JP5226982B2 - A topical skin preparation containing an extract of buckwheat - Google Patents

Description

  The present invention relates to a skin external preparation containing a plant extract.

  Conventionally, whitening cosmetics have been widely used for the purpose of obtaining cosmetic effects such as prevention of skin darkness, spots and freckles, but these are important enzymes in the melanin production pathway that causes pigmentation. Many try to inhibit tyrosinase. Such whitening cosmetics mainly contain ascorbic acid, glutathione, kojic acid, colloidal sulfur and the like. However, ascorbic acid and glutathione are susceptible to oxidation, kojic acid still has safety concerns, and colloidal sulfur has the disadvantages of producing off-flavors and precipitation.

  For this reason, recently, a search for a whitening agent that is safe in nature has been conducted, and many plant extracts exhibiting tyrosinase inhibitory activity have been proposed. For example, tartary buckwheat (Fagopyrum tataricum (L.) Garten.), Which is cultivated and eaten in Yunnan Province and Nepal, China, has an extract of cis-umberic acid. It has been proposed that it has the ability to inhibit tyrosinase and is effective for beauty (see Patent Documents 1 to 8).

  The rutin content of tartary buckwheat is about 100 times that of buckwheat and is very high. Rutin is a kind of flavonoid, is a functional ingredient that has a capillary strengthening action and is effective for diseases such as hypertension. However, tartary buckwheat seeds are also rich in rutin-degrading enzymes, and it is said that quercetin, a bitter component, is rapidly decomposed by addition to flour. Tartary buckwheat has good fertility and high yield, so it has been tried to cross with ordinary buckwheat, but it has not been successful because of the different species.

  On the other hand, buckwheat (scientific name Fagopyrum esculetum) is an annual herb belonging to the family Tedaceae, similar to tartary buckwheat, but differs from tartary buckwheat as a species. About 30% of buckwheat berries are buckwheat, and about 7,000 tons are discharged as agricultural waste per year, which is currently attracting attention as an unused resource. For example, attempts have been made to find biological effects and use them as drugs. For example, it has been proposed for amylolytic enzyme inhibitory ability, anti-influenza action, and Maillard reaction inhibition (see Patent Documents 9 to 11).

  In addition, as for buckwheat, its use as a deodorant has been proposed using its physicochemical properties (see Patent Document 12). Furthermore, lipid peroxide inhibitors, cholesterol-lowering agents, neutral fat-lowering agents, or hyperlipidemia-improving agents, which contain as an active ingredient an extract obtained by extracting buckwheat with water or an organic solvent alone or a mixture thereof. Has also been proposed (see Patent Document 13). And the collagenase activity inhibitor which consists of an aqueous protein extract of buckwheat has also been proposed (refer patent document 14). In addition, it has been proposed to prepare a buckwheat extract by heating extraction of buckwheat with ethanol (see Patent Document 15).

  On the other hand, elastin, which has the function of maintaining skin elasticity and elasticity, is denatured and destroyed by the action of elastase that is overexpressed by UV exposure and aging, and as a result, the elasticity of the skin decreases, leading to skin aging. It has been. For this reason, many studies have been made to obtain substances exhibiting elastase inhibitory activity in plant extracts (see Patent Documents 16 to 20).

Japanese Examined Patent Publication No. 62-045202 Japanese Patent Publication No. 48-1505 Japanese Patent Publication No. 48-29139 Japanese Patent Laid-Open No. 02-45408 Japanese Patent Laid-Open No. 01-319473 Japanese Patent No. 0234595 Japanese Examined Patent Publication No. 56-18869 Japanese Patent Laid-Open No. 60-16906 Japanese Patent Laid-Open No. 2005-220110 Japanese Patent Application Laid-Open No. 2004-2361 JP 2006-256977 A JP 2003-305112 A JP-A-10-218786 Japanese National Patent Publication No. 8-507506 JP 2002-272396 A Japanese Patent Application Laid-Open No. 10-017460 Japanese Unexamined Patent Publication No. 2000-026263 JP-A-11-263720 Japanese Patent Laid-Open No. 11-246388 JP 11-335235 A

  However, in the above-mentioned Patent Document 6, since cis-umbellic acid is contained in the grain extract, tartary buckwheat described as having a tyrosinase inhibiting ability and being effective for beauty is often a raw material. I couldn't say that I was satisfied with the availability and price. For example, tartary buckwheat has a limited amount of cultivated land in Japan compared to ordinary buckwheat, domestic harvest is about 3t, and about 600t is imported. It does not enter.

