol glycoside as a main component is obtained by drying and pulverizing this by methods, such as freeze-drying and spray-drying.

  In addition, the purification using the porous synthetic adsorbent of the present invention has the advantage that not only the glycosyl compound but also impurities such as water-soluble salts contained in the enzyme reaction solution can be removed at the same time.

  As described above, all of the 4-alkylresorcinol glycosides obtained by a synthesis method or an enzymatic method have higher water solubility, stability, and their own properties than 4-alkylresorcinols themselves. Because it maintains its physiological activity, it has excellent antibacterial and whitening effects. This physiological activity is expressed by the fact that the glycoside of 4-alkylresorcinol is decomposed into 4-alkylresorcinols and sugars on the skin surface or in the skin, and the function of 4-alkylresorcinol is reduced. It is thought that it is demonstrated. A mixture of three types of 4-alkylresorcinol glycosides differing in the mode of attachment of the glycosyl group to the hydroxyl group of 4-alkylresorcinol can be isolated as they are, or individually or in combination. The composition of the present invention can be produced. Preference is given to those in which only the hydroxyl group at the 1-position is glycosylated. In addition, these 4-alkylresorcinol glycosides can be treated with an alkali, converted into a salt, and contained as a salt in the composition. Preferred salts include, for example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium and magnesium, organic amine salts such as ammonium salt, triethanolamine salt and triethylamine salt, lysine salt and arginine salt. Preferred examples include basic amino acid salts such as

  In the present specification, the term “whitening” is used to include not only the positive effect of whitening the skin but also the preventive effect of suppressing skin blackening. For example, it includes not only the effect of improving pigmentation such as spots and freckles but also the effect of suppressing pigmentation.

  The blending amount of 4-alkylresorcinol glycosides or salts thereof in the composition of the present invention is limited as long as the physiological functions such as antibacterial, whitening and antioxidant properties of 4-alkylresorcinol glycosides are exhibited. Usually, 0.001% to 10% is used, 0.05% to 5% is preferable, 0.1% to 2% is more preferable, and 0.05 to 1% is particularly desirable. If it is this range, 4-alkyl resorcinol glycoside can be mix | blended easily and there exists an outstanding effect, especially whitening and an antioxidant effect. If it is less than 0.001%, the physiological function of the 4-alkylresorcinol glycoside is not exerted, and if it exceeds 10%, physical properties such as occurrence of starch or coloring due to the compounding prescription or decrease in feeling of use due to the formulation Problems may occur.

  The composition containing the 4-alkylresorcinol glycoside of the present invention refers to pharmaceuticals, quasi drugs, cosmetics, miscellaneous goods and the like. Among these, 4-alkylresorcinol glycosides have an excellent antibacterial action, whitening action, antioxidant action, etc., and therefore can be advantageously blended in a composition for external use for the purpose of whitening. Among these, it is preferable to apply to cosmetics, especially, quasi drugs. For quasi-drugs, it is safer and more effective to display the effect, precautions for use, and the fact that it is a quasi-drug, and clarify the usage. It is preferable for the performance. Such a medicinal effect may be determined according to the characteristics of the active ingredient blended in the external preparation for skin. For example, when ascorbic acids, arbutin and tranexamic acids are included, whitening action based on melanin production inhibitory effect When glycyrrhetinic acids such as dipotassium glycyrrhizinate and stearyl ester of glycyrrhetinic acid are contained, it is preferable to display an anti-inflammatory action. In addition, 4-alkylresorcinol glycoside, which is an essential component of this external preparation for skin, has a whitening action based on a melanin production inhibitory action and an anti-inflammatory action, and therefore suppresses melanin production using such a compound as an active ingredient. It is also possible to display a whitening action based on the action and an anti-inflammatory action.

  In these compositions, as long as the effects of the present invention are not impaired as needed, the components usually used in preparations such as cosmetics, quasi drugs, and pharmaceuticals, that is, water (purified water, hot spring water) are used. ), Alcohols, oils, surfactants, metal soaps, gelling agents, powders, water-soluble polymers, carbohydrates, film-forming agents, resins, UV protection agents, inclusion compounds, antibacterial agents , Preservatives, fragrances, pigments, deodorants, salts, pH adjusters, fresheners, animal and microbial extracts, plant extracts, blood circulation promoters, anti-inflammatory agents, astringents, antiseborrheic agents, whitening Agents, active oxygen scavengers, cell activators, humectants, chelating agents, keratolytic agents, enzymes, hormones, vitamins, minerals, and the like can be added. Specific examples thereof include the following components.

  Alcohol is used for the purpose of dissolution, refreshing feeling, antiseptic, moisturizing and the like. Examples of alcohols include lower alcohols such as ethanol and isopropanol, and glycerin, diglycerin, polyglycerin, ethylene glycol, diethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, pentylene glycol, polyethylene glycol, and isopentyldiol. And the like, and sugar alcohols such as sorbitol, maltitol, maltotriitol, maltotetriitol, and maltopentitol.

