JP5283873B2 - Whitening composition - Google Patents

Description

  The present invention relates to a whitening composition and use thereof. More specifically, the present invention relates to a whitening composition having an effect excellent in suppressing melanin deposition on the skin using lemon myrtle or an extract thereof, and use thereof.

  Pigmentation such as skin darkening, buckwheat, and skin darkening that occurs after sunburn is caused by markedly increased melanin production due to activation of pigment cells (melanocytes) present in the skin. Especially for middle-aged and elderly people, this is one of the serious skin problems.

  Melanin is changed from tyrosine to dopa, from dopa to dopaquinone by the action of tyrosinase, and further formed through an intermediate such as 5,6-dihydroxyindole. Tyrosinase is an enzyme that is biosynthesized in melanocytes. When melanocytes form melanin, dendrites called dendrites are elongated to send melanin to surrounding basal cells (keratinocytes). Thereby, melanin is supposed to be stored in keratinocytes.

  Therefore, in order to prevent and improve skin pigmentation, it is conceivable to inhibit the melanin production process or to lightly bleach already produced melanin.

  Based on such an idea, various whitening components have been conventionally proposed. For example, as a compound that inhibits tyrosinase activity and suppresses melanin production, sulfur compounds typified by glutathione can be mentioned. In addition, ascorbic acids such as hydrogen peroxide, hydroquinone, vitamin C and the like have been used to lightly bleach the produced melanin. As a method for reducing melanin in keratinocytes, local administration of vitamin A such as vitamin A acid has been performed.

  However, since sulfur compounds give off a remarkable off-flavor, it is inappropriate to use them for the purpose of whitening the skin. Since hydrogen peroxide and hydroquinone have safety problems such as stability and skin irritation, their use is generally limited. Ascorbic acid typified by vitamin C, when formulated as a component of a composition containing a large amount of moisture, such as a moist skin external preparation, is easily oxidized and unstable, causing discoloration and odor. Had.

  Under such circumstances, many attempts have been made to use natural products such as plant extracts as whitening components in consideration of safety and the like. In particular, plant-derived extracts also have the advantage of good image quality when blended into cosmetics.

By the way, as a function other than the above-described inhibition of the melanin production process and light-color bleaching of the produced melanin, it is also considered to obtain a whitening effect by inhibiting the extension of melanocyte dendrites and inhibiting the transport of melanin to keratinocytes. ing. For example, a dendrite elongation inhibitor composed of an extract of a plant belonging to the genus Asteraceae has been proposed (Patent Document 1).
JP 2004-250354 A (published September 9, 2004)

  As described above, various whitening components have been developed so far. However, in the field of beauty and the like, these whitening components are not sufficient, and development of a new whitening composition is strongly demanded.

  An object of the present invention is to provide a new whitening composition and an external preparation for skin using the same.

  In order to solve the above-mentioned problems, the present inventors have intensively studied. As a result, Lemon Myrtle has found a new fact that it has tyrosinase inhibitory activity and also has an inhibitory effect on melanocyte dendrite elongation, and has completed the present invention. One plant that has tyrosinase inhibitory activity and dendrite elongation inhibitory activity has never been reported before, and is a completely new finding.Lemon Myrtle is a herb with a dietary experience and is highly safe. is there. The present invention has been completed based on such novel findings, and includes the following inventions.

  That is, in order to solve the said subject, the composition for whitening which concerns on this invention is characterized by including at least one among a lemon myrtle and its extract.

  Furthermore, in the whitening composition according to the present invention, the extract is more preferably extracted from lemon myrtle using water or a hydrophilic solvent.

  Moreover, the skin external preparation which concerns on this invention is characterized by including the composition for whitening which concerns on the said invention.

  Moreover, the composition for eating and drinking according to the present invention is characterized by including the whitening composition according to the present invention.

  The whitening composition according to the present invention includes at least one of the lemon myrtle and the extract thereof as described above. Lemon Myrtle is a plant that has not been previously reported to contain a whitening component, and has the effect of providing a completely new whitening composition according to the present invention.

  An embodiment of the present invention will be described as follows.

<1: Whitening composition according to the present invention>
The whitening composition according to the present invention may include at least one of lemon myrtle and an extract thereof. Lemon Myrtle and its extract are excellent in whitening effect because they are excellent in tyrosinase activity inhibitory activity and melanocyte dendrite elongation inhibitory activity. Furthermore, since lemon myrtle is an herb with a dietary experience, the whitening composition according to the present invention has high safety.

