External composition containing purine nucleoside crystal

Abstract

[Problem] The present invention addresses the problem of providing: an external composition containing a nucleoside; a method for producing the external composition; and a cosmetic product containing the external composition. [Solution] An external composition containing a purine nucleoside crystal can be obtained by applying a dephosphorylation enzyme to a purine-containing nucleotide. In addition, when the external composition containing the purine nucleoside crystal is applied to the skin, not only gloss can be imparted to the skin but also a soft-focusing effect and a tone-up effect can also be imparted to the skin.

Description

External composition containing purine nucleoside crystals

[0001] The present invention relates to an external composition containing purine nucleoside crystals.

[0002] Guanine crystals are reflective crystals of purine bases, and are known as a substance contained in fish scales. A method for extracting this guanine from fish debris is known (Patent Document 1). In addition, due to its high reflectivity, guanine is used in cosmetic applications, particularly as a glitter material (Patent Document 2).

[0003] On the other hand, inosinic acid and guanylic acid have taste properties and are used as seasonings. Guanylic acid is composed of guanine, which is a purine base, and ribose phosphate, while inosinic acid is composed of hypoxanthine, which is a purine base, and ribose phosphate (Patent Document 3).

[0004] However, it has not been known that nucleosides such as guanosine and inosine have reflectivity, nor has it been known that they can be used as cosmetic materials that impart gloss.

[0005] Japanese Patent Application Laid-Open No. 11-130973, Japanese Patent Application Laid-Open No. 4-128211, Japanese Patent Application Laid-Open No. 2002-101846

[0006] An object of the present invention is to provide an external composition containing a purine nucleoside, a method for producing the same, and a cosmetic containing the external composition.

[0007] The inventors of the present invention conducted intensive studies to solve the above problems, and found that by allowing a dephosphorylase to act on a purine-containing nucleotide, plate-like crystals of a nucleoside having light reflectivity can be obtained, thereby completing the present invention.

[0008] In other words, the present invention relates to the following (1) to (16): (1) A topical composition comprising purine nucleoside plate-like crystals. (2) The topical composition according to (1), wherein the purine nucleoside is guanosine or inosine. (3) The topical composition according to (2), wherein the guanosine is a plate-like crystal having peaks at diffraction angles (2θ) of 20.5±0.2°, 23.7±0.2°, 25.7±0.2° and 28.2±0.2° in powder X-ray diffraction using CuKα as an X-ray source. (4) The topical composition according to (3), further having peaks at diffraction angles (2θ) of 35.5±0.2°, 39.6±0.2°, 41.6±0.2° and 47.2±0.2° in powder X-ray diffraction using CuKα as an X-ray source. (5) A method for producing the topical composition described in (1) or (2), comprising the steps of reacting a phosphatase on a purine nucleotide and obtaining guanosine or inosine. (6) A method for producing the topical composition described in (3), comprising the steps of reacting a phosphatase on guanylic acid and obtaining guanosine crystals. (7) A cosmetic product comprising the topical composition described in (1) or (2). (8) A cosmetic product comprising the topical composition described in (3). (9) A method for producing a cosmetic product, comprising the step of adding the topical composition described in (1) or (2). (10) A method for producing a cosmetic product, comprising the step of adding the topical composition described in (3). (11) A method for imparting gloss to the skin, characterized by applying the topical composition described in (1) or (2) to the skin. (12) A method for imparting gloss to the skin, characterized by applying the topical composition described in (3) to the skin. (13) A method for imparting soft focus to the skin, characterized by applying the topical composition described in (1) or (2) to the skin. (14) A method for imparting a soft focus to the skin, characterized by applying the topical composition described in (3) to the skin. (15) A method for brightening the skin tone, characterized by applying the topical composition described in (1) or (2) to the skin. (16) A method for brightening the skin tone, characterized by applying the topical composition described in (3) to the skin.

[0009] According to the present invention, a topical composition containing purine nucleoside crystals can be obtained by reacting purine nucleotides with a dephosphorylation enzyme. Furthermore, by applying this topical composition containing purine nucleoside crystals to the skin, it is possible to not only give the skin a glossy appearance but also to provide a soft-focus effect and a tone-up effect.

[0010] Comparison of powder X-ray diffraction spectra of the guanosine crystal of the present invention and reagent-grade guanosine crystal.