  The oil agent is used as a constituent component of the base material for the purpose of improving usability or feeling of use. Any of those used in normal cosmetics can be used, such as whether it is a natural oil or a synthetic oil, or whether it is a solid, semi-solid, liquid, etc. It doesn't matter about the properties. As the oil agent, hydrocarbons, waxes, fatty acids, higher alcohols, ester oils, silicone oils, fluorine oils and the like can be used. More specifically, synthetic ester oil such as cetyl tri-2-ethylhexanoate, pentaerythritol tetra-2-ethylhexanoate, corn oil, olive oil, rapeseed oil, cottonseed oil, coconut oil, palm oil, liquid lanolin, Hardened palm oil, hardened oil, mole, hardened castor oil, olive oil, castor oil, jojoba oil, mink oil, macadamia nut oil, apricot oil, persic oil, safflower oil, sunflower oil, avocado oil, medhome oil, camellia Oils derived from plants and animals such as oil, almond oil, sesame oil, sesame oil, borage oil, and shea fat; waxes such as beeswax, carnauba wax, candelilla wax, ibotarou, lanolin, reduced lanolin, hard lanolin, jojoba wax, and gayrow; Pristane, ozokerite, liquid paraffin, squalane, petrolatum, ceresin Hydrocarbons such as microcrystalline wax; higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid; cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, Higher alcohols such as octyldodecanol, myristyl alcohol, cetostearyl alcohol; cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate, Di-2-ethylhexanoic acid ethylene glycol, dicapric acid neopentyl glycol, di-2-heptyl undecanoic acid glycerin, tri-2-ethylhexyl Synthetic ester oils such as glyceryl acid, tri-2-ethylhexanoic acid trimethylolpropane, triisostearic acid trimethylolpropane, tetra-2-ethylhexanoic acid pentane erythritol; dimethylpolysiloxane, methylphenylpolysiloxane, diphenylpoly Chain polysiloxane such as siloxane; Cyclic polysiloxane such as octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, dodecamethylcyclohexanesiloxane; amino-modified polysiloxane, polyether-modified polysiloxane, alkyl-modified polysiloxane, fluorine-modified polysiloxane Oils such as silicone oil such as modified polysiloxane such as fatty acid soap (sodium laurate, sodium palmitate, etc.), potassium lauryl sulfate, alkyl Anionic surfactants such as trisulfamine trisulfur ether; cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride and laurylamine oxide; imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazoli) Amphoteric surfactants such as nium hydroxide-1-carboxyethyloxy disodium salt), betaine surfactants (alkyl betaine, amide betaine, sulfobetaine, etc.), acylmethyl taurine; sorbitan fatty acid esters (sorbitan mono) Stearates, sorbitan sesquioleate, etc.), glycerin fatty acids (glyceryl monostearate, etc.), propylene glycol fatty acid esters (propylene glycol monostearate, etc.), hardened castor oil derivatives, glycerin al Ether, POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbite fatty acid esters (POE-sorbitol monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoiso) Stearates, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE2-octyldodecyl ether, etc.), POE alkyl phenyl ethers (POE nonylphenyl ether, etc.), pluronic types, POE / POP alkyl ethers (POE / POP2-decyltetradecyl ether, etc.), Tetronics, POE castor oil / hardened castor oil derivative (POE castor oil) Nonionic surfactants such as POE hydrogenated castor oil, sucrose fatty acid ester, alkyl glucoside; polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol, sorbitol, xylitol, maltitol, propylene glycol, dipropylene glycol , Diglycerin, isoprene glycol, 1,2-pentanediol, 2,4-hexanediol, 1,2-hexanediol, 1,2-octanediol and other polyhydric alcohols; sodium pyrrolidonecarboxylate, lactic acid, sodium lactate Moisturizing components such as mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous silicic acid (silica), aluminum oxide, barium sulfate, etc .; surface Processed Bengara, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, bitumen, titanium oxide, zinc oxide inorganic pigments; surface treated, mica titanium, fish phosphorus foil, bismuth oxychloride Pearl agents such as Red 202, Red 228, Red 226, Yellow 4, Blue 404, Yellow 5, Red 505, Red 230, Red 223, Orange Organic dyes such as 201, red 213, yellow 204, yellow 203, blue 1, green 201, purple 201, red 204; polyethylene powder, polymethyl methacrylate, nylon powder, organopolysiloxane Organic powders such as elastomers; vitamin A or its derivatives, vitamin B6 hydrochloride, vitamin B6 tripalmitate, vitamin B6 dioctanoate, vitamin B2 Is a derivative thereof, vitamin B12 such as vitamin B12, vitamin B15 or a derivative thereof; vitamin E such as α-tocopherol, β-tocopherol, γ-tocopherol, vitamin E acetate, vitamin D, vitamin H, pantothenic acid, pantethine Preferred examples include vitamins such as pyrroloquinoline quinone and vitamin P, and derivatives thereof.

  Usable UV protection agents include paraaminobenzoic acid UV absorbers, anthranilic acid UV absorbers, salicylic acid UV absorbers, cinnamic acid UV absorbers, benzophenone UV absorbers, and sugar UV absorbers. be able to. More specifically, paraaminobenzoic acid and its ester, octyl salicylate, homomenthyl salicylate, methyl 2,5-diisopropylcinnamate, cinoxalate, diparamethoxycinnamate mono-2-ethylhexanoate glyceryl, dihydroxydimethoxybenzophenone, Sodium dihydroxydimethoxybenzophenone disulfonate, dihydroxybenzophenone, 1- (3,4-dimethoxyphenyl) -4,4-dimethyl-1,3-pentanedione, dimethicodiethylbenzomalonic acid and its esters, dimethoxybenzylidene dioxoimidazolidine 2-ethylhexyl propionate, tetrahydroxybenzophenone, 2,4,6-tris “-(2-ethylhexyloxycarbonyl) anilino” -1,3,5-triazine, Methyl bis (trimethylsiloxy) silylisopentyl methoxycinnamate, amyl paradimethylaminobenzoate, 2-ethylhexyl paradimethylaminobenzoate, diethylaminohydroxybenzohexyl benzoate, paramethoxycinnamate mixture, paramethylcinnamate 2- Ethylhexyl, 2-hydroxy-4-methoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and its trihydrate, hydroxymethoxybenzophenone sodium sulfonate, 2- (2'-hydroxy-5'-t-octylphenyl) benzotriazole, 4- Methoxy-4′-t-butyldibenzoylmethane, phenylbenzimidazolesulfonic acid, 4-tert-butyl-4-methoxydibenzoylmethane, 2-cyano-3,3- Phenylprop-2-enoic acid 2-ethylhexyl ester (octocreline), 4- (2-β-glucopyranosyloxy) propoxy-2-hydroxybenzophenone, terephthalylidene dicamphulsulfonic acid, ferulic acid, 2,2'- Examples thereof include methylene bis (6- (2Hbenzotriazol-2-yl) -4- (1,1,3,3-tetramethylbutyl) phenol, metrizol trisiloxane, titanium oxide, and zinc oxide.

  Water-soluble polymers and carbohydrates are used for stabilizing the system, improving usability or feeling of use, and also for obtaining a moisturizing effect. Water-soluble polymers that can be used include plant polymers such as pullulan, carrageenan, pectin, agar, locust bean gum, cellulose polymers such as methylcellulose, carboxymethylcellulose, and hydroxypropylcellulose, and alginic acid polymers such as sodium alginate. Examples include molecules and vinyl polymers such as carboxyvinyl polymer. Examples of carbohydrates include maltose, maltotriose, maltotetraose, trehalose, carbohydrate derivatives of trehalose, and cyclic carbohydrates such as cyclic tetrasaccharides.

  Examples of antiseptics and antibacterial agents include benzoic acid, sodium benzoate, paraoxybenzoic acid ester, benzalkonium chloride, phenoxyethanol, isopropylmethylphenol and the like.

  The 4-alkylresorcinol glycoside of the present invention has an excellent whitening effect alone, but in order to further improve the effect, a conventionally known whitening agent may be used in combination with the composition of the present invention. . The whitening agent is used for the purpose of preventing skin blackening caused by sunburn or the like, spots such as spots or freckles caused by pigmentation. Other whitening agents that can be used include arbutin, linoleic acid, 5,5'-dipropylbisphenyl-2,2'-diol, vitamin C and derivatives thereof, vitamin E and derivatives thereof, glycyrrhizic acid and derivatives thereof , Tranexamic acid and its derivatives, tetrahydrocurcuminoid, placenta extract, indigo extract, chamomile extract, licorice extract, ages extract, oxon extract, seaweed extract, cucumber extract, quette extract, gokahi extract, Rice bran extract, wheat germ extract, saicin extract, hawthorn extract, sun pens extract, white lily extract, peonies extract, sempukuka extract, soybean extract, tea extract, molasses extract, juniper extract, grape Extract, Hop extract, Mikaika extract, Mokka extract, Yukinoshita extract, Yoku Examples include inine extract and royal jelly extract.

