JP2012106971A - Melanin formation inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a melanin formation inhibitor capable of reducing the formation of the melanin in a skin cell.SOLUTION: The melanin formation inhibitor can eliminate (reduce) NO because of containing an extract of pine bark. The inhibitor containing the extract of the pine bark can reduce the formation of the melanin compared to that free from the extract of the bark.

Description

  The present invention relates to a melanin production inhibitor.

  In recent years, development of products having a function of reducing (erasing) active oxygen has progressed as awareness of beauty such as anti-staining and freckles has increased. This is due to the occurrence of spots and freckles due to the following action. That is, when the skin is exposed to ultraviolet rays, active oxygen is generated in the skin epidermis cells and dermis cells (fibroblasts, vascular endothelial cells, etc.). Then, the cells are damaged by the active oxygen, and various information transmission substances are generated and released. This various information transmission substances are caused by the production of melanin in skin cells and the generation of spots and freckles. In other words, if the active oxygen can be reduced (erased), the amount of melanin produced can be suppressed, and as a result, the generation of spots and freckles can be suppressed, and thus the products as described above have been developed.

JP 2006-282598 A

  In such a background, in addition to the technique described in Patent Document 1, development of a technique capable of suppressing the amount of melanin produced in skin cells has been desired.

  This invention responds to the above-mentioned request, and it aims at providing the melanin production amount inhibitor which can suppress the production amount of melanin in a skin cell.

  In order to achieve the above-mentioned object, the melanin production amount inhibitor according to claim 1 suppresses the production amount of melanin by containing a pine bark extract.

  The melanin production inhibitor according to claim 2 is the melanin production inhibitor according to claim 1, wherein the pine bark extract inhibits the activation of nitric oxide, thereby suppressing the production of melanin. Is.

  The melanin production inhibitor according to claim 3 is the melanin production inhibitor according to claim 1 or 2, wherein the pine bark extract is composed of an OPC content of 40% by mass or more.

  According to the present invention, there is an effect that the amount of melanin produced in the skin cells can be suppressed as compared with those not containing a pine bark extract. This effect will be described. One means for suppressing the amount of melanin produced is a means for reducing the action of α-melanocyte stimulating hormone (hereinafter referred to as “α-MSH”). In order to reduce the action of α-MSH, it is only necessary to reduce (eliminate) nitric oxide (hereinafter referred to as “NO”) having a function of enhancing the action. Here, the inventors have found that NO can be reduced (eliminated) by using a pine bark extract. Thereby, this invention containing a pine bark extract can suppress the production amount of melanin compared with what does not contain a pine bark extract.

It is the figure which showed the NO erasure | elimination activity rate in each raw material.

  Hereinafter, embodiments of the present invention will be described. The present invention is not construed as being limited by the description of the embodiments described later, and various modifications can be made within the scope of the claims.

  The melanin production amount inhibitor of the present invention is characterized by containing a pine bark extract to suppress the production amount of melanin. Moreover, the pine bark extract contained in the melanin production amount inhibitor is characterized by suppressing the production amount of melanin by eliminating nitric oxide. Furthermore, the pine bark extract contained in the melanin production rate inhibitor is characterized by comprising an OPC content of 40% by mass or more. In the present specification, a polymer having a degree of polymerization of 2 to 4 in proanthocyanidins is referred to as OPC (oligomeric proanthocyanidin).

  The pine bark extract used in the present invention contains proanthocyanidins as one of the main active ingredients. Proanthocyanidins refer to a group of compounds composed of a condensation polymer having a degree of polymerization of 2 or more having flavan-3-ol and / or flavan-3,4-diol as a constituent unit. Proanthocyanidins are powerful antioxidants produced by plants and are known to be found in plant leaves, bark, fruit peels and seeds. In addition, the pine bark extract used for this invention contains proanthocyanidins in the ratio of 30 mass%-60 mass%.

  The raw material of the pine bark extract used in the present invention is particularly limited, such as French coastal pine (Pinus Martina), larch, Japanese black pine, Japanese red pine, Japanese pine, Japanese pine, Korean pine, Japanese pine, Ryukyu pine, Japanese pine, Japanese pine, and Japanese white pine. Although not done, French coastal pine is preferred. The obtained extract may be concentrated or dried as necessary to form a liquid, a paste, or a powder. In addition, a pine bark extract manufactured by Toyo Shinyaku Co., Ltd. can be used particularly preferably.