[0011] The topical composition of the present invention contains purine nucleoside crystals. The purine nucleoside crystals of the present invention are not limited as long as they are nucleoside crystals containing a purine base in their structure, but guanosine or inosine is preferred. Furthermore, it is desirable that the guanosine crystals or inosine crystals contained in the topical composition of the present invention be fine, plate-like crystals that are light-reflecting.

[0012] The topical composition of the present invention can be produced by reacting a purine nucleotide with a phosphatase to precipitate purine nucleoside crystals. The purine nucleotide that can be used to produce the topical composition of the present invention may be any generally available purine nucleotide, for example, guanylic acid (guanosine monophosphate) or inosinic acid (inosine monophosphate) is preferred, and a mixture of two or more purine nucleotides may also be used. When guanylic acid is reacted with a phosphatase, guanosine is obtained, and when inosinic acid is reacted with a phosphatase, inosine is obtained. Such purine nucleotides may contain other components as long as they do not inhibit the effects of the present invention.

[0013] Furthermore, the purine nucleoside crystal contained in the topical composition of the present invention is preferably guanosine, and more preferably plate-like guanosine crystals having peaks at diffraction angles (2θ) of 20.5°±0.2° preferably 0.1°, 23.7°±0.2° preferably 0.1°, 25.7°±0.2° preferably 0.1°, and 28.2°±0.2° preferably 0.1° in powder X-ray diffraction using CuKα as an X-ray source. It is even more preferably plate-like crystals having peaks at diffraction angles (2θ) of 35.5±0.2° preferably 0.1°, 39.6±0.2° preferably 0.1°, 41.6±0.2° preferably 0.1°, and 47.2±0.2° preferably 0.1°.

[0014] The crystalline form of the purine nucleoside contained in the topical composition of the present invention can be determined by microscopic observation, such as that performed by a scanning electron microscope (SEM).

[0015] Purine nucleosides such as guanosine and inosine can be quantified by HPLC. For example, a purine base standard prepared at 20 mg / mL under the following conditions can be measured to determine retention time and peak height, then each sample can be measured, and the purine base content in the sample can be calculated from its peak height. <HPLC conditions> ・Column: Develosil ODS-UG-5 (4.6 × 150 mm) (Nomura Chemical Co., Ltd.) 2 columns in tandem ・Flow rate: 0.5 mL / min ・Temperature: 50°C ・Detection wavelength: 254 nm ・Mobile phase (3 L): Potassium dihydrogen phosphate 1.02 g / 10% tetrabutylammonium hydroxide solution 9.75 mL, 85% phosphoric acid 39 mL

[0016] The nucleotides that can be used in the present invention are preferably biologically derived nucleotides obtained from biologically derived materials containing purine nucleotides. Examples of sources for such nucleotides include fish and microorganisms, with particular preference being microorganisms such as Torula yeast (Siberian Donella jadini), budding yeast (Saccharomyces cerevisiae), and Bacillus subtilis. Microbial nucleotides can be obtained by culturing the microorganisms according to general methods and then extracting them under appropriate conditions, or by using commonly available yeast extracts or Bacillus subtilis extracts containing purine nucleotides.

[0017] Furthermore, the dephosphorylating enzyme that can be used in the production of the topical composition of the present invention is not limited as long as it is an enzyme that has the activity to hydrolyze phosphate monoesters, and in particular, an enzyme having nucleotidase activity (EC3.1.3.5 or EC3.1.3.6) that uses nucleotides as substrates can be preferably used.

[0018] In the process of producing the cosmetic material composition of the present invention, the amount of dephosphorylating enzyme added and the conditions under which the dephosphorylating enzyme is applied are not limited as long as conditions are available for producing nucleosides from nucleotides, and can be set as appropriate. Examples include pH 3 to 5 and 30°C to 50°C. The enzyme reaction time can be set as appropriate according to the amount of nucleosides produced.

[0019] The process may include steps such as reacting nucleotides containing purine bases with a dephosphorylation enzyme, precipitating crystals, and removing soluble components by filtration, or reacting nucleotides containing purine bases with a nucleosidase, followed by resin purification, or further drying after crystal precipitation. The method for precipitating nucleotide crystals containing purine bases is not particularly limited as long as nucleotide crystals containing purine bases are obtained, and general precipitation methods can be employed. For example, a solution or suspension containing nucleotides may be heated and dissolved, then cooled, or a poor solvent method, evaporation method, or pH-controlled precipitation method may be employed.