  Anti-inflammatory agents are used for the purpose of suppressing inflammation such as hot flashes and erythema after sunburn. Anti-inflammatory agents include sulfur and derivatives thereof, glycyrrhizic acid and derivatives thereof, glycyrrhetinic acid and derivatives thereof, lepocarcinin chloride, aloe extract, artea extract, ashitaba extract, arnica extract, indigo extract, inchinkou extract, Nettle extract, buckwheat extract, hypericum extract, chamomile extract, licorice extract, goldfish extract, watercress extract, comfrey extract, salvia extract, lion extract, perilla extract, birch extract, gentian An extract etc. can be mentioned.

  The cell activator is used for the purpose of improving rough skin. Examples of the cell activator that can be used in the composition of the present invention include caffeine, chicken crown extract, shell extract, shell extract, royal jelly, silk protein and its degradation product or derivatives thereof, lactoferrin or its degradation product, Mucopolysaccharides such as chondroitin sulfate and hyaluronic acid or salts thereof, collagen, yeast extract, lactic acid bacteria extract, bifidobacteria extract, fermentation metabolic extract, ginkgo biloba extract, barley extract, assembly extract, peanut extract, Examples thereof include carrot extract, rosemary extract, glycolic acid, citric acid, lactic acid, malic acid, tartaric acid, organic acids such as succinic acid, and derivatives thereof.

  The active oxygen scavenger is used for the purpose of suppressing oxidative damage such as lipid peroxide generation. Examples of active oxygen scavengers include superoxide dismutase, mannitol, quercetin, catechin and derivatives thereof, rutin and derivatives thereof, bopi extract, yashajitsu extract, melissa extract, rahan extract, vitamins such as retinol and carotenoid, and the like Derivatives thereof, vitamin Bs and derivatives thereof such as thiamine, riboflavin, pyridoxine, nicotinic acid, vitamin C and derivatives thereof, vitamin Ps and derivatives thereof such as rutin, hesperidin, naringin, vitamin E compounds such as tocopherol and derivatives thereof, Examples include dibutylhydroxytoluene and butylhydroxyanisole.

  Examples of humectants include proteins such as elastin and keratin or derivatives thereof, hydrolysates and salts thereof, amino acids such as glycine, serine, aspartic acid, glutamic acid, arginine and theanine and derivatives thereof, sorbitol, erythritol, trehalose and Derivatives thereof, inositol, glucose, maltose, sucrose and derivatives thereof, dextrin, cyclodextrin, cyclic tetrasaccharides and derivatives thereof, saccharides such as honey, D-pantenol and derivatives thereof, urea, phospholipid, ceramide, urene extract , Ginseng extract, gypsophila extract, gypsophila extract, gypsophila extract, dokudami extract, hamamelis extract, bodaige extract, maronier extract, quince extract and the like.

  The form of the external composition for skin of the present invention is not particularly limited. For example, lotion, essence, milky lotion, cream, lotion, pack, cosmetic liquid, cleaning agent, makeup cosmetics, hair care cosmetics, bath preparation, dispersion liquid, An ointment, a liquid agent, an aerosol agent, a patch, a poultice, a liniment, etc. can be mentioned. In addition to the treatment and prevention of diseases, the purpose of use can also be applied to applications that are generally known for external preparations for skin, such as basic skin care and makeup. Particularly preferred are emulsified essence preparations or cream preparations that can fully utilize the effects of the present invention. Moreover, there is no restriction | limiting in particular in the manufacturing method, It can manufacture by a conventional method.

  EXAMPLES Next, although an Example is given and this invention is demonstrated further in detail, this invention is not limited to the following Example at all.

  1.9 g of 4-cyclohexylresorcinol was weighed, and 2 ml of bistrimethylsilylacetamide was dissolved in 10 ml of acetonitrile and added thereto, stirred for 30 minutes at room temperature, reacted, distilled off, and 100 ml of acetonitrile was added again. In addition, 200 mg of silver peroxide and 5 g of pentaacetylglucose were dissolved and dispersed in 20 ml of acetonitrile and added dropwise under ice cooling. After the dropwise addition, the ice bath was removed, the temperature was gradually returned to room temperature, then heated and refluxed for 2 hours. After cooling to room temperature, it was poured onto ice with sodium bicarbonate, the reaction was stopped, extracted with 500 ml of ethyl acetate, washed twice with 100 ml of water, the ethyl acetate phase was dried over anhydrous sodium sulfate, concentrated and then concentrated on a silica gel column. Purified by chromatography (elution solvent; chloroform: methanol = 100: 0 → 90: 10 → 70: 30), 4-cyclohexyl-1- (tetraacetylglucosyl) resorcinol and 4-cyclohexyl-3- (tetraacetylglucosyl) Resorcinol and 1,3-bis (tetraacetylglucosyl) -4-cyclohexylresorcinol were obtained. These were each treated with an ammonia saturated methanol solution for 5 hours to give 421 mg of 4-cyclohexyl-1-glucosylresorcinol (Compound 1), 345 mg of 4-cyclohexyl-3-glucosylresorcinol (Compound 2) and 1,3-bisglucosyl-4- 95 mg of cyclohexylresorcinol (compound 3) was obtained.

  As in Example 1, 4-cyclohexylresorcinol was replaced with 4-cyclopentylresorcinol and treated to give 396 mg of 4-cyclopentyl-1-glucosylresorcinol (compound 4), 223 mg of 4-cyclopentyl-3-glucosylresorcinol (compound 5) and 119 mg of 1,3-bisglucosyl-4-cyclopentylresorcinol (Compound 6) was obtained.

  In the same manner as in Example 1, 4-cyclohexylresorcinol was changed to 4- (2-phenylethyl) resorcinol and treated to give 287 mg of 1-glucosyl-4- (2-phenylethyl) resorcinol (Compound 7), 3-glucosyl- 181 mg of 40 (2-phenylethyl) resorcinol (Compound 8) and 76 mg of 1,3-bisglucosyl-4- (2-phenylethyl) resorcinol (Compound 9) were obtained.

  In the same manner as in Example 1, 4-cyclohexylresorcinol was changed to 4- (1-methylpropyl) resorcinol and treated to give 321 mg of 1-glucosyl-4- (1-methylpropyl) resorcinol (Compound 10), 3-glucosyl- There were obtained 239 mg of 40 (1-methylpropyl) resorcinol (Compound 11) and 105 mg of 1,3-bisglucosyl-4- (1-methylpropylresorcinol (Compound 12).