  Lemon Myrtle is a kind of bush herb native to Australia, and its scientific name is Backhouse citriodora. Lemon Myrtle is known to be usable as soap or deodorant because it contains more citral having antibacterial and relaxation effects than lemon. Moreover, since the aromatic component of lemon myrtle is an oil-soluble substance, it has been used as herbal oil. However, the present inventors have found a new finding that has never been reported so far, that lemon myrtle, especially its extract, has an excellent whitening effect.

  The method for obtaining the lemon myrtle is not particularly limited and may be cultivated by a conventionally known method. For example, a naturally dried product of the lemon myrtle can be easily obtained.

  In the present specification, the “whitening composition” means a composition used for improving the deposition of a pigment such as melanin on the skin of a mammal. For example, a composition for whitening the skin of a mammal such as a human who has darkened a pigment such as melanin due to ultraviolet irradiation or aging is also included in the term “whitening composition”. The whitening composition according to the present invention exhibits a skin whitening effect by various mechanisms. That is, the whitening composition according to the present invention exhibits a whitening effect by inhibiting the melanin production process by inhibiting tyrosinase activity and the like, and inhibiting melanin transport by inhibiting dendritic elongation of melanocytes. The mechanism of whitening by the composition for use is not limited to this. It is not clear what components contained in lemon myrtle inhibit tyrosinase activity or dendrite elongation of melanocytes. It is not clear whether or not the substance having tyrosinase inhibitory activity in lemon myrtle and the substance having dendrite elongation inhibitory activity of melanocytes are the same, but as shown in the examples below, lemon myrtle has at least tyrosinase inhibition. It became clear that the substance which has activity, and the substance which has the dendrite elongation inhibitory activity of a melanocyte were contained.

  The form of the lemon myrtle contained in the whitening composition according to the present invention is not limited as long as at least one of the lemon myrtle and its extract is included. For example, the lemon myrtle shredded product, crushed material, and pulverized Products and extracts, and these concentrates and dried products may be mixed as appropriate. Especially, it is preferable that the composition for whitening which concerns on this invention contains the extract of lemon myrtle.

  The extract of lemon myrtle is not particularly limited as long as it is extracted from lemon myrtle, but from lemon myrtle, using water or a hydrophilic solvent (hereinafter referred to as “hydrophilic solvent etc.”), It is preferable that it is extracted. Since the extract extracted using a hydrophilic solvent or the like has high tyrosinase activity inhibitory activity and melanocyte dendrite elongation inhibitory activity, it can provide a whitening composition with more excellent whitening effect. As described above, Lemon Myrtle, although its oil-soluble component has been mainly used, the present inventors tried to extract with a hydrophilic solvent, the resulting extract, They have found that they are excellent in tyrosinase activity inhibitory activity and melanocyte dendrite elongation inhibitory activity.

  Examples of the hydrophilic solvent include water, methanol, ethanol, and n-propanol. These may be used alone or in combination of two or more. Of these, water is preferred. The extract obtained using water has high tyrosinase activity inhibitory activity and melanocyte dendrite elongation inhibitory activity. In addition, as described later, when the whitening composition according to the present invention is included in the composition for eating and drinking, it is preferable from the viewpoint of safety if water is used as a hydrophilic solvent or the like.

  The method for extracting lemon myrtle is not limited, and any conventionally known method such as a continuous method or a batch method may be used. The extraction conditions are not particularly limited, and the extraction may be performed by performing an immersion or digestion for an arbitrary time. For example, lemon myrtle and / or a dried product thereof may be immersed in a solvent at room temperature for several minutes to 7 days, preferably for 1 hour to 3 days. Moreover, a lemon myrtle and / or its dried product may be heated with a solvent for 3 to 12 hours, preferably 1 to 7 hours, or may be heated to reflux. Thus, it is preferable to perform extraction from a lemon myrtle by heat extraction. According to the heat extraction, the tyrosinase activity inhibitory activity of the obtained extract and the dendrite elongation inhibitory activity of melanocytes are further improved. Moreover, according to the heat extraction, an extract having a tyrosinase activity inhibitory activity and a melanocyte dendrite elongation inhibitory activity can be obtained more quickly and easily. If extraction is performed under subcritical conditions, the extraction efficiency is further improved.