  In addition, in Patent Documents 13 to 15 described above, it is described that there are some physiologically active functions with respect to the extract of buckwheat, but no report on the whitening effect due to the ability to inhibit tyrosinase has been made yet. In other words, until now, it is unknown whether buckwheat, an industrial waste obtained from buckwheat, a plant that is abundantly available at low cost, contains an active ingredient that exhibits a whitening effect due to its ability to inhibit tyrosinase. I must say.

  Further, in Patent Documents 16 to 20 described above, there are described research results obtained by using a plant extract for a substance exhibiting elastase inhibitory activity, but there is no particular description regarding buckwheat. That is, to date, it is unknown whether buckwheat, an industrial waste obtained from buckwheat, an abundant and inexpensively available plant, contains an active ingredient that exhibits an anti-aging effect due to its elastase inhibitory ability. I must say that there was.

  The present invention has been made in view of the above circumstances, and an object thereof is to search for a plant extract exhibiting a whitening action and a skin beautifying action and an active ingredient thereof.

  ADVANTAGE OF THE INVENTION According to this invention, the skin external preparation containing the extract of buckwheat is provided. Since the extract of buckwheat contained in the external preparation for skin of the present invention has a tyrosinase inhibiting ability and an elastase inhibiting ability, the external skin preparation of the present invention itself also has a tyrosinase inhibiting ability and an elastase inhibiting ability. Therefore, the skin external preparation of this invention shows both the whitening effect | action by tyrosinase inhibitory ability, and the anti-aging effect | action by elastase inhibitory ability.

  In the skin external preparation of the present invention, the above extract of buckwheat contains orientin, isoorientin, vitexin, isovitexin and rutin derived from buckwheat. Since these five kinds of components all have tyrosinase inhibitory ability and elastase inhibitory ability, the external preparation for skin of the present invention itself also has tyrosinase inhibitory ability and elastase inhibitory ability. However, as will be described later, the above extract of buckwheat has remarkably superior tyrosinase inhibitory ability and elastase inhibitory ability than a mixture obtained by simply mixing these five kinds of components.

  Moreover, according to this invention, the tyrosinase inhibitor containing the extract of buckwheat is provided. Since the extract of buckwheat contained in the tyrosinase inhibitor of the present invention has the ability to inhibit tyrosinase, the tyrosinase inhibitor itself of the present invention also has the ability to inhibit tyrosinase.

  Moreover, according to this invention, the tyrosinase inhibitor containing orientin or isoorientin is provided. Since either orientin or isoorientin contained in the tyrosinase inhibitor of the present invention has a tyrosinase inhibitory ability, the tyrosinase inhibitor itself of the present invention also has a tyrosinase inhibitory ability.

  Moreover, according to this invention, the elastase inhibitor containing the extract of buckwheat is provided. Since the extract of buckwheat contained in the tyrosinase inhibitor of the present invention has an elastase inhibitory ability, the elastase inhibitor itself of the present invention also has an elastase inhibitory ability.

  Moreover, according to this invention, the composition for cosmetics containing the extract of buckwheat is provided. Since the extract of buckwheat contained in the cosmetic composition of the present invention has tyrosinase inhibitory ability and elastase inhibitory ability, the cosmetic composition of the present invention itself also has tyrosinase inhibitory ability and elastase inhibitory ability. Therefore, the cosmetic composition of the present invention exhibits both a whitening effect due to tyrosinase inhibiting ability and an anti-aging effect due to elastase inhibiting ability.

  Moreover, according to the present invention, a method for producing a cosmetic composition from buckwheat, the step of depositing buckwheat with water-containing ethanol for a period of 1 day to 4 days under a temperature condition of 4 ° C. to 30 ° C., Producing a composition comprising orientin, isoorientin, vitexin, isovitexin and rutin from the extract obtained as a result of the deposition. In the production method of the present invention, buckwheat is deposited with water-containing ethanol under a temperature condition of 4 ° C. or more and 30 ° C. or less for a period of 1 day or more and 4 days or less. Can be extracted. Therefore, in the production method of the present invention, a cosmetic composition excellent in tyrosinase inhibiting ability and elastase inhibiting ability can be produced efficiently.

  According to the present invention, there is also provided a method for producing a cosmetic composition from buckwheat, wherein the buckwheat is heated with hydrous ethanol at a temperature of 40 ° C. or higher and a boiling temperature or lower for a period of 0.5 hours to 2 hours. And a step of producing a composition containing orientin, isoorientin, vitexin, isovitexin and rutin from the extract obtained as a result of heating. In the production method of the present invention, buckwheat is heated with water-containing ethanol at a temperature of 40 ° C. or higher and a boiling temperature or lower for a period of 0.5 hours or more and 2 hours or less, so that orientin, isoorientin, vitexin, isovitexin and rutin are obtained from buckwheat. It can be extracted efficiently. Therefore, in the production method of the present invention, a cosmetic composition excellent in tyrosinase inhibiting ability and elastase inhibiting ability can be produced efficiently.