[0020] The external composition obtained by the method described above can be used as is, or combined with thickeners, pH adjusters, fragrances, bases for external preparations, excipients, etc., commonly used in cosmetics, to make cosmetics. Examples include cosmetics such as serums, foundations, mascaras, nail polishes, hair glitters, and lipsticks. When manufacturing cosmetics using the external composition of the present invention, it may be added at the same time as other ingredients commonly used in cosmetic ingredients, or at different times.

[0021] The topical composition of the present invention contains fine plate-like crystals of purine nucleoside, which reflect light, and therefore can impart a glossy effect to the skin when applied. The glossy effect can be visually determined after applying the topical composition of the present invention to the skin. The glossy effect can be confirmed by using the cosmetic material composition of the present invention as is or by incorporating it into cosmetics.

[0022] Furthermore, because the external composition of the present invention reflects light due to the plate-like crystals of fine purine nucleosides, it can impart a soft-focus effect to the skin when applied. Here, the soft-focus effect in the present invention refers to the effect of making it difficult to see what is underneath the powder by the particles applied to the skin reflecting light, and is known as the effect of blurring unevenness such as fine lines and pores. The soft-focus effect can be confirmed by visual inspection after applying the cosmetic material composition of the present invention to the skin as is or after incorporating it into cosmetics.

[0023] Furthermore, because the topical composition of the present invention reflects light due to the plate-like crystals of fine purine nucleosides, it can provide a tone-up effect to the skin when applied. Here, the tone-up effect in the present invention refers to the effect of brightening the overall skin tone by covering uneven skin color and making the color uniform through the reflection of light by the particles applied to the skin. The tone-up effect can be confirmed by visual inspection after applying the cosmetic material composition of the present invention to the skin, either as is or after incorporating it into cosmetics.

[0024] The present invention will be described in detail below with reference to examples. However, the present invention is not limited to the following embodiments. The guanosine and inosine content was measured using the method described above.

[0025] <Example 1> A 10% solution of commercially available reagent-grade guanylic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) was adjusted to pH 3, and 1.5% of commercially available dephosphorylation enzyme was added to the guanylic acid. The reaction was carried out at 50°C for 3 hours. The reaction solution was heated to 90°C and then rapidly cooled to precipitate crystals. The suspension was filtered by suction and the precipitate was collected. The collected precipitate was allowed to stand at 25°C for 10 hours to obtain a dried product. The guanosine content in this dried product was 95%. The obtained crystals were crushed in a mortar and pestle and observed by SEM, revealing plate-like crystals as shown in Figure 1.

[0026] <Example 2> The same procedure was carried out using commercially available reagent-grade inosinic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) instead of guanylic acid as in Example 1, to obtain a dried product. The inosine content in this dried product was 94%.

[0027] <Example 3> The same procedure was carried out using "Ribotide" (manufactured by Mitsubishi Corporation Life Sciences Co., Ltd.), a nucleic acid seasoning which is a mixture of guanylic acid and inosinic acid, instead of guanylic acid as in Example 1, to obtain a dried product. The guanosine content in this dried product was 48%, and the inosine content was 45%.

[0028] <Reflectivity Confirmation Test - Sensory Evaluation> The dried products obtained in Examples 1 to 3 were applied to the skin and their reflectivity was confirmed. In addition, as samples that had not been treated with dephosphorylation enzymes, Comparative Example 1 (guanylic acid), Comparative Example 2 (inosinic acid), and Comparative Example 3 (nucleic acid seasoning containing guanylic acid and inosinic acid) were applied to the skin and their light reflectivity was confirmed. Reflectivity was confirmed visually, and a maximum of 5 points was given for high reflectivity, and a minimum of 1 point for low reflectivity.

[0029] <Reflectivity Confirmation Test - Sensory Evaluation Results> The results are shown in Table 1. From these results, it was clear that the test groups of samples treated with dephosphorylation enzyme (Examples 1-3) all showed high reflectivity. On the other hand, the test groups of samples not treated with dephosphorylation enzyme (Comparative Examples 1-3) showed poor reflectivity. Furthermore, the results of this test showed that by increasing the light reflectivity of the skin, it was possible to impart gloss to the skin, and that it had excellent soft-focus and tone-up effects. From these findings, it was considered that not only Example 1 (guanosine) but also Example 2 (inosine) were plate-like crystals. Furthermore, from the results of Example 3, it was considered that a mixture of guanylic acid and inosinic acid treated with dephosphorylation enzyme also formed plate-like crystals.