  After the extraction, a process such as removing the extraction residue by filtration or the like may be performed. Moreover, you may concentrate the obtained extract by vacuum filtration or ultrafiltration further. Alternatively, the solvent may be completely removed to dry or freeze-dry. Such a concentrate or a substance obtained by drying or freeze-drying is also within the category of “extract” in the present specification.

  In addition, you may give processes, such as drying, shredding, and grinding | pulverization, before the extraction process to the lemon myrtle used for extraction.

  Moreover, the composition for whitening which concerns on this invention may show a relaxation effect with the aromatic component contained in the extract of lemon myrtle. Moreover, lemon myrtle and / or its extract are excellent in tyrosinase activity inhibitory activity and melanocyte dendrite elongation inhibitory activity as described above. That is, the composition containing at least one of lemon myrtle and an extract thereof can also be a melanin deposition inhibiting composition for inhibiting the deposition of melanin on the skin.

  The content of lemon myrtle and the extract thereof contained in the whitening composition according to the present invention is not particularly limited, but is 0.0001% by weight or more and 20% when the total amount of the whitening composition is 100% by weight. It is preferable to set it as the range of weight% or less, More preferably, it is 0.001 to 5 weight%. If it is this range, the more excellent whitening effect can be acquired, and sufficient effect sufficient for the quantity of the lemon myrtle and its extract contained in the composition for whitening can be acquired.

  In addition, components other than lemon myrtle and its extract contained in the whitening composition according to the present invention are not particularly limited. For example, oily ingredients, surfactants, and ultraviolet rays commonly used in whitening cosmetics. Components such as an absorbent, a lower alcohol, an antiseptic, a bactericidal agent, a colorant, a powder, a fragrance, a water-soluble polymer, a buffer, and a conventionally known whitening component can be appropriately blended within a range not impairing the effects of the present invention.

<2: External preparation for skin according to the present invention>
The skin external preparation according to the present invention may include the whitening composition according to the present invention.

  Since the external preparation for skin according to the present invention has an excellent whitening effect and includes the highly safe whitening composition according to the present invention, it can prevent and improve the occurrence and progression of pigmentation in the skin. It is excellent in the effect of whitening the skin. For example, the external preparation for skin according to the present invention can take the form of an external preparation for skin whitening that is effective for the prevention and / or treatment of spots, freckles and the like.

  The content of the whitening composition in the external preparation for skin according to the present invention is not particularly limited, but the total amount of the external preparation for skin is 100% by weight, and the total amount of lemon myrtle and its extract is the dry solid content of lemon myrtle The content is preferably in the range of 0.0001 wt% or more and 20 wt% or less, and more preferably 0.001 wt% or more and 5 wt% or less. If it is in this range, it is possible to obtain an excellent effect of preventing and improving pigmentation in the skin, and sufficient effect corresponding to the amount of lemon myrtle and its extract contained in the whitening composition. Can be obtained.

  The dosage form of the external preparation for skin according to the present invention is not particularly limited, and examples thereof include cream, emulsion, oil, lotion, ointment, pack, aqueous gel, oil gel and the like. Of these, creams, emulsions, oils, and the like are preferable because they are excellent in transdermal absorption. Further, medicinal cosmetics such as lotions, emulsions, creams, packs and soaps as quasi drugs, and skin external preparations such as lotions, emulsions, creams and ointments as pharmaceuticals are also included in the category of skin external preparations according to the present invention. It is. The method for producing these external preparations for skin is not particularly limited, and according to the dosage form, it is produced in the same manner as a conventionally known external preparation for skin, and the whitening composition according to the present invention is blended. Also good. Moreover, it is good also as a bath composition by mix | blending the whitening composition which concerns on this invention with a conventionally well-known bath agent.

  Ingredients other than the whitening composition according to the present invention contained in the external skin preparation according to the present invention are not particularly limited, and may include components blended in conventionally known external skin preparations. For example, water, hydrocarbons, esters, fats and oils, higher alcohols, higher fatty acids, surfactants, polyhydric alcohols, lower alcohols, preservatives, ultraviolet absorbers, antioxidants, thickeners, A moisturizer, powder component, anti-inflammatory agent, pH adjuster, sequestering agent, sugar, fragrance, colorant, various medicinal components, whitening component other than lemon myrtle and its extract may be included. These may be appropriately blended depending on the application of the external preparation for skin according to the present invention. Specific examples of these components are listed below, but the present invention is not limited to these. Moreover, the skin external preparation which concerns on this invention may contain the component illustrated below independently, and may contain it in mixture of 2 or more types.