  According to the present invention, the extract of buckwheat having tyrosinase inhibitory ability and elastase inhibitory ability is used, and therefore, a skin external preparation and cosmetic composition exhibiting both a whitening action by tyrosinase inhibitory ability and an anti-aging action by elastase inhibitory ability Is obtained.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings.

<Overview>
This embodiment is related with the skin external preparation containing the extract of buckwheat. More specifically, this embodiment relates to a buckwheat extract containing orientin, isoorientin, vitexin, isovitexin and rutin as main active ingredients. Furthermore, the present embodiment is based on the discovery that buckwheat extract inhibits the action of tyrosinase in melanocytes and suppresses melanin production, such as whitening effect on skin after ultraviolet irradiation, spots due to sunburn, buckwheat, etc. The present invention relates to a skin external preparation effective for prevention or treatment. Furthermore, the present embodiment is based on the discovery that the buckwheat extract inhibits the action of elastase which has the function of degrading elastin that gives elasticity to the skin, and has an anti-aging action that keeps the skin firm and elastic. It relates to a topical skin preparation.

  Here, buckwheat (scientific name Fagopyrum esculetum) is a first-year herb belonging to the family Tadeidae, and has long been a crop that cannot be separated from Japanese life, including prevention and improvement of lifestyle-related diseases and customs such as “New Year's buckwheat”. Has been cultivated as. In addition, buckwheat nuts are removed from the shell (rounded), and the endosperm portion of the seed is floured (buckwheat flour) for edible use.

  Buckwheat (Sobagara, buckwheat husk) refers to the shell that remains after harvesting buckwheat, drying it in the sun for several days, and removing the buckwheat. In addition, when refining buckwheat flour, the buckwheat is pulled together with the fruit to give a flavor. As for the use of buckwheat, it is often used as the contents of bedding pillows due to its physical characteristics.

  To reiterate, the external preparation for skin containing the plant extract of this embodiment (the extract of buckwheat) suppresses the production of melanin pigments that cause pigmentation, stains and freckles after sunburn due to the ability to inhibit tyrosinase, and whitening In addition to having an effect on the beautiful skin, the elastase inhibitory ability can keep the skin firm and elastic and prevent skin aging. Thus, the skin external preparation of this embodiment is a skin external preparation which mix | blends the extract of buckwheat which is a food waste. Therefore, the skin external preparation of this embodiment can be used as a cosmetic product having an anti-aging effect obtained using buckwheat, which is an abundant and inexpensively available raw material.

<Extraction method from buckwheat>
The extraction method of the buckwheat extract used in the present embodiment is not particularly limited. For example, from the viewpoint of improving the extraction efficiency and improving the quality of the extract, the buckwheat is immersed in ethanol at room temperature for half a day or more, or heated. It can be obtained by heating and stirring for about 1 hour and filtering, and then concentrating and lyophilizing the filtrate.

  In addition, the extraction conditions for the buckwheat extract used in the present embodiment are not particularly limited, but from the viewpoint of improving extraction efficiency and improving the quality of the extract, it is usually in the range of 1 to 2 L per 100 g of buckwheat. Water or an organic solvent such as cold or hot ethanol, methanol, propanol, isopropanol, propylene glycol, butylene glycol, glycerin, butanol, acetone, methyl ethyl ketone, and dioxane can be preferably used. Among these, from the viewpoint of further improving the extraction efficiency and improving the quality of the extract, it is preferable to use a mixed solvent (hydrous organic solvent) of these organic solvents and water, and further comprises ethanol, isopropanol, butylene glycol, and acetone. It is more preferable to use a mixed solvent of at least one organic solvent selected from the group and water, and it is most preferable to use a mixed solvent of ethanol and water.

  When a mixed solvent of ethanol and water is used, the ethanol composition is in the range of 10% (V / V) to 90% (V / V) from the viewpoint of improving extraction efficiency and improving the quality of the extract. The composition of ethanol is preferably 50% (V / V) or more and 70% (V / V) or less. In the case of using an organic solvent other than ethanol, the content of the same organic solvent (preferably 10% (V / V) or more and 90% (V / V) or less, from the viewpoint of improving extraction efficiency) A mixed solvent having 50% (V / V) or more and 70% (V / V) or less) is preferable.

  On the other hand, the combination of extraction temperature and extraction time of the buckwheat extract used in the present embodiment is immersed for 1 day or more and 4 days or less under a temperature condition of 4 ° C. or higher and 30 ° C. or lower from the viewpoint of improving extraction efficiency and extract quality. Then, it is preferable to deposit the solid content of buckwheat and extract the supernatant.

