JP5531197B2 - Tyrosinase activity inhibitor and whitening cosmetic containing the same - Google Patents

Description

  The present invention relates to a tyrosinase activity inhibitor, and more particularly to a tyrosinase activity inhibitor that effectively inhibits tyrosinase related to melanin production and a whitening cosmetic using the same.

  Currently, many whitening cosmetics are on the market for the purpose of preventing sunburn, spots, freckles and the like on the skin. These whitening cosmetics contain components that prevent the production of melanin, which is a pigment substance, and many of them are tyrosinase activity inhibitors that inhibit tyrosinase, which is the rate-limiting enzyme for melanin production.

  As typical tyrosinase activity inhibitors, kojic acid, arbutin and the like are known. Of these, kojic acid is produced in the fermentation product of Aspergillus oryzae, and its tyrosinase inhibitory activity is high, but there are reports of carcinogenicity by oral ingestion and side effects such as leukocyte reduction, etc. It is hard to say that is sufficiently secured. Arbutin is a compound that can be obtained from an extract of walnut and bilberry, and its safety has been almost confirmed. However, there is a problem that its tyrosinase inhibitory activity is weaker than that of kojic acid.

Furthermore, although reports on obtaining tyrosinase activity-inhibiting components obtained from plants other than the above have been made (Patent Documents 1 to 6 and the like), there is still a strong demand for providing new tyrosinase activity-inhibiting components. there were.
JP-A-5-294819 JP-A-6-305978 JP 10-114670 A JP 2003-55189 A JP 2006-176420 A JP 2006-69954 A

  An object of the present invention is to provide a novel tyrosinase activity inhibitor having excellent activity and high safety.

  The inventors of the present invention focused on the plant resources of Okinawa Prefecture and conducted an extensive search for those having tyrosinase inhibitory activity. As a result, they found that some plants have excellent tyrosinase inhibitory activity and completed the present invention. .

  That is, the present invention is a tyrosinase activity inhibitor comprising as an active ingredient an extract of a plant selected from the group consisting of the genus Euphorbiaceae, the Rubiaceae, the Euphorbiaceae, and the Iranaceae.

  Moreover, this invention is the skin external preparation and whitening cosmetics containing the said tyrosinase tyrosinase activity inhibitor.

  An extract of a plant growing in Okinawa which is an active ingredient of the tyrosinase activity inhibitor of the present invention has excellent tyrosinase inhibitory activity and high safety.

  Therefore, the tyrosinase activity inhibitor of the present invention can be used as a whitening component in a skin external preparation, a whitening cosmetic, or the like.

  The active ingredient of the tyrosinase activity inhibitor according to the present invention is an extract obtained by subjecting a plant of the genus Euphorbiaceae, Rubiaceae, Euphorbiaceae, or Iranaceae (hereinafter referred to as “extracted plant”) to an extraction treatment. .

  Among the plants to be extracted, examples of the Acetaceae plant include canister (Lucuma nervosa A.DC.), sapodilla (Manilkara zapota (L.) P.van Royen), etc. In the case of a sapodilla, it is preferable to use the fruit.

  Examples of the myrtaceae plant include peach peach (Syzygium samarangense Merr. & Perry.), Lemon eucalyptus (Eucalyptus citriodora Hook), adek (Syzygium buxifolium) and the like, and it is preferable to use the leaves of them.

  Examples of the Rubiaceae plant include Bosoji (Psychotria rubra), Benidemari (Ixora coccinea L.), etc. Among them, it is preferable to use the leaves of the red clover, and the leaves and flowers of Benidomari.

  Examples of the Iroaceae plant include the Japanese white pine (Limonium wrightii (Hance) O.Kuntze f.albo-lutescens Hatsusima) and the turmeric islet (Limonium wrightii (Hance) O.Kuntze f.wrightii). It is preferable to use the above-ground part as a white pine pine and the rhizome as a turmeric pine.

  Examples of the genus Hygiaceae include red guinea pig (Bruguiera gymnorrhiza), Japanese leeches (Kandelia candel (L.) Druce), etc. As red leeches, the flowers and fruits are used, and as the leeches, the leaves are used. It is preferable.

  Examples of cynoaceae plants include Ryukyu Kokutan (Diospyros ferra var. Buxifolia Bakh), and examples of mulberry family plants include jackfruit (Artocarpus heterophyllus Lam.). It is preferable to use the leaves of each.

  Examples of the primrose plant include hamabos (Lysimachia mauritiana Lam.), Preferably using the whole plant, and the succulent plant (Lumnitzera racemosa Willd.) Is used and its leaves are used. It is preferable to do.

  Ginger (Alpinia speciosa (Wendl.) K. Schum) etc. are mentioned as a ginger family plant, It is preferable to utilize the flower, and Ryukyu Uchitorimochi (Balanophora kuroiwai Makino) etc. are mentioned as a plant family It is preferable to use the leaves.

  Examples of camellia plants include Adinandra ryukyuensis and the like, and it is preferable to use the leaves thereof, and examples of Euphorbiaceae plants include red chrysanthemum (Bischofia javanica Bl.) And the like are used. It is preferable.