  Examples of the hydrocarbons include liquid paraffin, petrolatum, squalane, ceresin wax, paraffin wax and the like.

  Examples of the esters include isopropyl myristate (IPM), synthetic gallow, jojoba oil, carnauba wax and the like.

  The above fats and oils include beef tallow, whale oil, seal oil, menhaden oil, hariba liver oil, cod liver oil, cod, tuna, turtle tallow, horse hoof, sheep hoof, mink, otter, marmot oil, and other animal fats, olive oil , Camellia oil, macadamia nut oil, olive oil, lanolin, castor oil, grape seed oil, cacao oil, palm oil, tree wax, jojoba oil, safflower oil, coconut oil, hazelnut oil, grape seed oil, avocado oil, soybean oil, etc. Examples thereof include synthetic fats and oils such as vegetable oils and silicone oils.

  Examples of the higher alcohols include cetanol, stearyl alcohol, capryl alcohol, lauryl alcohol, myristyl alcohol, cetyl alcohol, cholesterol, phytosterol, hexyldecanol, octyldodecanol, and behenyl alcohol.

  Examples of the higher fatty acids include stearic acid, oleic acid, capric acid, myristic acid, palmitic acid, behenic acid, lanolin fatty acid, linoleic acid, linolenic acid, lauric acid, and isostearic acid.

  Examples of the surfactant include anionic surfactants such as sodium lauryl sulfate and alkylsulfosuccinates, cationic surfactants such as quaternary alkylamine salts, polyoxyethylene cetyl ether, polyethylene glycol monostearate, and glyceryl monostearate. , Polyoxyethylene sorbitan monolaurate, polyoxyethylene lauryl ether, polyoxyethylene polyoxypropylene glycol, fatty acid monoglyceride, polyoxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearyl ether, polyoxyethylene Oleyl ether, polyoxyethylene higher alcohol ether, polyoxyalkyl allyl ether, polyoxyethylene distyrene Phenyl ether, polyoxyethylene derivative, sorbitan monolaurate, sorbitan monooleate, sorbitan sesquioleate, sorbitan monolaurate, polyoxyethylene monolaurate, polyoxyethylene sorbitan laurate, polyoxyethylene sorbitan monostearate , Polyoxyethylene sorbitan monooleate, 1-polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan tetraoleate, polyethylene glycol monolaurate, polyethylene glycol monooleate, polyoxythylene Nonionic surfactants such as castor oil, polyoxyethylene lanolin, alkylbetaines, glycine types, alkylaminobetaines, immi Gelsolin type, L- arginine and amphoteric surfactants L- lysine-type or the like, monostearate polyoxyethylene sorbitan, it can be exemplified a nonionic surfactant such as polyoxyethylene stearic acid amide.

  Examples of the polyhydric alcohols include glycerin, propylene glycol, and 1,3-butylene glycol.

  Examples of the lower alcohols include ethanol and propanol.

  Examples of the preservative include parabens, chlorhexidine gluconate, camphor, methyl paraoxybenzoate, benzoate, salicylic acid, phenoxyethanol and the like.

  Examples of the ultraviolet absorber include paraaminobenzoic acid derivatives, benzophenone derivatives, anthranilic acid ultraviolet absorbers, salicylic acid ultraviolet absorbers, cinnamic acid ultraviolet absorbers, and sugar ultraviolet absorbers. In place of or together with the ultraviolet absorber, an ultraviolet dispersant such as zinc oxide may be included.

  Examples of the antioxidant include vitamin E and butylhydroxytoluene (BHT).

  Examples of the thickener include water-soluble polysaccharides such as gum arabic, carboxyvinyl polymer, xanthan gum, polyvinyl alcohol, silicic anhydride, xanthan gum, water-soluble celluloses such as hydroxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose, pullulan, polyacrylic Examples thereof include water-soluble polymers such as sodium acid.