  In the same manner as in Example 1, 4-cyclohexylresorcinol was replaced with 4- (1-ethylpropyl) resorcinol and treated to give 291 mg of 4- (1-ethylpropyl) -1-glucosylresorcinol (Compound 13), 4- (1 -Ethylpropyl) -3-glucosylresorcinol (Compound 14) 204 mg and 1,3-bisglucosyl-4- (1-ethylpropylresorcinol (Compound 15) 83 mg were obtained.

  In the same manner as in Example 1, 4-cyclohexylresorcinol was changed to 4- (1-isopropyl-2-methylpropyl) resorcinol and treated to give 1-glucosyl-4- (1-isopropyl-2-methylpropyl) resorcinol (compound 16) 168 mg, 3-glucosyl-40 (1-isopropyl-2-methylpropyl) resorcinol (compound 17) 118 mg and 1,3-bisglucosyl-4- (1-isopropyl-2-methylpropyl) resorcinol (compound 18) 64 mg was obtained.

<Test Example 1: Stability of 4-alkylresorcinol glycoside>
Similar to the reference example, the stability of the compounds 1 to 18 prepared in Examples 1 to 6 was examined. The results are shown in Table 2.

  As is clear from Table 2, it can be seen that the stability was improved by glycosylation.

<Examination of pharmacological activity of resorcinol glycosides>
The melanin production inhibitory effect was investigated by melanoma B-16. That is, melanoma B-16 cells in the logarithmic growth phase were trypsinized and then added to an FBS (fetal calf serum) -containing MEM medium to a concentration of 1.5 × 10 ³ cells / ml, and melanoma B-16 An FBS-containing MEM suspension was made. 10 ml of this suspension was dispensed into five culture bottles, placed in a carbon dioxide incubator at 37 ° C. and 5% by volume CO ₂ concentration, and cultured for 2 days. Thereafter, the 4-culture resorcinol (control), Compound 1, Compound 2 and Compound 3 were each added to 0.001% (final concentration) of 4 culture bottles in MEM containing 10% by volume DMSO / FBS. 5 ml each was added together with the medium solution. To the remaining one, 5 ml of 10 volume% DMSO / FBS-containing MEM medium solution was added as a control. These bottles were cultured under conditions of 37 ° C. and 5% by volume carbon dioxide for 2 days. Thereafter, 15 ml of FBS-containing MEM medium was added to each culture bottle, and 4-cyclohexylresorcinol, Compound 1, Compound 2, and Compound 3 were added in the same manner as above, and vehicle was added to the remaining one. This was further cultured for 2 days. After completion of the culture, the medium was removed from each bottle, washed with PBS (phosphate buffered saline), and then treated with trypsin to detach the cells from the culture bottle to obtain a cell suspension. The cell suspension was centrifuged to collect cells. The number of the obtained cells and the degree of melanin pigmentation were observed and visually observed according to the following criteria to evaluate cytotoxicity and melanin production inhibitory action. The results are shown in Table 3.
[Evaluation criteria for cytotoxicity]
+: The number of cells is considerably smaller than that of the control and cytotoxic.
±: The number of cells is slightly smaller than that of the control, but it is difficult to determine cytotoxicity.
-: There is almost no difference in the number of cells compared to the control, and no cytotoxicity is observed.
[Inhibition of melanin production]
+: Clearly whitened as compared with the control, and has an inhibitory effect on melanin production.
±: Slightly whitened from the control, and a slight melanin production inhibitory effect is observed.
−: Almost the same as control, and no melanin production inhibitory effect is observed.

  As is apparent from Table 3, the glycoside which is the compound of the present invention has the same melanin production inhibitory action as aglycone. Moreover, it turns out that cytotoxicity falls by glycosylation.

<Preparation of 4-n-butylresorcinol glycoside>
4-n-Butyl resorcinol was prepared according to the method described in Example 1 of British Patent 1,581,428. 4 g of this 4-n-butylresorcinol and 80 g of α-cyclodextrin (hereinafter sometimes abbreviated as “α-CD”) were dissolved in 800 mL of 50 mM acetate buffer (containing pH 5.5, ₂ mM CaCl ₂ ). Thereafter, 500 units / g-α-CD of Bacillus stearothermophilus-derived cyclomaltodextrin glucanotransferase (available from Hayashibara Biochemical Laboratories Co., Ltd.) was added and reacted at 40 ° C. for 24 hours. After stopping the reaction by heating at 100 ° C. for 10 minutes, 80 mL of 1M acetate buffer (pH 4.5) was added, and 10 units / g-α-CD of glucoamylase (Seikagaku Corporation) was added, followed by 50 ° C. For 20 hours. After stopping the reaction by heating at 100 ° C. for 10 minutes, a column (7 cmφ × 39 cm, 1.5 L) packed with a porous synthetic adsorbent (Mitsubishi Kasei Co., Ltd., trade name “Diaion HP-20”) was used. Provided. The column was washed with water, and 35% ethanol was passed through to elute the fraction adsorbed on the column. The obtained fraction was concentrated to dryness, suspended and dissolved in 96% ethanol, and then insoluble carbohydrates were removed. 2. After distilling off the solvent of the supernatant, it is again dissolved in water, freeze-dried, and a powder preparation containing a reaction product by glycosyltransferase (hereinafter sometimes abbreviated as “reaction product”). 21 g was obtained. Using a portion of the obtained powder sample, it was subjected to high performance liquid chromatography (hereinafter sometimes abbreviated as “HPLC”), and liquid chromatogram mass spectrometry (hereinafter referred to as “LC / MS”) of the eluted fraction. ”Analysis and ultraviolet (hereinafter sometimes abbreviated as“ UV ”) absorption spectrum analysis. These analyzes were performed under the following conditions.
<HPLC>
Equipment: CR-4A and LCD-10AD (sold by Shimadzu Corporation)
Column: CAPCELLPAK C18 AG120 4.6 mmφ × 2
50mm (manufactured by Shinwa Kako Co., Ltd.)
Mobile phase: acetonitrile / water / acetic acid (35: 65: 0.2)
Flow rate: 0.5mL / min
Temperature: 40 ° C
<UV>
Equipment: SPD-10AVP (Sold by Shimadzu Corporation)
Wavelength: UV280nm
<LC / MS>
Equipment: LCQ Advantage (sales by Thermo Electron Co., Ltd.)
Detection condition:
ESI, negative mode, m / z 50-2000
Sheath Gas Flow: 40
Ion Spray Voltage: 5kV
Capillary Temp. : 275 ° C
Capillary Voltage: -7.0V
Tube Lens Offset: -10.0V

  The aqueous solution of the lyophilized preparation containing the reaction product shows almost the same absorption spectrum as the raw material 4-n-butylresorcinol (FIG. 1), and the resorcinol skeleton part of the reaction product by glycosyltransferase has There were no major changes. Further, when this sample was subjected to HPLC, as shown in FIG. 2, the retention times were two kinds having absorption at UV 280 nm (having a resorcinol skeleton) at positions of about 7.34 minutes and about 9.42 minutes. The peak of the reaction product was observed (hereinafter referred to as “reaction product a” and “reaction product b”). In the MS analysis of these two peak portions, as is apparent from FIG. 3 showing the relationship between the value obtained by dividing the mass of the ion by the charge (M / Z) and the ionic strength, both are 387 acetate ion-added molecules. Since ions were detected and the molecular weight of acetate ions was 59, it was found that the molecular weights of the two kinds of reaction products were both 328. The content of these reaction products in 3.21 g of the powder sample is determined by measuring the elution peak area of each reaction product shown in the chart showing the HPLC elution pattern based on the absorbance at UV 280 nm, and the UV 280 nm mole of 4-n-butylresorcinol. From the extinction coefficient (calculated on the assumption that 4-n-butylresorcinol and its glycoside are the same as the molar extinction coefficient of UV280 nm) since the carbohydrate part of the glycoside has no ultraviolet absorption. And 1.87 g (Table 4).