  Alternatively, as another combination of the extraction temperature and extraction time of the buckwheat extract used in the present embodiment, from the viewpoint of improving the extraction efficiency and improving the quality of the extract, it is 0.5 under a temperature condition of about 40 ° C. or higher and the boiling temperature or lower. It is preferable to heat for a period of not less than 2 hours but not more than 2 hours, and most preferable is to heat and reflux at the boiling temperature for 1 hour.

<Activity of buckwheat extract>
The action and effect of the extract will be described in detail in the examples described later. However, as described earlier from the conclusion, the buckwheat extract obtained from buckwheat by the above extraction method is obtained by high performance liquid chromatography (HPLC). When analyzed, five components are mainly detected (FIG. 1). When these five components are isolated using various separation methods and subjected to structural analysis, the flavonoids orientin, isoorientin, vitexin, isovitexin and rutin are obtained (FIG. 3).

  Here, tyrosinase inhibitory activity for some of these flavonoids (vitexin, isovitexin and rutin) and elastase inhibitory activity for all flavonoids (orientin, isoorientin, vitexin, isovitexin and rutin) Although reported by a patent contributor, etc., no tyrosinase inhibitory activity has been reported for orientin and isoorientin.

  The tyrosinase inhibitory activity for orientin and isoorientin will be described in detail in the examples below. However, as described earlier from the conclusion, the present inventors have stated that orientin and isoorientin have tyrosinase inhibitory activity. It has been discovered for the first time in an experiment described in Examples described later.

  On the other hand, the comparison of the action and effect of the buckwheat extract and the mere mixture of the above five components will be described in detail in the examples described later. A composition composed of only five flavonoids contained was separately prepared, and the enzyme inhibitory activities (tyrosinase inhibitory activity and elastase inhibitory activity) of this mixed composition and the extract of buckwheat were compared.

  As a result, the present inventors found that the extract of buckwheat extract showed superior activity in any of the tests (tyrosinase inhibitory activity test and elastase inhibitory activity test) (FIG. 6). It was concluded that is useful compared to the above mixed composition in terms of tyrosinase inhibitory activity and elastase inhibitory activity.

<Effect>
Hereinafter, the operational effects of the present embodiment will be described in detail.
The extract of buckwheat (the extract of buckwheat) contained in the external preparation for skin (cosmetic composition) of this embodiment has a tyrosinase inhibitory ability and an elastase inhibitory ability. Therefore, the skin external preparation (cosmetic composition) of the present embodiment exhibits both a whitening effect due to the tyrosinase inhibitory ability and an anti-aging effect due to the elastase inhibitory ability.

  In the external preparation for skin (cosmetic composition) of the present embodiment, the above extract of buckwheat (buckthorn extract) contains buckwheat-derived orientin, isoorientin, vitexin, isovitexin and rutin. Since these five kinds of components all have tyrosinase inhibitory ability and elastase inhibitory ability, the external preparation for skin (cosmetic composition) of this embodiment has a whitening action by tyrosinase inhibitory ability and an anti-aging action by elastase inhibitory ability. Both are shown.

  However, as will be described later in the Examples, the above extract of buckwheat (the extract of buckwheat) has a significantly superior tyrosinase inhibitory ability and elastase inhibitory ability than a mixture obtained by simply mixing these five components. (FIG. 6).

  Here, in the external preparation for skin (cosmetic composition) of this embodiment, buckwheat is extracted with a water-containing organic solvent, and therefore, five kinds of active ingredients of orientin, isoorientin, vitexin, isovitexin and rutin from buckwheat are included. Since the extraction efficiency and the quality of the extract are excellent, it is possible to obtain a skin external preparation (cosmetic composition) excellent in both the whitening action by the tyrosinase inhibitory ability and the anti-aging action by the elastase inhibitory ability.

  Moreover, when using the water-containing organic solvent containing at least one organic solvent selected from the group consisting of ethanol, isopropanol, butylene glycol, and acetone as the water-containing organic solvent, orientin, isoorientin, vitexin, Since the extraction efficiency and quality of the extract of five kinds of active ingredients of isovitexin and rutin are improved, the skin external preparation (cosmetic composition) is further improved by the whitening action by the tyrosinase inhibitory ability and the anti-aging action by the elastase inhibitory ability. ) Is obtained.

  Moreover, when using a water-containing organic solvent having an organic solvent content of 10% (v / v) or more and 90% (v / v) or less as the water-containing organic solvent, orientin, isoorientin, vitexin, Since the extraction efficiency and quality of the extract of five kinds of active ingredients of isovitexin and rutin are improved, the skin external preparation (cosmetic composition) is further improved by the whitening action by the tyrosinase inhibitory ability and the anti-aging action by the elastase inhibitory ability. ) Is obtained.