  Examples of the humectant include polyhydric alcohols such as polyethylene glycol, sugars such as glucose, sorbitol, dextrin, trehalose, and lactose and derivatives thereof, sodium glutamate, keratin derivatives, collagen derivatives, amino acids such as trimethylglycine and derivatives thereof, Water-soluble polymers such as carboxyvinyl polymer, sodium chondroitin sulfate, sodium hyaluronate, sodium pyrrolidone carboxylate, sodium lactate, seaweed extract, yeast extract, various plant extracts with moisturizing action, octyldodecyl myristate, isopropyl palmitate, myristine Esters such as isopropyl acid, octyldodecyl oleate, cholesteryl oleate, sodium polyacrylate, crystalline cellulose, various plant essential oils and these Mixtures thereof. Moreover, the above-mentioned vegetable oils and fats, fatty acids, and higher alcohols can also be used as a humectant.

  Examples of the powder component include titanium oxide, silica gel, talc, and acrylic acid-methacrylic acid copolymer.

  Examples of the anti-inflammatory agent include dipotassium glycyrrhizinate, tranexamic acid and derivatives thereof, and disodium 3-succinyloxyglycyrrhetinate.

  Examples of the pH adjuster include citrate, acetate and the like, sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid and the like as glucose as glucose. , Fructose, mannose, sucrose, trehalose, sulfated trehalose and the like.

  Examples of the fragrances include vanillin, orange flavor, lemon flavor, milk flavor geraniol, linalool and the like.

  Examples of the colorant include water-soluble tar dyes, water-insoluble tar dyes, gardenia dyes, safflower dyes, turmeric dyes, paprika dyes, anato dyes, cochineal dyes and other natural dyes, acidic and basic dyes. , Bengala and the like.

  Examples of the medicinal ingredients include medicinal ingredients according to the intended use of the external preparation for skin according to the present invention, such as hyaluronic acid, placenta extract, licorice extract, ginseng extract, vitamins, sterol glycosides and the like. Can do.

  Further, as whitening ingredients other than lemon myrtle and its extract, for example, vitamin C and its derivatives, arbutin and its derivatives, lucinol and its derivatives, hydroquinone and its derivatives, hydroquinone glycoside, tranexamic acid and its derivatives, salicylic acid and its Derivatives, kojic acid, ellagic acid, menthyl lactate, resorcin and derivatives thereof, menthol derivatives, lipoic acid, herbal medicine and extracts thereof, food materials and extracts thereof, and the like may be included.

  In addition to the components exemplified so far, the topical skin preparation according to the present invention includes caffeine, tannin, verapamil, grabridine, glycyrrhizic acid and derivatives thereof, or 4- (1,1-dimethylethyl) -4. '-Methoxy-dibenzoylmethane, potassium hydroxide and the like may be optionally blended. Further, the external preparation for skin according to the present invention is a wax (for example, microcrystalline wax, lanolin wax, candelilla wax, spermaceti, cocoa butter, silicon wax, oleate, myristate, linoleate, stearate, paraffin, beeswax, carnauba wax. , Ground wax, candelilla wax, microcrystalline wax, etc.).

  Moreover, the skin external preparation which concerns on this invention may contain the additive conventionally used for a conventionally well-known cosmetic composition other than the composition for whitening as needed. Examples of additives that can be used include collagen, hyaluronic acid, elastin, mucopolysaccharide vitamin A and its derivatives, vitamin B2, pantothenic acid and its derivatives, niacin, biotin and the like. Moreover, what is necessary is just to mix | blend the whitening composition which concerns on this invention in a conventionally well-known ointment, when making the dosage form of the skin external preparation which concerns on this invention ointment. Examples of conventionally known ointments include Macrogol 4000 and Macrogol 400.

  The use of the external preparation for skin according to the present invention is not particularly limited, and can be used for prevention and / or improvement of skin pigmentation. Examples thereof include pharmaceuticals, quasi drugs, and cosmetics. A more suitable application is cosmetics.

<3. Eating and drinking composition according to the present invention>
The composition for eating and drinking according to the present invention only needs to contain the whitening composition according to the present invention. According to the composition for eating and drinking according to the present invention, a whitening effect can be obtained from the body by ingesting lemon myrtle and / or its extract from the mouth.

  The content of the whitening composition in the composition for eating and drinking according to the present invention is not particularly limited, but the total amount of the composition for eating and drinking is 100% by weight, and the total amount of lemon myrtle and its extract is dried by lemon myrtle. The solid content is preferably 0.0001% by weight or more and 20% by weight or less, and more preferably 0.001% by weight or more and 5% by weight or less. If it is in this range, it is possible to obtain an excellent effect of preventing and improving pigmentation in the skin, and sufficient effect corresponding to the amount of lemon myrtle and its extract contained in the whitening composition. Can be obtained.