<Test Example 2: Determination of structure of reaction product>
1 g of the dry powder preparation containing the reaction product prepared in Example 7 was dissolved in a solution of acetonitrile / water / acetic acid (= 30/70 / 0.2), and the solution used for this dissolution was used as the mobile phase. The D-ODS-5 S5 (available from YMC) column chromatography used was used for fractionation, and two types of reaction product a (purity 99% or more) and reaction product b (purity 96.8%) were obtained. 0.25 g and 0.56 g were obtained. Using some of these reaction products, NMR analysis was performed under the following conditions to determine the respective structures.
<NMR apparatus>
Equipment: JNM-AL300 (JEOL stock sales)
Solvent: D ₂ O
Reference substance: sodium trimethylsilylpropionate-2,2,3,3-d ₄
Magnetic field strength: ¹ H-NMR 300.4 MHz
¹³ C-NMR 75.45 MHz

<NMR analysis>
Chemical shifts (FIGS. 4 to 13) by NMR analysis of reaction products a and reaction products b shown below, and hydrogen of the benzene ring are hydroxyl groups (1st and 3rd positions), O-glucosyl groups (3rd position, With reference to the calculated chemical shift value when substituted with 1-position) and n-butyl group (4-position), all signals of NMR analysis of reaction product a and reaction product b were assigned. The results are shown in Table 5.

<Analysis of reaction product a by NMR>
¹ H-NMR and ¹³ C-NMR spectra are shown in FIGS. As is clear from FIG. 4, in ¹ H-NMR, three benzene protons are located at a position where the chemical shift is 6 ppm to 7 ppm, and six protons other than the sugar hydroxyl group are located at a position where the chemical shift is 3 ppm to 6 ppm. Nine butyl group protons were observed at a chemical shift of 0.5 ppm to 3 ppm. As is clear from FIG. 5, in the ¹³ C-NMR spectrum, 6 signals of benzene carbon (positions 1 and 3 are almost the same position) at a position where the chemical shift is 100 ppm to 160 ppm, and the chemical shift is 60 ppm to 100 ppm. Six sugar carbon signals were observed at the position, and four butyl group carbon signals were observed at the chemical shift positions of 10 ppm to 35 ppm. In addition, from the chemical shift δ (δ _C ) value of each carbon atom shown in Table 2, the glucose residue is of a pyranose type, and the chemical shift δ _C of the anomeric carbon to the chemical shift δ _C 97.1 ppm is δ (δ _H ) The signal of the anomeric proton was observed at 5.52 ppm (FIGS. 4 and 6), and the chemical shift at the 1 ″ position of the glucosyl group (δ _C 97.1 ppm, δ _H 5.52 ppm (1H, d, J = From 3.3 Hz)) (Table 5), it was considered that the glucose residue was α-bonded. Furthermore, geminal non-equivalence was observed for the 1'-position proton of the butyl group (FIGS. 5 and 7).

<Analysis of reaction product b by NMR>
¹ H-NMR and ¹³ C-NMR spectra are shown in FIGS. The NMR spectrum of the reaction product b is the same as that of the reaction product a. In ¹ H-NMR, three benzene protons are located at a position where the chemical shift is 6 ppm to 7 ppm, and protons other than the sugar hydroxyl group are located at the positions 3 ppm to 6 ppm. 9 and 9 butyl group protons were observed at a chemical shift of 0.5 ppm to 3 ppm (FIG. 8), but unlike the reaction product a, geminal non-equivalent of the 1'-position proton of the butyl group Sex was hardly recognized (FIGS. 8 and 10). ^{The 13} C-NMR spectrum was very similar to the reaction product a. Like the reaction product a, glucose is α-bonded from the chemical shift at the 1 ″ position of the glucosyl group (δ _C 98.0 ppm, δ _H 5.30 ppm (1H, d, J = 3.7 Hz)). (Fig. 9, Fig. 11 and Table 5). In addition, when benzene carbon (2nd and 6th positions) in which correlation with the glucosyl group 1 ″ proton is observed in the remote C—H COZY, the butyl group 1 ′ position proton and the remote coupling are found. It was different from the recognized benzene carbon (3rd to 5th positions) (FIGS. 12 and 13).

  From these analysis results, it was considered that in the reaction product b, glucose was transferred to the 1st position instead of the 3rd position. Therefore, it was considered that glucose was transferred to the 3-position in reaction product b and reaction product a having a different NMR spectrum. In addition, the non-equivalence of the butyl group geminal proton occurs only in the reaction product a. In the reaction product a, a glucose residue is present adjacent to the butyl group. It is considered that the binding site of the glucose residue of the reaction product a coincides with the 1st position. Therefore, these test results and the analysis results are as follows: the reaction product a is 3-O-α-D-glucopyranosyl-4-n-butylresorcinol represented by Chemical Formula 1, and the reaction product b is represented by Chemical Formula 2. It has a structure of -O-α-D-glucopyranosyl-4-n-butylresorcinol.

  Although specific MS analysis data is not shown, in the LC / MS analysis results of the HPLC elution fraction using the CAPCELLPAK C18 AG120 column shown in FIG. 2, the retention time is 5.08 min to 5.33 3 fractions eluted around 5 minutes, 5.95 minutes to 6.20 minutes, and 8.29 minutes have absorption at UV 280 nm (having resorcinol skeleton) and mass addition of 549 acetate ions The detection of molecular ions revealed that these fractions contained three types of reaction products having a mass of 490 and different hydrophobic strengths. The mass of these reaction products corresponds to the molecular weight of butylresorcinol in which two molecules of glucose are bound to butylresorcinol, and, as shown in Chemical Formula 1, 4-n-butylresorcinol by glycosyltransferase is used. Since the transfer site of the glucose residue to the OH group is only the 1st and 3rd OH groups, these three kinds of mass 490 reaction products are not completely digested with glucoamylase. 1-O-α-D-maltosyl-4-n-butylresorcinol, in which a maltosyl group is α-bonded to the 1-position OH group of resorcinol, and two molecules of the maltosyl group are α -Bound 3-O- [alpha] -D-maltosyl-4-n-butylresorcinol and one OH group each α-bonded to both OH groups It was judged to be any of 1,3-O-α-D-glucopyranosyl-4-n-butylresorcinol. Therefore, this result is less than the amount of glycosides in which glucose is bound to any one of the OH groups in the transglycosylation reaction to 4-n-butylresorcinol using cyclomaltodextrin glucanotransferase. However, it shows that glycosides with glucose residues bound to both OH groups are also produced.