  The pine bark extract used in the present invention contains a condensed polymer having a polymerization degree of 2 or more as proanthocyanidins. In particular, proanthocyanidins containing a large amount of a condensation polymer having a low degree of polymerization are preferred. The condensation polymer having a low degree of polymerization may be a condensation polymer having a degree of polymerization of 2 to 30 (2 to 30 mer), preferably a condensation polymer having a degree of polymerization of 2 to 10 (2 to 10 mer), A condensation polymer having a degree of polymerization of 2 to 4 (2 to 4 mer) is more preferable. The pine bark extract used in the present invention is an extract containing OPC in a proportion of 20% by mass or more, preferably in a proportion of 30% by mass or more, and more preferably in a proportion of 40% by mass or more. is there.

  The pine bark extract used in the present invention may further contain catechins. Catechin is a general term for polyhydroxyflavan-3-ol. As catechins, (+)-catechin (referred to as catechin in a narrow sense), (−)-epicatechin, (+)-gallocatechin, (−)-epigallocatechin, epigallocatechin gallate, epicatechin gallate, and afzelechin Etc. are known. In addition to (+)-catechin, gallocatechin, afzelekin, and (+)-catechin or 3-galloyl derivatives of gallocatechin have been isolated from pine bark extract.

  The pine bark extract is obtained by extracting pine bark with water or an organic solvent. When water is used, warm water or hot water is preferably used. Organic solvents used for extraction include foods or drugs such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, butane, acetone, hexane, cyclohexane, propylene glycol, hydrous ethanol, hydrous propylene glycol, etc. An organic solvent acceptable for the production of is preferably used. These water and organic solvents may be used alone or in combination of two or more solvents. In particular, water, ethanol, hydrous ethanol and the like are preferably used.

  The method for extracting the pine bark extract from the pine bark is not particularly limited, and for example, an extraction method such as a warm extraction method or a supercritical fluid extraction method can be used.

  In the method for extracting pine bark extract, a plurality of extraction methods may be combined. By combining a plurality of extraction methods, it becomes possible to obtain pine bark extracts having various compositions.

  The pine bark extract obtained by the above extraction can be eluted with an adsorbent or the like. The elution method is not particularly limited, but it is possible to obtain pine bark extracts having various compositions by combining a plurality of elution methods.

  The amount of pine bark extract used in the present invention is not particularly limited, but is preferably 5 mg or more, more preferably 20 mg or more per day. In addition, you may take the above-mentioned intake divided into several times a day.

  Regarding the mechanism by which the melanin production inhibitor of the present invention suppresses the production of melanin, “By removing (decreasing) NO in the pine bark extract, the action of α-MSH is reduced and the increase in tyrosinase is reduced. It is a mechanism that “suppresses and suppresses the amount of melanin produced”.

  In addition, the compounding amount of the melanin production amount inhibitor of the present invention is the type of product to be blended, age, sex, body weight, condition, administration time, dosage form, method of administration or ingestion, timing, etc. It can be set appropriately according to

  The melanin production inhibitor for oral administration or ingestion is in the form of capsules such as hard capsules and soft capsules, tablets and pills, or powders, granules, and candy according to the tastes of consumers. It can be formed into a shape. It can also be prepared in liquid dosage forms or forms such as solutions, suspensions, or emulsions.

  The melanin production inhibitor for oral administration or ingestion itself can be used as a pharmaceutical, a quasi-drug, a food for specified health use, a dietary supplement, other food or drink, or can be used in combination with these.

  The melanin production inhibitor for oral administration or ingestion, or a combination product thereof, may be eaten as it is depending on the dosage form, shape or preference, or dissolved in water, hot water, milk, soy milk, tea, juice, etc. You can drink it.

  Examples of the present invention will be described below. It should be noted that the present invention is not construed as being limited to the embodiments described later, and various modifications can be made within the scope of the claims.

NO can be eliminated by pine bark extract (it was demonstrated by the following test that it can be reduced. First, the four solutions shown in Table 1 (sample test solution, sample blank solution, control test solution and control blank) Solution) was produced for each material of pine bark extract, grape seed extract, dried pomegranate fruit extract, and “NO ₂ ⁻ + NO ₃ ⁻ ” measured values of 4 solutions in each of these materials were measured using the Griss method. Thereafter, the NO erasing activity rate of each material was calculated using Equation 1 described later.