  Although it does not specifically limit as a provision form of the composition for eating and drinking which concerns on this invention, A foodstuff, a drink, or feed etc. can be illustrated, More specifically, processed grain products (processed flour products, processed starch products, premixes) Processed products, noodles, etc.), processed oils and fats (plastic oils, tempura oil, mayonnaise, etc.), processed soybean products (tofu, miso, etc.), processed meat products (ham, bacon, etc.), marine products, dairy products (raw materials) Milk, cream, yogurt, butter, cheese, etc.), processed vegetables and fruits (pastes, jams, etc.), confectionery (chocolate, biscuits, etc.), alcoholic beverages (sake, wine, whiskey, shochu, beer, etc.) Beverages (soft drinks such as tea and coffee, drinks, etc.), seasonings, semi-dried or concentrated foods, dried foods, frozen foods, solid foods, liquid foods (soups, etc.) can be exemplified. In addition, forms that can be taken orally, such as tablets in the form of tablets, granules, capsules, etc. are also within the category of the composition for eating and drinking according to the present invention.

  As a manufacturing method of the food-drinking composition which concerns on this invention, even if it manufactures by the method similar to a conventionally well-known food-drinking composition according to the provision form, it mix | blends the whitening composition which concerns on this invention. Good.

Ingredients other than the whitening composition according to the present invention contained in the composition for eating and drinking according to the present invention,
In addition, the whitening composition according to the present invention can be used for food and drink by blending lemon myrtle and / or an extract thereof with ingredients included in conventionally known food and drink compositions such as tablets such as supplements, tea, and food. It is good also as a composition. Such a component is not limited as long as it is a conventionally known beverage or food component, and sugars such as glucose, surfactants, thickeners, preservatives, fragrances, coloring agents, nutrients such as vitamins or citric acid. An agent or the like may be added.

  Examples will be shown below, and the embodiments of the present invention will be described in more detail. Of course, the present invention is not limited to the following examples, and it goes without saying that various aspects are possible in detail. Further, the present invention is not limited to the above-described embodiments, and various modifications can be made within the scope shown in the claims, and the present invention is also applied to the embodiments obtained by appropriately combining the disclosed technical means. It is included in the technical scope of the invention. Moreover, all the patent documents described in this specification are used as reference in this specification.

<Example 1: Production of Lemon Myrtle Extract>
Extraction was performed by adding 3 g of dried lemon myrtle leaves to 40 ml of water and heating at 70 ° C. for 1 hour. Next, after cooling to room temperature, insoluble matters were removed by filtration. Thereafter, the mixture was concentrated under reduced pressure and then freeze-dried to obtain an extract.

<Example 2: Confirmation of melanin production inhibitory effect>
About the melanin production suppression effect (it is also called blackening suppression) of the extract of lemon myrtle, it confirmed using the cell pellet of the mouse melanoma cell.

First, 2 × 10 ⁴ B16 melanoma cells were seeded in a 35 mm diameter petri dish containing 3 ml of Eagle's minimal nutrient medium containing 10% (v / v) fetal calf serum, and 5% (v / v) carbon dioxide gas. Using a carbon dioxide incubator adjusted to 37 ° C., the cells were cultured at 37 ° C. for about 24 hours.

  Next, pure water or a solution obtained by dissolving an extract of lemon myrtle in pure water so that the final concentration was 0.08% (w / v) and 0.15% (w / v) was added to the petri dish. . Next, the cells were further cultured for 5 days under the same conditions as before adding pure water or the solution.

  Next, after trypsin treatment, the cells were collected in a 1.5 ml tube (manufactured by Eppendorf) by centrifugation, and the degree of whitening was visually evaluated. As a result of the evaluation, it was determined that the large whitening was ++, the whitening was +, the whitening was slightly +, and the whitening was not. At the same time, changes in the cell mass volume were visually determined and used as an index of cytotoxicity. The results are shown in Table 1.

As shown in Table 1, the lemon myrtle extract was shown to strongly suppress blackening, and no change in cell mass volume was observed. From this, it became clear that the extract of Lemon Myrtle has low toxicity and high whitening effect.
<Example 3: Confirmation of melanin production inhibitory effect>
Next, the confirmation test of melanin production suppression was performed using human melanocytes (manufactured by Kurabo Industries).