<Test Example 3: Solubility test>
10 mg of 4-n-butylresorcinol (hereinafter referred to as “4-n-butylresorcinol used in Example 7”) used as a substrate for preparing glycosides in Example 7, and D- in Experiment 1 10 mg of 1-O-α-D-glucopyranosyl-4-n-butylresorcinol (reaction product b) prepared by column chromatography using ODS-5 S5 and 3-O-α-D-glucopyranosyl-4 Each of 10 mg of n-butylresorcinol (reaction product a) was added with deionized water, stirred with a vortex mixer for 2 to 3 minutes, and allowed to stand for insoluble matter when left standing. The maximum concentration was determined and taken as the solubility of each compound. The results are shown in Table 6.

  As apparent from Table 6, the solubility of the two types of 4-n-butylresorcinol glycosides was twice that of 4-n-butylresorcinol. Thereby, it was confirmed that the solubility with respect to water improves by carrying out glycosylation of 4-n-butyl resorcinol. Further, no difference in solubility depending on the binding position of glucose was observed.

<Test Example 4: Examination of pharmacological activity>
The reaction product-containing lyophilized powder prepared in Example 7 (in addition to 1-O-α-D-glucopyranosyl-4-n-butylresorcinol and 3-O-α-D-glucopyranosyl-4-n-butylresorcinol , Containing a small amount of 4-n-butylresorcinol glycoside in which two or more glucosyl residues are bonded to 4-n-butylresorcinol: hereinafter referred to as “4-n-butylresorcinol glycoside-containing powder”). Three kinds of 4-n-butylresorcinol glycoside preparations of 1-O-α-D-glucopyranosyl-4-n-butylresorcinol and 3-O-α-D-glucopyranosyl-4-n-butylresorcinol; Using 4-n-butylresorcinol used in Example 7, a test for confirming the whitening effect was performed as follows. That is, according to Test Example 1, mouse B16 melanoma cells are cultured, and after removing the culture supernatant, either 4-n-butylresorcinol or three types of 4-n-butylresorcinol glycoside preparations are added. Then, a medium prepared such that 4-n-butylresorcinol or 4-n-butylresorcinol glycoside has a concentration shown in Table 7 was added, followed by further culturing for 72 hours. The cells were collected, the cells were packed into a capillary tube, the color tone was observed, the degree of whitening action based on the melanin production inhibitory action was determined according to the following criteria, and the results are shown in Table 7. As a positive control, the whitening effect was similarly determined using kojic acid (2.5 mM) used as a skin whitening agent in a skin external preparation. In addition, a portion of the collected cells is stained with a dye that stains only living cells (Alama Blue), and the ratio (%) of viable cells stained with the dye that is incorporated into all cells is obtained. ) As shown in Table 7. The concentration of 4-n-butylresorcinol glycoside in the powder containing 4-n-butylresorcinol glycoside used in the following experiment was calculated based on the molar extinction coefficient of resorcinol at UV 280 nm.

[Whitening effect]
Vs. blackness of control without drug-: no change.
±: Slightly whitened.
+: Slightly whitened.
++: A considerable whitening occurred.
+++: Completely whitened.

  As is clear from Table 7, the three types of 4-n-butylresorcinol glycoside preparations are all 0.5 mM or 2.0 mM, equivalent to 2.5 mM kojic acid installed as a positive control substance, or A stronger whitening effect was observed. Further, no difference was observed in the strength of the whitening action among the three types of 4-n-butylresorcinol glycoside preparations used. On the other hand, 4-n-butylresorcinol showed a whitening effect equivalent to three kinds of 4-n-butylresorcinol glycoside preparations at a concentration of 0.5 mM, and a strong whitening effect was observed at a concentration of 1 mM. Since the survival rate of B16 melanoma cells was reduced to 79.7%, this whitening action was considered to be due to the cytotoxicity of 4-n-butylresorcinol. The three types of 4-n-butylresorcinol glycoside preparations have a B16 melanoma cell survival rate of 95.8% to 97.3% even when the concentration is 2.0 mM, and the medium alone (no addition) Since the cell viability was almost the same as that of the cultured cells, and no cytotoxicity was observed, 4-n-butylresorcinol was glycosylated to reduce the whitening effect without reducing the whitening effect. It became clear that it was reduced.

<Test Example 5: Examination of pharmacological activity>
Using the 4-n-butylresorcinol glycoside-containing powder sample prepared in Example 7, a test for confirming the whitening effect of 4-n-butylresorcinol glycoside was performed as follows. That is, by the same method as in Test Example 2, mouse B16 melanoma cells were cultured for 24 hours, the culture supernatant was removed, and a medium containing each test sample to the concentration shown in Table 8 was added. Cultured for 72 hours. The cells were collected, packed in a capillary tube, and the color tone was observed. The degree of whitening effect based on the melanin production inhibitory action was determined according to the following criteria, and the results are shown in Table 8.

[Whitening effect]
Vs. blackness of control without drug-: no change.
±: Slightly whitened.
+: Slightly whitened.
++: A considerable whitening occurred.
+++: Completely whitened.

  As is apparent from Table 8, 4-n-butylresorcinol glycoside-containing powder preparation is composed of L-ascorbic acid sodium salt, potassium 4-methoxysalicylate, kojic acid, arbutin, tranexamic acid, ellagic acid, linoleic acid, When used in combination with chamomile extract and tetrahydrocurcumin, a synergistic whitening effect was observed. This result shows that 4-n-butylresorcinol glycoside is used in combination with L-ascorbic acid sodium salt, potassium 4-methoxysalicylate, kojic acid, arbutin, tranexamic acid, ellagic acid, linoleic acid, chamomile extract, and tetrahydrocurcumin In this case, it is said that it synergistically suppresses the production of melanin and exhibits an excellent whitening effect.

