JP7387951B2 - Skin external preparations - Google Patents

Description

TECHNICAL FIELD The present invention relates to an external preparation for skin that has excellent antioxidative effects.

In general, antioxidant action is the action of suppressing or stopping chain reactions by suppressing the production of active oxygen and free radicals, which are the causative agents of oxidative stress disorders, and by capturing them. These active oxygen species and free radicals are byproducts of respiration in living organisms, and are also produced by physical stress and mental stress. Living organisms are originally equipped with functions such as SOD (superoxide dismutase) that suppress the production of these substances, but as we age, this suppressing function declines, and oxidation of the living body progresses, leading to various diseases as an aging phenomenon. Connect. Since the environment of the skin is different from that of other organs, the effects of oxidation from the outside air, ultraviolet rays, and resident bacteria on the skin are significant. In particular, aging phenomena caused by ultraviolet rays are distinguished by the name "photoaging," and it has been found that various aging phenomena, from sunburn to wrinkles, age spots, and dullness, are related to active oxygen calculated from ultraviolet rays. There is. Therefore, it is important not only to ingest substances with antioxidant effects through meals, but also to apply them externally to the skin in order to prevent aging and diseases.

Therefore, active oxygen scavengers and antioxidants have been studied for the purpose of protecting against damage caused by active oxygen, and foods, cosmetics, quasi-drugs, and other products containing active oxygen scavengers and antioxidants such as SOD-like active substances have been developed. Pharmaceuticals, etc. are being developed. For example, Patent Document 1 discloses a cosmetic containing nicotinamide mononucleotide as a characteristic ingredient. However, nicotinamide mononucleotide has the drawback that a satisfactory moisturizing effect cannot be obtained when it is used alone (see Patent Document 2).

In addition, Non-Patent Document 1 discloses that tulip flowers are extracted from tulip flowers for the purpose of effective utilization of the tulip flowers, which are grown during the production process of tulip bulbs and many of them are processed by plowing into the field. The antioxidant activity evaluation of tulip extract as a cosmetic raw material is disclosed.

However, the combination of nicotinamide mononucleotide and tulip extract has not been studied in the past as an external skin preparation with antioxidant effects.

Japanese Patent Application Publication No. 2016-135750 Japanese Patent Application Publication No. 2018-145167

Ayaka Ota, “2020 Graduation Thesis, Use of Tulip Extract as a Cosmetic Raw Material,” Toyama Prefectural University, Faculty of Engineering, Department of Biotechnology, 2021

In view of the above-mentioned circumstances, the present inventors conducted extensive studies and found that an excellent antioxidant effect can be obtained when nicotinamide mononucleotide and tulip extract extracted from tulip flowers are used together.

An object of the present invention is to provide a novel skin preparation with excellent antioxidative effects.

The skin external preparation according to claim 1 of the present invention is characterized by containing a tulip extract extracted from the flower part of a tulip and nicotinamide mononucleotide.

The skin external preparation according to claim 2 of the present invention is characterized in that it contains the tulip extract and the nicotinamide mononucleotide at 0.0001% to 10% by weight, respectively.

The skin external preparation according to claim 3 of the present invention is characterized in that the variety of the tulip is Huang Komachi, and the extraction method of the tulip extract is heating extraction or normal temperature extraction.

The skin external preparation according to claim 4 of the present invention is characterized in that the extraction solvent for the tulip extract is water, 1,3-butylene glycol, or ethanol.

According to the skin external preparation according to claim 1 of the present invention, by containing tulip extract extracted from tulip flowers and nicotinamide mononucleotide, the SOD-like activity rate can be increased, and the antioxidant Excellent action.

According to the skin external preparation according to claim 2 of the present invention, the SOD-like activity rate can be increased by containing 0.0001% to 10% by weight of tulip extract and nicotinamide mononucleotide, respectively. , has excellent antioxidant effects.

According to the external preparation for skin according to claim 3 of the present invention, since the tulip variety is Huang Komachi, the SOD-like activity rate can be significantly increased and the antioxidative effect is excellent. Further, since the tulip extract is extracted by heating extraction or room temperature extraction, the tulip extract can be extracted by various methods without depending on temperature, resulting in excellent productivity.

According to the external preparation for skin according to claim 4 of the present invention, since the extraction solvent for tulip extract is water, 1,3-butylene glycol or ethanol, tulip extract can be extracted easily and productivity can be improved. Excellent in

Hereinafter, in order to explain the present invention in detail, a production example of a tulip extract used in the present invention, a formulation example of a skin external preparation of the present invention, and an experimental example are given as examples, but the present invention is not limited thereto. do not have. Moreover, % shown in Examples indicates weight % unless otherwise noted.

The tulip extract used in the present invention is extracted from the flower part of Tulipa (scientific name: Tulipa L.), which belongs to the family Liliaceae, genus Tulipa. Examples of tulip varieties include Kōkomachi, Tateyama no Haru, Strong Gold, Amethyst, and Strawberry Star. The extraction method for tulip extract is not particularly limited, and for example, it may be heated or extracted at room temperature.

Examples of solvents for extracting tulip extract include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohol (1,3-butylene glycol, etc.) , propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, etc.) , propyl ether, etc.). Polar solvents such as water, lower alcohols and liquid polyhydric alcohols are preferred, and water, ethanol and 1,3-butylene glycol are particularly preferred. These solvents may be used alone or in combination of two or more.

The above-mentioned tulip extract may be used as an extracted solution, or may be used after concentration, dilution and filtration treatment, decolorization with activated carbon, etc., deodorization treatment, etc., as necessary. Furthermore, the extracted solution may be subjected to treatments such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.