First, normal human melanocytes (manufactured by Kurabo Industries) were amplified with Medium 254 medium (human melanocyte medium supplemented with HMGS, Kurabo Industries). Amplified human normal melanocytes were added to a 96-well plate so that the number thereof was 10 ⁴ per well, and cultured in a carbon dioxide incubator adjusted to 5% (v / v) carbon dioxide at 37 ° C. for about 24 hours. Next, pure water or a solution in which the extract was dissolved in pure water so as to have a final concentration of 0.08 and 0.15% (w / v) was added to the petri dish. Then, after further culturing for 5 days under the same conditions as before adding pure water or the solution, the cells were recovered.

  Next, 50 μl of a cell suspension solution (PBS buffer containing Triton X-100 0.1% (v / v)) was added to the collected cells, and the cells were sufficiently suspended. Let stand for hours. Thereafter, 50 μl of 10 mM L-DOPA (manufactured by Nacalai) was added and incubated at 37 ° C. for 1 hour.

  Thereafter, the absorbance at 495 nm was measured with a plate reader (Multiscan Plus MKII, manufactured by Nippon Flow Laboratory Co., Ltd.), and the amount of melanin produced was measured (the value when the lemon myrtle extract was not added was taken as 100, relative values) ) The results are shown in Table 2.

  As shown in Table 2, the extract of lemon myrtle was shown to suppress melanin production even at low concentrations.

<Example 4: Relationship between extraction conditions and tyrosinase inhibitory activity>
In this example, in addition to the extract obtained in Example 1, the extract obtained by setting the temperature condition of extraction to 100 ° C. As a solvent for extraction, instead of water, a 50% ethanol solution, 100% ethanol The tyrosinase inhibitory activity of the extract obtained by using a solution, 100% methanol solution or chloroform was confirmed.

  First, a lemon myrtle extract was obtained by the same method as that described in Example 1 except for the above temperature conditions and the type of solvent.

Next, 1 ml of the cell suspension used in Example 3 was suspended so as to contain 1 × 10 ¹⁰ human melanocytes. Each extract was added to a solution in which 10 mM L-DOPA as a substrate for tyrosinase was dissolved in 100 μl of the suspended liquid so as to be 0.15% (w / v), and then at 37 ° C. for 1 hour. Left to stand. As a control, the same operation was performed when no extract was added.

  Next, the absorbance of each solution was measured at 495 nm with a plate reader (manufactured by Nippon Flow Laboratories, product name: Multiscan Plus MKII), and the absorbance of each sample when the absorbance of the control was 100 was calculated. That is, the tyrosinase activity of each solution was calculated with the tyrosinase activity in the control as 100%, and the difference in tyrosinase activity between the control and each solution was evaluated as the inhibitory activity of tyrosinase. The results are shown in FIG. FIG. 1 is a graph showing the results of measuring tyrosinase inhibitory activity. In FIG. 1, the horizontal axis indicates the type of solvent used for extraction, and the vertical axis indicates the relative value of tyrosinase activity when the control is 100.

  As shown in FIG. 1, the extract extracted with water at 70 ° C. showed the highest tyrosinase inhibitory activity.

<Example 5: Relationship between extract concentration and tyrosinase inhibitory activity>
Extracts obtained by the same method as described in Example 1 were 0% (w / v), 0.08% (w / v), 0.15% (w / v), 0.30% ( w / v), 0.60% (w / v), 1.20% (w / v), except that it was added (respectively referred to as samples 1 to 6), the method described in Example 4 Tyrosinase inhibitory activity was measured by the same method. As a control, the same operation was performed when no extract was added.

  As for tyrosinase inhibitory activity, the tyrosinase activity in each control was measured as tyrosinase activity in the control as in Example 4, and the difference in tyrosinase activity between the control and each solution was evaluated as tyrosinase inhibitory activity. did. The results are shown in FIG. FIG. 2 is a diagram showing the results of measuring tyrosinase inhibitory activity. In FIG. 2, the horizontal axis indicates the sample number. This number corresponds to the numbers shown in Table 3. The vertical axis shows the relative value of tyrosinase activity when the control is 100.