[0030]

[0031] <Powder X-ray Diffraction> The dried material obtained in Example 1 and reagent-grade guanosine powder (Comparative Example 4; manufactured by Tokyo Chemical Industry Co., Ltd.) were subjected to powder X-ray diffraction under the measurement conditions described later to analyze their crystal structures.

[0032] <Measurement Conditions for Powder X-ray Diffraction> Measurement device: Aeris Research edition (Malvern Panalogical) X-ray output setting: 15 mA, 40 kV Goniometer radius [mm]: 145.00 Scan axis: Gonio Start position [°2Th.]: 5.0407 End position [°2Th.]: 69.9707 Step size [°2Th.]: 0.0430 Scan step time [s]: 96.3900 Scan type: Continuous PSD mode Scan PSD distance [°2Th.]: 5.54 Offset [°2Th.]: 0.0000 Divergent slit (DS) type Fixed divergent slit (DS) size [°]: 0.1146 Sample width [mm]: 10.00 Measurement temperature [°C] 25.00 Target: Cu Intended Wavelength Type: K-α1 Distance between focus and DS [mm]: 95.00 Incident monochromator: None

[0033] <Measurement Results of Powder X-ray Diffraction> The results of powder X-ray diffraction are shown in Chart 1 for Example 1 and Chart 2 for Comparative Example 4 in Figure 2. From these results, it became clear that the guanosine crystal of the present invention has a different diffraction pattern from commercially available reagent-grade guanosine.

[0034] <Reflectivity Confirmation Test - Brightness> 0.2 g of the dried material from Example 1 or 0.2 g of the sample from Comparative Example 4 (reagent-grade guanosine) was mixed with 2 g of commercially available acrylic resin and uniformly applied to a black substrate. After curing the resin by irradiating it with UV light for 5 minutes, the brightness (L value) was measured using a spectrophotometer CM-5 (manufactured by Konica Minolta Japan, Inc.). A sample in the same procedure without adding anything to the acrylic resin was used as control example 1.

[0035] <Reflectivity Confirmation Test - Brightness Results> The results are shown in Table 2. From these results, it was shown that Example 1, which was treated with dephosphorylation enzyme, had higher brightness and was brighter than Comparative Example 4.

[0036]

[0037] The results above demonstrate that the topical composition of the present invention can be used as a cosmetic. Furthermore, it was shown that when applied to the skin, it gives the skin a glossy appearance and provides a soft-focus effect and a tone-up effect.

Claims

1. A topical composition containing purine nucleoside plate-like crystals.

2. The topical composition according to claim 1, wherein the purine nucleoside is guanosine or inosine.

3. The topical composition according to claim 2, wherein the guanosine is a plate-like crystal having peaks at diffraction angles (2θ) of 20.5±0.2°, 23.7±0.2°, 25.7±0.2°, and 28.2±0.2° in powder X-ray diffraction using CuKα as an X-ray source.

4. The topical composition according to claim 3, wherein, in powder X-ray diffraction using CuKα as an X-ray source, the composition further has peaks at diffraction angles (2θ) of 35.5±0.2°, 39.6±0.2°, 41.6±0.2°, and 47.2±0.2°.

5. A method for producing the topical composition according to claim 1 or 2, comprising the steps of reacting a purine nucleotide with a dephosphorylating enzyme and obtaining guanosine or inosine.

6. A method for producing the topical composition according to claim 3, comprising the steps of reacting guanylic acid with a dephosphorylating enzyme and obtaining guanosine crystals.

7. A cosmetic comprising the external composition according to claim 1 or 2.

8. A cosmetic comprising the external composition described in claim 3.

9. A method for producing a cosmetic product, comprising the step of adding the topical composition described in claim 1 or 2.

10. A method for producing cosmetics, comprising the step of adding the topical composition described in claim 3.

11. A method for imparting gloss to skin, characterized by applying the topical composition described in claim 1 or 2 to the skin.

12. A method for imparting gloss to skin, characterized by applying the topical composition described in claim 3 to the skin.

13. A method for imparting a soft-focus effect to skin, characterized by applying the topical composition described in claim 1 or 2 to the skin.

14. A method for imparting a soft-focus effect to the skin, characterized by applying the topical composition described in claim 3 to the skin.

15. A method for brightening skin tone, characterized by applying the topical composition described in claim 1 or 2 to the skin.

16. A method for brightening skin tone, characterized by applying the topical composition described in claim 3 to the skin.

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