  An example of the genus Acanthaceae is Mayopushiki (Sonneratia alba) and the like, and an example of the Amaryllidaceae plant is Craneum (Crinum asiaticum L. var japonicum aker), both of which preferably use the leaves.

  For example, for example, Koelreuteria elegance ACSm. Var.formosana FGMeyer, etc. It is preferable to use it.

  Examples of the liliaceae plant include the smeltax rose (Smilax sebeana Miq.) And the like, and it is preferable to use the ground part thereof.

  Examples of the extraction solvent for the plant to be extracted include lower alcohols such as water and ethanol, and a mixed solution of water and lower alcohol, particularly a mixture of water and ethanol is preferable.

  The extraction process of the plant to be extracted includes, for example, drying leaves, flowers, rhizomes, aerial parts or whole plants of the plant, then cutting or crushing them, and adding 5 to 20 times by weight of an extraction solvent thereto. In addition, it is carried out by immersing at room temperature to 100 ° C., preferably 30 to 50 ° C., for 12 to 150 hours, preferably 24 to 48 hours. In this extraction, it is preferable to shake or mechanically stir.

  Subsequently, this extract is clarified by solid-liquid separation means using industrial filter paper, diatomaceous earth, centrifugal separation, membrane filter or the like, whereby the plant extract used in the present invention can be obtained.

  The extract from the plant to be extracted can be further purified by using a known purification means, for example, appropriate various column chromatography, ultrafiltration, etc. as necessary to obtain a tyrosinase activity inhibitor. Moreover, it can also be set as the tyrosinase activity inhibitor pulverized by the well-known drying means, for example, spray drying, freeze drying, drum drying etc. as needed.

  The tyrosinase activity inhibitor of the present invention obtained as described above can be used by blending with a skin external preparation or a whitening cosmetic composition according to a conventional method.

  For example, in the case of whitening cosmetics, for example, about 0.001 to 5% by mass of the above tyrosinase activity inhibitor is blended with a known cosmetic base, and cosmetics, emulsions, creams, pastes are added according to conventional methods. And it is sufficient.

  EXAMPLES Next, although a reference example and an Example are given and this invention is demonstrated in more detail, this invention is not restrict | limited at all by these Examples.

Reference example 1
Obtaining extracts from extracted plants:
Extracted plants listed in Table 1 and Table 2 were obtained and subjected to the following extraction operation to obtain an extract of the extracted plant as a plant extract.

(Extraction operation)
Each plant to be extracted was cut into an appropriate size for each part and then dried at 60 ° C. for 12 hours, and pulverized using a centrifugal pulverizer (MRK-Retschm, ZM-100). What passed through a 0.5 mm screen was collected, and this was subjected to solvent extraction using a high-speed solvent extraction device (manufactured by Nippon Dionex, ASE-200). That is, each 2 g of sample by dry weight was loaded into an extraction cell together with 8 g of diatomaceous earth, and 50% ethanol solution was used to extract 50 ml in total at an extraction temperature of 80 ° C., an extraction time of 10 minutes, a pressure of 1500 psi, and two extractions. A plant extract was obtained.

Example 1
The plant extract obtained in Reference Example 1 was diluted 10 times with ion-exchanged water (hereinafter referred to as “sample solution”) and subjected to a tyrosinase inhibitory activity test. First, tyrosinase (derived from mushrooms, manufactured by Sigma) was adjusted to 40 U / ml with ion-exchanged water to obtain an enzyme solution. In addition, 3,4-dihydroxy-L-phenylalanine (hereinafter referred to as “DOPA”; manufactured by Sigma) was adjusted to 2.5 mM with ion-exchanged water to obtain a substrate solution. Furthermore, 1 / 15M phosphate buffer (pH 6.8) was used as a buffer solution.

  The enzyme reaction was performed in a 96-well microplate. To each well of the microplate, 100 μl of buffer solution, 40 μl of enzyme solution and 20 μl of sample solution were added, mixed well, and pre-incubated at 23 ° C. for 3 minutes. After pre-incubation, 50 μl of the substrate solution was added, and the absorbance at 490 nm (As0) was immediately measured. The enzyme reaction was performed at 23 ° C. for 10 minutes, and the absorbance (As10) after 10 minutes of the reaction was measured. Furthermore, the reaction was carried out in the same manner in a system in which 20 μl of a 5% ethanol solution was added instead of 20 μl of the sample solution and a system in which 40 μl of a buffer solution was added instead of 40 μl of the enzyme solution, and the absorbance immediately after the start of the reaction and after 10 minutes in each case (Ac0, Ac10 and Ab0, Ab10) were measured. Tyrosinase inhibitory activity was calculated by the following formula.

Tyrosinase inhibition rate (%) = 100 − [[(As10−Ab10) −
(As0−Ab0)] / (Ac10−Ac0) × 100]

Table 1 shows the tyrosinase inhibitory activity in each sample solution. In addition, the sample solution was dried under reduced pressure and re-dissolved in a 5% ethanol solution at a predetermined concentration. The concentration was decreased stepwise and the tyrosinase inhibitory activity test was conducted. The tyrosinase inhibitory activity was reduced to 50%. The concentration of tyrosinase in the sample solution (IC ₅₀ , mg / ml) was determined by interpolation. The results are shown in Table 2.