  Further, the above extract of buckwheat (the extract of buckwheat) can also be used as a tyrosinase inhibitor. The extract of buckwheat contained in this tyrosinase inhibitor (the extract of buckwheat) has a tyrosinase inhibitory ability, and can therefore be suitably used as a tyrosinase inhibitor.

  Further, orientin or isoorientin, which is a component contained in the above extract of buckwheat, can be used as a single component or as a tyrosinase inhibitor. Since either orientin or isoorientin contained in this tyrosinase inhibitor has tyrosinase inhibitory ability, it can be suitably used as a tyrosinase inhibitor.

  Also, the above extract of buckwheat (the extract of buckwheat) can be used as an elastase inhibitor. The extract of buckwheat contained in the elastase inhibitor of the present embodiment has an elastase inhibitory ability and can therefore be suitably used as an elastase inhibitor.

  In addition, when the above extraction method is used, when producing a cosmetic composition (external skin preparation) from buckwheat, the buckwheat is treated with water-containing ethanol at a temperature of 4 ° C. to 30 ° C. for a period of 1 day to 4 days. Since it is deposited, orientin, isoorientin, vitexin, isovitexin and rutin can be efficiently extracted from buckwheat. Therefore, in this production method, a cosmetic composition excellent in tyrosinase inhibiting ability and elastase inhibiting ability can be produced efficiently.

  On the other hand, when the above-described another extraction method is used, when producing a cosmetic composition from buckwheat, the buckwheat is heated with water-containing ethanol for a period of 0.5 hours to 2 hours at a temperature of 40 ° C. Therefore, orientin, isoorientin, vitexin, isovitexin and rutin can be efficiently extracted from buckwheat. Therefore, in this production method, a cosmetic composition excellent in tyrosinase inhibiting ability and elastase inhibiting ability can be produced efficiently.

  As mentioned above, although embodiment of this invention was described with reference to drawings, these are the illustrations of this invention, Various structures other than the above are also employable.

  For example, in the above-described embodiment, the extract contained in the external preparation for skin is a buckwheat extract, but this is not intended to exclude the extract of buckwheat berries, but some buckwheat fruit fragments inside the buckwheat etc. As a result, the extract of buckwheat berries may be partially mixed with the extract of buckwheat. In such a case, the active ingredient contained in the extract of buckwheat and the active ingredient contained in the extract of buckwheat act additively or synergistically, and the whitening action by the tyrosinase inhibitory ability, This is because an anti-aging effect due to the elastase inhibitory ability and a more excellent external preparation for skin can be obtained.

  Or you may extract an extract as it is from what is called buckwheat grain which contains both buckwheat and buckwheat seeds as an extract contained in a skin external preparation. Even in such a case, the whitening action by the tyrosinase inhibitory ability of the active ingredient contained in the extract of buckwheat and the anti-aging action by the elastase inhibitory ability, and various physiology of the active ingredient contained in the extract of buckwheat This is because an even better skin external preparation can be obtained.

  Of course, the types and compositions of active ingredients differ between buckwheat and buckwheat, and it is assumed that buckwheat has a larger surface area than the weight and a greater content of active ingredients per unit weight. In particular, buckwheat, which is industrial waste, is cheaper than buckwheat berries consumed as food, and can be easily obtained in large quantities, so the buckwheat extract is extracted only from buckwheat. Is preferred.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further, this invention is not limited to these.

<Background>
The present inventors have so far screened about 120 kinds of herbal medicines and herbs by a tyrosinase and elastase inhibition test. The tyrosinase inhibition test aims at a whitening effect, and the elastase inhibition test aims at a skin beautifying effect.

  As a result, the present inventors found tyrosinase inhibitory activity and elastase inhibitory activity particularly strong against buckwheat, compared with other crude drugs and herbs.

  Therefore, the present inventors conducted a tyrosinase inhibitory active ingredient search for buckwheat using column chromatography, large-scale preparative HPLC, and the like. The present inventors applied EtOH extract to LH-20 column chromatography and found that tyrosinase inhibitory activity was observed in the 50% MeOH elution fraction. Therefore, HPLC analysis was performed under the following HPLC conditions, and five major peaks were observed. I found it. FIG. 1 is an HPLC chromatogram of an extract extracted from buckwheat. The present inventors tentatively named these five major peaks as peaks 1 to 5, respectively, and performed separation and identification.