<Test Example 6: Evaluation of antioxidant effect of 4-n-butylresorcinol glycoside: lipid peroxidation inhibitory effect>
To 1.5 mL of a mixed solution of 1% egg yolk lecithin and 0.4% sodium dodecyl sulfate, 0.5 mL of 2.4 mmol / L ferrous sulfate and 0.5 mL of 4.8 mmol / L ascorbic acid solution were added. In addition, a solution containing 4-n-butylresorcinol glycoside having a concentration shown in Table 9 prepared by dissolving the powder containing 4-n-butylresorcinol glycoside prepared in Example 7 in a buffer solution 0.5 mL of any of the above was added and heated in a 37 ° C. water bath for 1 hour. Thereafter, the thiobarbituric acid value (hereinafter referred to as “TBA value”) was measured by a conventional method to determine the degree of peroxidation. Instead of 4-n-butylresorcinol glycoside-containing powder, 0.5 mL of a buffer solution as a blank and 0.10% kojic acid solution as a positive control were added and reacted in the same manner. For the solution preparation, 10 mmol / L Tris-HCl buffer (pH 7.5) was used. Taking the TBA value of the blank as 100, the relative value of the TBA value when 4-n-butylresorcinol glycoside-containing powder or positive control was added was determined, and the inhibition rate of lipid peroxide generation determined by subtracting from 100 ( %) Is shown in Table 9 as the antioxidant effect.

  As is apparent from Table 9, when 4-n-butylresorcinol glycoside-containing powder was added, addition of 0.001% or more suppressed the production of lipid peroxide depending on the amount added, At 0.01%, the inhibition rate was 29.8%, and the same inhibition effect as when 0.1% kojic acid was added (suppression rate 30.0%) was observed. Furthermore, when 0.05% is added, the inhibition rate is increased to 42.5%, and when 0.10% or more is added, a strong inhibitory effect exceeding 50% is observed, and 4-n-butylresorcinol glycoside-containing powder Was confirmed to be superior in antioxidant effect compared to kojic acid. This result shows that by adding 4-n-butylresorcinol glycoside to the skin external preparation, not only a strong antioxidant effect on the skin can be obtained, but also the stability of the skin external preparation can be improved as an antioxidant. It tells you that you can contribute.

  Since these test results show that 4-n-butylresorcinol glycoside has a melanin production inhibitory action and an antioxidant action, the composition for external use with skin is whitening and / or skin beautifying and anti-aging. It is telling that it is effective. Moreover, since 4-n-butylresorcinol glycoside has lower cytotoxicity than 4-n-butylresorcinol, when it is added to a skin external preparation, the irritation to skin is more than 4-n-butylresorcinol. Also tells the low.

<Production of 4-n-butylresorcinol glycoside>
4-n-Butyl resorcinol was prepared according to the method described in Example 1 of British Patent 1,581,428. After dissolving 20 g of 4-n-butylresorcinol in 800 mL of 50 mM acetate buffer (pH 5.5), 400 g of dextrin (Matsuya Chemical Co., Ltd., trade name “Paindex # 1”) was added and stirred. Acetate buffer was added to make a total volume of 4000 mL. Thereto was added 600 units / g-dextrin of cyclomaltodextrin glucanotransferase derived from Bacillus stearothermophilus (sold by Hayashibara Biochemical Laboratories Co., Ltd.) and reacted at 50 ° C. for 24 hours. After the reaction solution was heated at 100 ° C. for 10 minutes to stop the reaction, the reaction solution was filtered, and glucoamylase (EC 3.2.1.3, sold by Seikagaku Corporation) was added to this at 10 units / g-dextrin was added, and the reaction was carried out for 5 hours while maintaining the pH at 4.5 and 55 ° C. The reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was loaded with SV2 onto a column packed with 6 L of a porous synthetic adsorbent, trade name Diaion HP-10 (Mitsubishi Kasei Kogyo Co., Ltd.). After the column was washed with water, 35% ethanol was passed through to elute the glycoside fraction. The obtained fraction was concentrated to dryness, suspended and dissolved in 96% ethanol, and then insoluble matters were removed. The supernatant was concentrated to dryness, dissolved again in water, and then freeze-dried to obtain a powder sample containing 4-n-butylresorcinol glycoside. After dissolving this sample in acetonitrile / water / acetic acid (= 30/70 / 0.2), D-ODS-5 S5 (sold by YMC) column chromatography using the solution used for this dissolution as a mobile phase. By fractionation by chromatography, two kinds of reaction products a (purity 99% or more) and reaction product b (purity 96.8%) were obtained in a yield of 95%.

  This reaction product a and reaction product b were subjected to MS analysis and NMR analysis in the same manner as in Test Example 3. As in Experiment 1, the reaction product a was 3-O-α-D-glucopyranosyl. It was -4-n-butylresorcinol, and it was confirmed that the reaction product b was 1-O-α-D-glucopyranosyl-4-n-butylresorcinol. Further, as in Experiment 1, although the production amount is smaller than that of the reaction product a and the reaction product b, 1,3-O-α-D-glucopyranosyl-4-n is added to the fraction eluted at different positions. -Production of a substance corresponding to butylresorcinol was confirmed.

[Safety test]
A lyophilized powder sample containing 4-n-butylresorcinol glycoside prepared in Example 7, reaction product a and reaction product prepared by column chromatography using D-ODS-5 S5 in Example 7 3 parts by mass of the product b (freeze-dried sample is 3 parts by mass as 4-n-butylresorcinol glycoside), 5 parts by mass of ethanol, 5 parts by mass of glycerol and 87 parts by mass of purified water It melt | dissolved in the mixed solution and prepared the test liquid. Skin condition at the site where any one of these solutions was applied to the inside of the upper arm of 5 volunteers each (15 people in total) in a range of 1 cm × 1 cm 3 times a day for 3 days and then applied for 1 week Was observed. As a control, only the base was applied 3 times a day for 3 days in a 1 cm x 1 cm range adjacent to the area where the test solution was applied, and the skin condition was observed for 1 week from the start date of application. did. In any case where the test solution or the control solution was applied, no changes such as rough skin and redness were observed at the application site, and these three types of 4-n-butylresorcinol glycoside preparations were It was also judged to be a highly safe substance even when blended with topical skin preparations.

<Production of 7 types of alkylresorcinol glycosides with different alkyl groups>
According to the method of Example 7, 4-n-methylresorcinol, 4-n-ethylresorcinol, 4-n-hexylresorcinol, 4-n-heptylresorcinol, 4-n-octylresorcinol, 4-n-decylresorcinol And 5 g of 4-n-eicosylresorcinol and 100 g of dextrin, a transglycosylation reaction was performed, and the reaction solution was treated with a porous synthetic adsorbent (sold by Mitsubishi Kasei Corporation, trade name “ Purified using a column packed with Diaion HP-20 "), saccharides precipitated with 96% ethanol were removed, freeze-dried, and these alkylresorcinol glycoside powder preparations (purity of 85% to 91%) was prepared.