  As is clear from Tables 1 and 2, the plant extract of the present invention had excellent tyrosinase inhibitory activity.

Example 2
Whitening skin lotion (1):
A whitening lotion was prepared by the following composition and production method. This product had an excellent whitening effect.
(Composition) (mass%)
(1) Glycerin 5
(2) 1,3-butylene glycol 5
(3) POE (20) sorbitan monolaurate 1
(4) Ethanol 8
(5) Usujiro pine extract ^* 2
(6) Preservative appropriate amount (7) Perfume appropriate amount (8) Remaining amount of purified water
* Obtained in Reference Example 1

(Production method)
(B) (1) and (2) are sequentially dissolved in this order in (8) (b) (3), (5), (6), (7), and (9) are sequentially added in this order Dissolve (c) The aqueous solution obtained in (a) and the aqueous ethanol solution obtained in (b) using a vacuum emulsifier.
After mixing, the container was filled to obtain a translucent whitening lotion.

Example 3
Whitening skin lotion (2):
A whitening lotion was prepared by the following composition and production method. This product had an excellent whitening effect. (Composition) (mass%)
(1) 1,3-butylene glycol 5
(2) PEG 1500 8
(3) Sorbitol 7
(4) Hyaluronic acid 0.2
(5) Ethanol 7
(6) POE (5) oleyl alcohol ether 6
(7) Akagi extract ^** 1
(8) Olive oil 0.2
(9) Residual amount of purified water
** Obtained in Reference Example 1

(Production method)
(A) (1), (2), (3), (4) are sequentially dissolved in this order in (9). (B) (6), (7), (8) are sequentially dissolved in this order. Dissolve (c) The aqueous solution obtained in (a) and the ethanol solution obtained in (b) using a vacuum emulsifier.
After mixing, the container was filled to obtain a translucent whitening lotion (essence).

Example 4
Whitening skin lotion (3):
A whitening lotion was prepared by the following composition and production method. This product had an excellent whitening effect.
(Composition) (mass%)
(1) Dipropylene glycol 5
(2) PEG400 5
(3) Ethanol 7
(4) Ryukyu Tsuritori extract ⁺ 6
(5) Xanthan gum 0.2
(6) POE (6-20) sorbitan monostearic acid
Ester 1
(7) Oleyl alcohol 0.5
(8) Ascorbic acid 2
(9) Vitamin E acetate 0.2
(10) Residual amount of purified water
+ 50% ethanol extract from above ground

(Production method)
(B) (5), (1), (2), (8) are sequentially dissolved in this order in (10) (b) (7), (6), (4), (9) in (3) (C) The aqueous solution obtained in (a) and the ethanol solution obtained in (b) are dissolved using a vacuum emulsifier.
After mixing, the container was filled to obtain a milky whitening lotion (essence).

Example 5
Cosmetic oil:
A cosmetic oil was prepared by the following composition and production method. This product had an excellent whitening effect.
(Composition) (mass%)
(1) Olive oil 49
(2) Liquid paraffin 30
(3) Squalane 20
(4) Akabana hirugi extract ++ 0.9
(5) Vitamin E acetate 0.1
++ Extract of red berries of Reference Example 1

(Production method)
(B) Using (1) as a base, add (2), (3), (4), (5) to this and stir
(B) (i) was filtered and then filled into a container to obtain a cosmetic oil.

Example 6
Whitening cream:
A whitening cream was prepared by the following composition and production method. This product had an excellent whitening effect.
(Composition) (mass%)
(1) Glycerin 15
(2) 1,3 butylene glycol 4
(3) Potassium hydroxide 0.2
(4) Stearic acid 3
(5) Liquid paraffin 3
(6) Vaseline 7
(7) Stearic acid monoglyceride 3
(8) Lemon Eucalyptus Extract ^# 0.5
(9) Residual amount of purified water
#Things from Reference Example 1

(Production method)
(A) (1), (2), (3) are sequentially dissolved in this order in (9) (B) (4), (5), (6), (7), (8) After heating and melting, it was mixed using a vacuum emulsifier and further filled into a container to obtain a whitening cream.

  The extract from the plant to be extracted of the present invention has excellent tyrosinase inhibitory activity.

  Therefore, the tyrosinase activity inhibitor containing this extract as an active ingredient can be advantageously used as an active ingredient of a skin external preparation or a whitening cosmetic.

Claims (4)

An inhibitor of tyrosinase activity comprising an extract of Ryukyutsuritomochi as an active ingredient. Extract luchuensis Tutsi Birdlime is water and tyrosinase activity inhibitor according to claim 1, wherein the extract by a mixture of a lower alcohol. A skin external preparation containing the tyrosinase activity inhibitor according to claim 1 or 2 . A whitening cosmetic comprising the tyrosinase activity inhibitor according to claim 1 or 2 .