* HPLC condition column: TSK gel ODS-80TM (4.0φ × 150mm)
Mobile phase: 1% acetic acid-MeOH (65:35) → (40:60)
20 minutes linear gradient 1% acetic acid-MeOH (40:60) 5 minutes flow rate: 1.0 ml / min
Temperature: 40 ° C
Detection: Photodiode array (display is 350 nm)

<Examination of extraction efficiency>
Since the extract yield was as low as 1 to 1.5% in the above initial EtOH extraction, the present inventors examined a new extraction method aiming at improving the yield of peaks 1 to 5. When the peak areas were compared by HPLC analysis, the extraction method with 50% EtOH was the most efficient. In FIG. 2, the result of having examined the extraction method and extraction efficiency from buckwheat is shown as a graph. Hereinafter, 50% EtOH (cold treatment) extract was used.

  As shown in Production Example 5 to be described later, the present inventors fractionated the components of peaks 1 to 5 thus obtained based on a fraction chart of an extract extracted from buckwheat. Then, the present inventors determined that the components of these peaks 1 to 5 are shown in FIG. 3 based on the results of the NMR analysis shown in FIG. 1 and the NMR structure analysis (data not shown) described later in Production Example 5. It confirmed that it was each component shown in. FIG. 3 shows the structural formulas of peaks 1 to 5 and tyrosinase inhibitory activity. The tyrosinase inhibitory activity will be described later in detail.

<Tyrosinase inhibitory activity test>
Melanin pigments that cause stains and buckwheat are produced by the action of the enzyme “tyrosinase” activated by UV irradiation. Therefore, if this tyrosinase is inhibited, melanin fixation can be suppressed, and a whitening effect can be expected. Therefore, the present inventors conducted a tyrosinase inhibitory activity test in order to evaluate this whitening effect. The results of the tyrosinase inhibitory activity test will be described later in detail.

<Elastase inhibitory activity test>
The skin is made up of two fibers: “elastin” that gives elasticity and “collagen” that keeps the skin soft. When the dermis is exposed to ultraviolet rays, it secretes a large amount of the enzyme “elastase” and cuts off elastin. Therefore, by inhibiting this elastase, an anti-aging effect that keeps the skin firm and elastic can be expected. Therefore, the present inventors conducted an elastase inhibitory activity test in order to evaluate this anti-aging effect. The results of the elastase inhibitory activity test will be described in detail later.

<Production Example 1> (Production example of buckwheat extract by ethanol immersion)
The present inventors immersed 100 g of buckwheat in 1000 ml of ethanol at room temperature for about half a day and filtered to obtain an extract. The present inventors further immersed the residue in 1000 ml of ethanol at room temperature for about 3 days, filtered to obtain an extract, and combined with the previous extract under reduced pressure. The present inventors further dried the obtained residue under reduced pressure at room temperature to obtain 1.54 g as a buckwheat extract (yield 1.5%).

<Production Example 2> (Production Example of Buckwheat Extract by 50% Ethanol Cooling)
The present inventors immersed 100 g of buckwheat in 1000 ml of 50% ethanol at room temperature for about one day, filtered under reduced pressure to obtain an extract, and concentrated and lyophilized to obtain 2.87 g of extract (yield 2. 9%).

<Production Example 3> (Production Example of Buckwheat Extract by 70% Ethanol Cooling)
The present inventors soaked 10 g of buckwheat in 200 ml of 70% ethanol at room temperature for about 1 day, filtered by suction to obtain an extract, concentrated and freeze-dried to obtain 179 mg of extract (yield 1.8%) ).

<Production example 4> (Production example of buckwheat extract by heating with 50% ethanol)
The present inventors added 100 g of buckwheat pulverized product to 1000 ml of 50% ethanol heated to 70 ° C., and stirred while heating for about 1 hour, and filtered under reduced pressure while hot to obtain an extract and concentrated under reduced pressure. The present inventors freeze-dried the obtained residue to obtain 3.23 g of a buckwheat extract (yield 3.2%).

<Production Example 5>
The extract obtained in Production Example 2 was adsorbed on DIAION HP-20 column chromatography (column: 6 cmφ × 36 cm), and the fractions eluted with 60% methanol were collected, and separated into Sephadex LH-20 column chromatography (column: 3.5 cmφ). X40 cm) and the portion eluted with 50% methanol was collected. Subsequently, fractionation was performed by large-scale preparative HPLC (column: Luna 5 μC18, 21.2 mmφ × 250 mm, Phenomenex; solvent: 1% acetic acid / methanol = 70: 30), and the peak 1-2 fractions and peak 3 shown in FIG. Fraction (68 mg), peak 4 fraction (137 mg) and peak 5 fraction (41 mg) were obtained. Further, the fractions of peaks 1 and 2 were fractionated with 1% acetic acid / methanol = 75: 25 to obtain peak 1 (57 mg) and peak 2 (45 mg).