  The sample test solution and the sample blank solution are solutions containing one material (sample) among the above-mentioned three materials, and the control test solution and the control blank solution are solutions not containing the material. .

(Table 1)

  Details of each material used for the measurement in the Griss method will be described. For each material, pine bark extract powder, grape seed extract powder (containing 81% or more proanthocyanidins), and dried pomegranate fruit extract powder were used. For these three materials, sample test solutions and sample blank solutions shown in Table 1 were produced by the following procedure. First, 10 mg of raw material was dissolved in 1 mL of 10% dimethyl sulfoxide. Next, the solution was diluted 10 times to produce a solution in which 1 mg of the material (sample) was dissolved in 1 mL of 2% dimethyl sulfoxide, and 60 μL of the solution was used as a sample test solution and a sample blank solution ( (See Table 1).

  Hereinafter, the sample test solution and sample blank solution in which the powder of pine bark extract is dissolved are referred to as “pine sample”, and the sample test solution and sample blank solution in which the grape seed extract powder is dissolved are referred to as “grape sample”. The sample test solution and the sample blank solution in which the dried pomegranate fruit extract powder is dissolved are referred to as “pomegranate sample”. In addition, 60 μL of a sample using only 2% dimethyl sulfoxide without dissolving the above-mentioned three raw material powders was produced as a control test solution and a control blank solution (see Table 1).

Next, measurement of “NO ₂ ⁻ + NO ₃ ⁻ ” in four solutions for each material using the Griss method will be described. For this measurement, “NO2 / NO3 Assay Kit-C II (Colorimetric)” manufactured by Dojindo Laboratories Co., Ltd. was used. Hereinafter, this is referred to as a “test kit”. In this measurement, five samples of “buffer solution, coenzyme, reductase, reagent A and reagent B” included in the test kit were used.

In the “NO ₂ ⁻ + NO ₃ ⁻ ” measurement in the four solutions described above, a case where a pine sample is used will be described as an example. First, “NO ₂ ⁻ + NO ₃ ⁻ ” measurement in the sample test solution will be described. “NO ₂ ⁻ + NO ₃ ⁻ ” measurement in the sample test solution was performed according to the procedure of the “sample test solution” part shown in Table 2.

(Table 2)

Specifically, 20 μL of 50 mM concentration of SNP (Sodium Nitroplusside) is added to 60 μL of a sample test solution in a pine sample, and the mixture is well mixed and left at room temperature for 30 minutes. Next, 10 μL of coenzyme and 10 μL of reductase are added to 80 μL of this solution and mixed well, and left at room temperature for 120 minutes. Then, 50 μL of reagent A is added to this solution and mixed well, and left at room temperature for 5 minutes. Thereafter, 50 μL of reagent B was added to the solution and mixed well, and left at room temperature for 10 minutes. Then, this solution was centrifuged at 10,000 rpm for 1 minute using a centrifuge, and the supernatant was added to a 96-well plate at 150 μL / well. Then, by measuring the absorbance at 540 nm using a microplate reader, “NO ₂ ⁻ + NO ₃ ⁻ ” of the sample test solution in the pine sample was measured.

Moreover, the “NO ₂ ⁻ + NO ₃ ⁻ ” measurement in the sample blank solution was performed according to the procedure of the “sample blank solution” part shown in Table 2. Specifically, 20 μL of a buffer solution is added to 60 μL of a sample blank solution in a pine sample and mixed well, and left at room temperature for 30 minutes. Next, 10 μL of coenzyme and 10 μL of reductase are added to 80 μL of this solution and mixed well, and left at room temperature for 120 minutes. Then, 50 μL of a buffer solution is added to this solution and mixed well, and left at room temperature for 5 minutes. Thereafter, 50 μL of a buffer solution was added to the solution, mixed well, and left at room temperature for 10 minutes. Then, this solution was centrifuged at 10,000 rpm for 1 minute using a centrifuge, and the supernatant was added to a 96-well plate at 150 μL / well. Then, by measuring the absorbance at 540 nm using a microplate reader, the “NO ₂ ⁻ + NO ₃ ⁻ ” of the sample blank solution in the pine sample was measured.