  In addition to the seven alkylresorcinol glycoside powder preparations prepared in Example 9, the same method as in Experiment 3 using the 4-n-butylresorcinol glycoside powder preparation prepared in Example 7 Then, each glycoside was added to mouse B16 melanoma medium in the same manner as in Test Example 4 so as to be 0.5 mM or 2 mM, and the whitening test was carried out. As a result, 4-n− having a concentration of 2 mM was obtained. A strong whitening effect (substantially whitened) was observed with the addition of butylresorcinol glycoside-containing powder. 4-n-methylresorcinol glycoside, 4-n-ethylresorcinol glycoside, 4-n-butylresorcinol glycoside, 4-n-hexylresorcinol glycoside, 4-n-heptylresorcinol glycoside When added to the medium, a fairly strong whitening effect (slightly whitened) was observed at a concentration of 2 mM. In addition, when 4-n-octylresorcinol glycoside, 4-n-decylresorcinol glycoside and 4-n-eicosylresorcinol glycoside were added to the medium, whitening effect was observed. As compared with the case where -n-heptylresorcinol glycoside or an alkylresorcinol glycoside having a lower molecular weight was added, the whitening effect was weak (slightly whitened). At the concentrations used in this experiment, the cell survival rate was 96% to 99% at any alkyl resorcinol glycoside, and no cytotoxicity against B16 melanoma was observed at the concentrations used in the test. It was.

<Lotion>
Using the compounds 1 to 18 prepared in Examples 1 to 6, lotion cosmetics 1 to 18 which are skin external preparations of the present invention were prepared according to the following formulation. That is, the formulation components were solubilized by stirring at 80 ° C. and stirred and cooled to obtain a lotion. Each of these lotions had an excellent whitening effect. In addition, the mixing | blending of each component of the Example which shows the composition which mix | blended the glycoside of this invention shown below is shown by the ratio (%) of each component which occupies for the gross mass of cosmetics.
(Prescription) (%)
(1) 1,3-butanediol 5
(2) 1,2-pentanediol 3
(3) Dipotassium glycyrrhizinate 0.1
(4) L-ascorbic acid-2-glucoside 3
(5) Any one of Compounds 1 to 18 prepared by the methods of Examples 1 to 6
(6) Glycerin 3
(7) Water 84.9

<Lotion>
A lotion was prepared by the conventional method using the following formulation. This product was an excellent cosmetic product that, when applied to the skin, whitens the skin and gives the skin tension and luster. Moreover, since this product contained 4-n-butylresorcinol glycoside, the storage stability was also good.
(Prescription) (%)
(1) Glycerin 5
(2) 1,3-butylene glycol 6.5
(3) Polyoxyethylene sorbitan monolaurate (20E.O.) 1.2
(4) Ethyl alcohol 8
(5) 4-n-butyl resorcinol glycoside-containing powder prepared by the method of Example 7 0.5
(6) Lactic acid 0.05
(7) Sodium lactate 0.1
(8) Paramethoxycinnamic acid-2-ethylhexyl 0.1
(9) Pullulan 0.05
(10) Preservative appropriate amount (11) Fragrance appropriate amount (12) Water appropriate amount

  10 volunteers use this lotion for 3 days a day for 10 days, and replace with 4-n-butylresorcinol glycoside in the same amount with lotion containing 4-n-butylresorcinol. As a result of the above test and comparison of the feeling of use, all volunteers replied that the skin lotion containing 4-n-butylresorcinol glycoside had less irritation to the skin.

<Lotion>
Using the following formulation, a two-layer lotion was prepared by a conventional method. This product was an excellent cosmetic product that, when applied to the skin, whitens the skin and gives the skin tension and luster. Moreover, since this product contained 3-O-α-D-monoglucopyranosyl-4-n-butylresorcinol, the storage stability was also good.
(Prescription) (%)
(1) Squalane 8
(2) Tetraoleic acid polyoxyethylene sorbit 0.3
(3) Sorbit 34
(4) Pullulan 0.001
(5) Ethyl alcohol 1
(6) 3-O-α-D-glucopyranosyl-4-n-butylresorcinol prepared by the method of Example 7 0.5
(7) L-ascorbic acid-2-glucoside 2
(8) Astringent (calamine) 0.1
(9) Preservative appropriate amount (10) perfume appropriate amount (11) remaining amount of purified water

<Emulsion>
An emulsion was prepared by a conventional method using the following formulation. This product was an excellent cosmetic product that, when applied to the skin, whitens the skin and gives the skin tension and luster. Moreover, since this product contains 1-O-α-D-glucopyranosyl-4-n-butylresorcinol without generation of starch or browning, the storage stability was also good.
(Prescription) (%)
(1) Polyoxyethylene sorbitan monostearate (20E.O.) 1
(2) Polystearic acid polyoxyethylene sorbite (60E.O.) 0.5
(3) Glycerol monostearate 1
(4) Stearic acid 0.5
(5) Behenyl alcohol 0.5
(6) Squalane 8
(7) 1-O-α-D-glucopyranosyl-4-n-butylresorcinol 1 prepared by the method of Example 7
(8) Preservative 0.1
(9) Carboxyvinyl polymer 0.1
(10) Sodium hydroxide 0.05
(11) Ethyl alcohol 5
(12) Water remaining amount (13) Perfume appropriate amount

<Emulsion>
An emulsion was prepared by a conventional method using the following formulation. This product was an excellent cosmetic product that, when applied to the skin, whitens the skin and gives the skin tension and luster. Moreover, generation | occurrence | production of a starch, browning, etc. were not recognized for this product, but since 3-O- (alpha) -D-glucopyranosyl-4-n-butyl resorcinol was included, the storage stability was also favorable.
(Prescription) (%)
(1) Polyoxyethylene (20) Polyoxypropylene (2) Cetyl ether 1
(2) Methylpolysiloxane (Shin-Etsu Chemical Co., Ltd., trade name “KF96-A”) 2
(3) Liquid paraffin (medium viscosity) 3
(4) Propylene glycol 5
(5) 3-O-α-D-glucopyranosyl-4-n-butylresorcinol 1 prepared by the method of Example 7
(6) Glycerin 2
(7) Ethyl alcohol 15
(8) Carboxyvinyl polymer 0.25
(9) Hydroxypropyl cellulose 0.1
(10) Pullulan 0.05
(11) 2-Aminomethylpropanol 0.1
(12) Preservative appropriate amount (13) Purified water remaining

  The composition of the present invention can be used in various applications such as cosmetics, quasi-drugs, and external medicines. Especially, since it is excellent in the antioxidant effect and / or whitening effect, it is particularly useful as a cosmetic. Moreover, since all of the 4-n-alkylresorcinol glycosides of the present invention are highly water-soluble and have an excellent effect, they can be used for various uses, preferably as a skin whitening composition for external use.

Claims (1)

A glycoside in which a glycosyl group is α-bonded to one or both of the 1-position and 3-position hydroxyl groups in 4-alkylresorcinol, and the 4-position alkyl group is an n-methyl group or an n-ethyl group. A 4-alkylresorcinol glycoside which is an n-butyl group, an n-hexyl group, an n-heptyl group or an n-octyl group and the glycosyl group is a glucosyl group or a maltosyl group, and potassium 4-methoxysalicylate, A whitening agent for an external composition for skin having an antioxidant action, comprising one or more selected from kojic acid, arbutin and chamomile extract.

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