  The present inventors have confirmed that these peaks 1 are orientin, peak 2 is isoorientin, peak 3 is vitexin, peak 4 is isovitexin and peak 5 is rutin. It was identified by comparing the data (data not show) with the values shown in the following references.

Reference 1: M. Watanabe et.al., J. Agric. Food Chem., 45, 1039-1044 (1997).
Reference 2: T. Ozawa et.al., Biosci. Biotechnol. Biochem., 67 (2), 410-414 (2003).
Reference 3: T. Sabudak et.al., Trakya Univ. J. Sci., 6 (2) 88-92 (2005).

<Test Example 1> (Tyrosinase inhibition test)
a) Test method The inventors conducted the test as follows. That is, a mixed solution of 10 μl of a sample solution such as buckwheat extract (50% ethanol extract), 50 μl of 50 mM phosphate buffer, 20 μl of distilled water and 20 μl of mushroom-derived tyrosinase solution was incubated at 37 ° C. for 5 minutes. Subsequently, 100 μl of L-tyrosine was added and the reaction was carried out at 37 ° C., and the amount of produced dopaquinone was measured over time with a microplate reader as an increase in absorbance at 475 nm.

  At this time, the present inventors added the above sample dissolved in water or DMSO. However, the final concentration of DMSO was 0.1%.

As a method for calculating the inhibitory activity, the present inventors used the following formula to calculate the inhibition rate from the absorbance increase rate (control) when only the solvent was added and the absorbance increase rate (sample) when the above sample was added. The activity was calculated, and the 50% inhibitory concentration (IC ₅₀ ) was determined from the sample concentration and the inhibitory activity.

  Inhibitory activity (%) = 100− (sample / control) × 100

b) Test results Table 1 shows the results of the tyrosinase inhibition test.

c) Discussion As can be seen from the results of the tyrosinase inhibition test shown in Table 1, the buckwheat extract obtained in Production Example 2 showed a stronger tyrosinase inhibitory activity than arbutin used as a control. Arbutin is a natural (β-glucoside) glycoside that is also contained in plants such as cowberry and walrus. It has an excellent whitening effect by inhibiting tyrosinase, an enzyme important for melanin synthesis. It is supposed to be shown. Therefore, it is clear that the buckwheat extract obtained in Production Example 2 has a very strong tyrosinase inhibitory activity.

<Test Example 2> (Elastase inhibition test)
a) Test Method The inventors put 100 μL of the substrate N-Suc- (Ala) 3-pNA in a 96-well microplate, and samples such as buckwheat extract (50% ethanol extract) obtained in Production Example 2. Appropriate amounts of 10 μl of the solution and water (amount that gives a total volume of 200 μL during the reaction) were added, and the mixture was incubated at 37 ° C. for about 5 minutes. Next, the present inventors added 20 μL of elastase to start the reaction, measured the increase in absorbance at 410 nm at 37 ° C. with a microplate reader over time, and measured the reaction rate (Abs) for 5 to 10 minutes after the start of the reaction. / Hr).

  At that time, the present inventors added the above sample dissolved in water or DMSO. However, the final concentration of DMSO was 0.5%.

As a method for calculating the inhibitory activity, the present inventors used the following formula to calculate the inhibition rate from the absorbance increase rate (control) when only the solvent was added and the absorbance increase rate (sample) when the above sample was added. The activity was calculated, and the 50% inhibitory concentration (IC ₅₀ ) was determined from the sample concentration and the inhibitory activity.

  Inhibitory activity (%) = 100− (sample / control) × 100

b) Test results Table 2 shows the results of the elastase inhibition test.

c) Discussion As can be seen from the results of the elastase inhibition test shown in Table 2, the buckwheat extract obtained in Production Example 2 showed a stronger elastase inhibitory activity than the Clara extract used as a control. In addition, Clara (scientific name: Sophora flavescens Aiton) is a leguminous plant, and the extract has an elastase inhibitory action and is supposed to prevent skin aging (refer patent document 16). Therefore, it is clear that the buckwheat extract obtained in Production Example 2 has a very strong elastase inhibitory activity.

<Test Example 3>
The present inventors correspond to the peaks (1) to (5) in the HPLC analysis of FIG. 1 based on the extracted extract obtained in Production Example 3 so as to have the same content and the same component ratio as the extracted extract. A composition (mixed composition) in which the components orientin, isoorientin, vitexin, isovitexin and rutin were mixed was prepared. Then, the extract and the mixed composition of Production Example 3 were each subjected to HPLC analysis under the same conditions as the HPLC analysis of FIG. FIG. 4 shows an HPLC chromatogram of the extract of Production Example 3. FIG. 5 shows a chromatogram of the mixed composition.