In the measurement of “NO ₂ ⁻ + NO ₃ ⁻ ” in the control test solution, the sample test solution shown in Table 2 was used except that 60 μL of the sample using only 2% dimethyl sulfoxide was used instead of 60 μL of the pine sample. Since it is the same procedure as the “NO ₂ ⁻ + NO ₃ ⁻ ” measurement, a description thereof will be omitted. Similarly, in the measurement of “NO ₂ ⁻ + NO ₃ ⁻ ” in the control blank solution, the sample blank solution shown in Table 2 was used except that 60 μL of the sample using only 2% dimethyl sulfoxide was used instead of 60 μL of the pine sample. Since this is the same procedure as the “NO ₂ ⁻ + NO ₃ ⁻ ” measurement in FIG.
Substitute each measured value of “NO ₂ ⁻ + NO ₃ ⁻ ” of the sample test solution, the sample blank solution, the control test solution, and the control blank solution in the pine sample, measured in the above-described procedure, into Equation 1 below. The NO elimination activity rate in the pine bark extract was calculated.

(Formula 1)
NO scavenging activity rate (%) = [1 - {(the sample test solution _"NO ² - + NO ₃ ^-" measurement - the sample blank solution _"NO ² - + NO ₃ ^-" measurements) of / (control test solutions, " NO ₂ ^- + NO ₃ ^- "measurement -" _NO ² control blank solution - + NO ₃ ^- "measurements)}] × 100

Incidentally, in the grape sample, sample test solution _"NO ^{2 -} + _{NO 3} -", ^"- + _{NO 3} - NO ^2" of the measurements and the sample blank solution for measurement, instead of pine sample 60 [mu] L, using grape sample 60 [mu] L Since others are the same as the case of the pine sample, description is abbreviate | omitted. Further, the NO elimination activity rate in the grape seed extract is obtained by calculating the measured value of “NO ₂ ⁻ + NO ₃ ⁻ ” of each solution (sample test solution, sample blank solution, control test solution and control blank solution) in the grape sample. Calculated by substituting into 1.

Further, in the pomegranate sample, sample test solution _"NO ^{2 -} + _{NO 3} -", ^"- + _{NO 3} - NO ^2" of the measurements and the sample blank solution for measurement, instead of pine sample 60 [mu] L, using a pomegranate sample 60 [mu] L Since others are the same as the case of the pine sample, description is abbreviate | omitted. The NO scavenging activity rate in the dried pomegranate fruit extract is the measured value of “NO ₂ ⁻ + NO ₃ ⁻ ” of each solution (sample test solution, sample blank solution, control test solution and control blank solution) in the pomegranate sample. It was calculated by substituting into Equation 1.

  Each NO elimination activity rate in the pine bark extract, grape seed extract and pomegranate fruit dried extract calculated as described above is shown in FIG. As can be seen from FIG. 1, the NO scavenging activity rate of the pine bark extract is 90.6%, compared with 39.7% of the dried pomegranate fruit extract, which is sometimes used as a measure against stains and freckles. The erasing activity rate is more than double. Further, the NO scavenging activity rate of the pine bark extract shows almost the same result as the value of the grape seed extract containing 81% or more of proanthocyanidins well known to exhibit the antioxidant function. From these results, the pine bark extract can achieve a high NO-eliminating activity rate substantially the same as that of the grape seed extract without containing 81% or more of proanthocyanidins like the grape seed extract, and the dried pomegranate fruit. It can be seen that a NO elimination activity rate superior to that of the extract can be realized.

  As described above, since the pine bark extract can eliminate NO (because it can be reduced), the melanin production inhibitor of the present invention containing the pine bark extract uses the pine bark extract. The amount of melanin produced can be suppressed as compared with the case of not.

  Since the melanin production amount inhibitor of the present invention contains a pine bark extract, NO can be eliminated (can be reduced). Therefore, this invention containing a pine bark extract can suppress the production amount of melanin compared with what does not contain a pine bark extract. Therefore, the melanin production amount inhibitor of the present invention can be used in industrial fields that require suppression of the melanin production amount.

Claims (3)

  A melanin production amount inhibitor characterized by containing a pine bark extract to suppress the production amount of melanin.   The said pine bark extract suppresses the production amount of melanin by eliminating nitric oxide, The melanin production amount inhibitor of Claim 1 characterized by the above-mentioned. The said pine bark extract is comprised by 40 mass% or more of OPC content, The melanin production amount inhibitor of Claim 1 or 2 characterized by the above-mentioned.

Images