  As shown in FIG. 2 and Table 3, when the concentration of the extract was 0.15% (w / v), the tyrosinase activity was as high as 10%, that is, the tyrosinase inhibitory activity was as high as 90%. In addition, even when the concentration of the extract was 0.30% (w / v), 0.60% (w / v), and 1.20% (w / v), extremely high tyrosinase inhibitory activity was exhibited. However, it was also affected by the color of the extract itself. If a measurement method that is not affected by the color of the extract itself is used, higher tyrosinase inhibitory activity may be exhibited.

<Example 6: Confirmation of dendrite elongation inhibitory activity>
In this example, the morphological change of human melanocytes, specifically, dendrite elongation inhibitory activity was confirmed.

First, normal human melanocytes (manufactured by Kurabo Industries) were amplified with Medium 254 medium (human melanocyte medium supplemented with HMGS, Kurabo Industries). Amplified normal human melanocytes were added to a 48-well plate at 3 × 10 ³ per well and cultured in a carbon dioxide incubator adjusted to 5% (v / v) carbon dioxide at 37 ° C. for about 24 hours. did.

  Next, add pure water or a solution of Lemon Myrtle extract dissolved in pure water to a final concentration of 0.08% (w / v) and 0.15% (w / v). did. Then, after further culturing for 2 days under the same conditions as before adding pure water or the solution, the appearance of dendrites was visually observed. Those having large elongation inhibitory activity were evaluated as ++, those having moderate elongation as +, and those having none as-. The results are shown in Table 4.

  In all of the lemon myrtle extract addition groups, dendrite elongation was suppressed.

<Example 7: Production of whitening cream>
First, the following materials A to E were prepared.
A: A mixture of 1.00 g of lemon myrtle extract obtained by the method of Example 1 and 5.00 g of purified water B: 0.05 g of disodium 3-succinyloxyglycyrrhetinate
C: Squalane 10.00 g, octyldodecyl myristate 8.00 g, microcrystalline wax 4.00 g, behenyl alcohol 3.00 g, lipophilic glyceryl monostearate 2.50 g and monostearate polyoxyethylene sorbitan (20E.O. ) 2.50 g of mixture D: 1, 3-butylene glycol 10.00 g, methyl paraoxybenzoate 0.10 g and purified water 54.00 g of mixture E: perfume 0.30 g.

  Next, B was added to D heated to 80 to 85 ° C., and C dissolved by heating at 80 to 85 ° C. was added with stirring with a homomixer to uniformly emulsify. This was slowly cooled to about 50 ° C. at room temperature, after which A was suspended and mixture E was added. Furthermore, whitening cream was obtained by cooling to room temperature, continuing stirring.

<Example 8: Production of lotion>
First, the following materials F to I were prepared.
F: Mixture G of 1.30 g of zinc oxide, 1.10 g of anhydrous silicic acid, 2.00 g of talc, 0.01 g of bengara and 0.05 g of polyoxyethylene stearamide (4EO) G: In the method of Example 1 Mixture H of 0.60 g of the obtained lemon myrtle extract, 5.00 g of ethanol, and 5.00 g of purified water: Mixture I of 3.00 g of concentrated glycerin, 0.10 g of camphor, 0.05 g of methyl paraoxybenzoate and 0.05 g of fragrance : 81.74 g of purified water.

  Next, about 60 g of I was added to F and dispersed uniformly with a homomixer to prepare a powder dispersion. To this was added G solution and H, and the remainder of I was added. Next, a carmine lotion was obtained by uniformly dispersing with a homomixer.

<Example 9: Production of whitening ointment>
First, the following mixture of J and K was prepared.
J: Mixture of 47.50 g of Macrogol 4000 and 47.50 g of Macrogol 400 K: A solution obtained by dissolving 0.50 g of Lemon Myrtle extract obtained by the method of Example 1 in 4.50 g of purified water.

  Next, J was heated to 65 ° C. on a water bath and dissolved, and mixed uniformly to prepare a macrogol ointment base. This was kneaded to produce a whitening ointment.

  The whitening composition according to the present invention can prevent and improve skin pigmentation and is highly safe, and thus can be suitably used in the cosmetic industry and the like.

In the Example of this invention, it is a figure which shows the result of having measured the tyrosinase inhibitory activity of the extract of lemon myrtle. In the Example of this invention, it is a figure which shows the result of having measured the tyrosinase inhibitory activity of the extract of lemon myrtle.

Claims (2)

  A whitening composition comprising at least one of lemon myrtle and an extract thereof.   The whitening composition according to claim 1, wherein the extract is extracted from lemon myrtle using water or a hydrophilic solvent.

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