  In addition, Table 3 shows numerical data of the component ratios and contents of the extract and the mixed composition of Production Example 3.

Next, according to Test Example 1, the present inventors conducted tyrosinase inhibition tests of the extract of Production Example 3 and the mixed composition, and compared tyrosinase inhibitory activities. As a result, the present inventors found that the IC ₅₀ value of the extract of Production Example 3 was 81 μg / ml, whereas the tyrosinase inhibitory activity of the mixed composition was very weak, about 15% even at 1000 μg / ml. It was found to be an inhibitory activity. In addition, as FIG. 6, the graph of the tyrosinase inhibitory activity test result of the extract of manufacture example 3 and a mixed composition is shown.

Next, the inventors conducted elastase inhibition tests of the extract and the mixed composition of Production Example 3 in accordance with Test Example 2, and compared the elastase inhibitory activity. As a result, the present inventors found that the extract ₅₀ of the production example 3 had an IC ₅₀ value of 323 μg / ml, whereas the elastase inhibitory activity of the mixed composition was very weak, about 3% even at 1000 μg / ml. It was found to be inhibitory activity. In addition, as FIG. 7, the graph of the elastase inhibitory activity test result of the extract of manufacture example 3 and a mixed composition is shown.

<Discussion>
As described above, the present inventors isolated five compounds from the fraction showing the tyrosinase inhibitory activity of buckwheat extract, and determined their structures by instrumental analysis such as HPLC. These compounds are all known components already isolated from buckwheat or buckwheat and have been reported to have antioxidant activity, but at least orientin and isoorientin show tyrosinase inhibitory activity. There are no reports. The present inventors have also confirmed that the buckwheat extract has elastase inhibitory activity.

  Furthermore, the present inventors have found that buckwheat extract is remarkably superior in both tyrosinase inhibitory activity and elastase inhibitory activity as compared with a simple mixed composition of five compounds contained in buckwheat extract. I found out.

  In addition, when compared to plant extracts used in other herbal medicines or herbs, the buckwheat extract has both tyrosinase inhibition and elastase inhibition compared to tyrosinase inhibitory activity and elastase inhibitory activity. Shows activity. There is no report that buckwheat has been used as a cosmetic so far.

  From the above, it is considered that buckwheat extract is useful as a new cosmetic material having an anti-aging effect excellent in both whitening and skin beautifying effects.

  In the above, this invention was demonstrated based on the Example. It is to be understood by those skilled in the art that this embodiment is merely an example, and that various modifications are possible and that such modifications are within the scope of the present invention.

  In the above examples, the extract of buckwheat extract is further filtered under reduced pressure to obtain an extract, and concentrated and freeze-dried to obtain a dry powder extract (in this specification, extract means an extract, not only a liquid However, it is not intended to be limited to a dry powder extract but may be used in a liquid form. . For example, it is possible to produce a moisturizing cream exhibiting a whitening action and a skin beautifying action (anti-aging action) by mixing the extract of buckwheat into a moisturizing cream which is a kind of cosmetics in a liquid state.

It is a HPLC chromatogram of the extract extracted from buckwheat. It is a graph which shows the result of having examined the extraction method and extraction efficiency from buckwheat. It is a figure explaining the structural formula and tyrosinase inhibitory activity of peaks 1-5. It is a HPLC chromatogram of the extract extracted from buckwheat. It is a chromatogram of a mixed composition. It is a graph which shows the result of the tyrosinase inhibitory activity test of the extract and mixed composition from buckwheat. It is a graph which shows the result of the elastase inhibitory activity test of the extract and mixed composition from buckwheat.

Claims (6)

A whitening agent comprising an extract of buckwheat containing orientin and isoorientin extracted with a water-containing organic solvent. The whitening agent according to claim 1,
The water-containing organic solvent is a whitening agent containing one or more organic solvents selected from the group consisting of ethanol, isopropanol, butylene glycol, and acetone. In the whitening agent according to claim 1 or 2,
The water-containing organic solvent is a whitening agent having an organic solvent content of 10% (v / v) or more and 90% (v / v) or less. In the whitening agent in any one of Claims 1 thru | or 3,
The extract is a whitening agent derived from an extract obtained by depositing buckwheat with water-containing ethanol under a temperature condition of 4 ° C. to 30 ° C. for a period of 1 day to 4 days. In the whitening agent in any one of Claims 1 thru | or 3,
The extract is a whitening agent derived from an extract obtained by heating buckwheat with water-containing ethanol under a temperature condition of 40 ° C. or more and a boiling temperature or less for a period of 0.5 hours to 2 hours. A tyrosinase inhibitor comprising a buckwheat extract containing orientin and isoorientin extracted by a water-containing organic solvent.