Nicotinamide mononucleotide (chemical formula: C ₁₁ H ₁₅ N ₂ O ₈ P) used in the present invention is a compound that is produced in the bodies of many organisms including humans. It is generally called NMN (Nicotinamide mononucleotide) and is known as an intermediate metabolite involved in the biosynthesis of the coenzyme NAD+. Nicotinamide mononucleotide has two types of optical isomers, α-form and β-form, and the β-form is used in the present invention. Nicotinamide mononucleotide can be obtained, for example, by synthesizing nicotinamide riboside from nicotinamide and ribose, and then phosphorylating the 5-hydroxyl group of the ribose moiety. Specifically, for example, first, nicotinamide and L-ribose tetraacetate are dissolved in anhydrous acetonitrile, and after adding an excess amount of trimethylsilyltrifluorosulfonic acid under a nitrogen stream, the mixture is stirred at room temperature, and methanol is added. The reaction solution, in which the reaction has been stopped, is applied to a column packed with activated carbon, washed with distilled water, and then eluted with methanol to recover the product. Next, in order to perform a phosphorylation reaction of the 5-position hydroxyl group of the L-ribose moiety of this product, the above product was dissolved in trimethoxyphosphoric acid, phosphorus oxychloride was added dropwise under ice cooling, and the mixture was dissolved under a nitrogen stream. A cold acetonitrile-ether solution is added to the above reaction solution, which has been stirred and neutralized by adding an aqueous sodium hydroxide solution to stop the reaction. Thereafter, the lower layer (aqueous phase) is passed through an anion exchange resin to recover the reactant, and further purified using a cation exchange resin, whereby highly pure nicotinamide mononucleotide can be recovered. Moreover, nicotinamide mononucleotide is commercially available, and these commercial products can be purchased and used.

The above-mentioned tulip extract may be used as is in the external preparation of the present invention, and ingredients used in cosmetics, quasi-drugs, pharmaceuticals, foods, etc. may be used as long as the effects of tulip extract and nicotinamide mononucleotide are not impaired. fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, humectants, powders, ultraviolet absorbers, thickeners Components such as pigments, antioxidants, whitening agents, chelating agents, excipients, and coating agents can also be blended.

The dosage forms of the present invention include, for example, lotions, creams, massage creams, milky lotions, gels, aerosols, packs, detergents, bath preparations, foundations, powders, lipsticks, ointments, poultices, pastes, and plasters. , essences, powders, pills, tablets, injections, suppositories, emulsions, capsules, granules, liquids (including tinctures, liquid extracts, spirits, suspensions, lemonades, etc.), etc. .

The amount of the tulip extract used in the present invention is preferably 0.0001% by weight or more, more preferably 0.0001 to 10% by weight, based on the total amount of the external preparation. Furthermore, 0.0001 to 0.10% by weight is most preferred. If it is less than 0.0001% by weight, it is difficult to expect sufficient effects. If the amount exceeds 10% by weight, it is difficult to notice any enhancement of the effect and it is uneconomical.

The amount of the nicotinamide nucleotide used in the present invention is preferably 0.0001% by weight or more, more preferably 0.0001 to 10% by weight, based on the total amount of the external preparation. Furthermore, 0.0001 to 0.10% by weight is most preferred. If it is less than 0.0001% by weight, it is difficult to expect sufficient effects. If the amount exceeds 10% by weight, it is difficult to notice any enhancement of the effect and it is uneconomical.

[Example 1]

[Production Example 1] Room temperature extraction 30g (fresh weight) of tulip flower part of Kōkomachi variety was ground with a hammer in liquid nitrogen, 2.4 times the weight of purified water was added, and 50% by weight of 1,3-butylene glycol was added. 72 mL of the aqueous solution was added, and after standing at room temperature for one week, it was filtered, and the filtrate was left standing at room temperature again for one week, and then further centrifuged and filtered to obtain a tulip extract.
[Production Example 2] Direct steam distillation 30 g (fresh weight) of tulip flower parts of the Kōkomachi variety were ground with a hammer in liquid nitrogen, 3 times the weight of purified water was added, and a mantle heater was heated to control the temperature around 105°C. Steam distillation was performed using Distillation was continued until half the amount of purified water added was obtained. The obtained distillation and distillation residue were each left for one week at room temperature, and then centrifuged and filtered to obtain a tulip extract.
[Production Example 3] Heating and refluxing 30 g (fresh weight) of tulip flower parts of the Kōkomachi variety were ground with a hammer in liquid nitrogen, 2.4 times the weight of purified water was added, and the mixture was heated in an autoclave at 105°C for 6 hours. Heated. Thereafter, it was filtered and the resulting filtrate was divided in half. A filtration fraction containing only the filtrate and a flower fraction containing the filtrate were prepared by adding the filtrate to half of the filtrate residue, and 40% (v/v) of 1,3-butylene glycol was added to each. After standing for one week at room temperature, each sample was refiltered to obtain a tulip extract.

Next, an experimental example will be given to explain the effects of the present invention in detail.

[Experimental Example 1] Evaluation of SOD (superoxide dismutase)-like activity Four types of lotions were prepared based on the four formulations shown in Table 1, and the superoxide anion scavenging ability was measured by calculating the SOD-like activity rate. Regarding the test concentration, the sample was serially diluted with ultrapure water to adjust a total of 7 concentrations, and the test was conducted. In addition, the superoxide anion scavenging ability was measured using SOD Assay Kit WST (Cat No. S311, Dojindo, Japan). The nicotinamide mononucleotide manufactured by Access One Co., Ltd. was used.
For details on the evaluation method, first, 20 μL of the specimen or control, 200 μL of WST working solution, and 20 μL of Enzyme working solution were added to a 96-well plate. In addition, Dilution buffer was used as a blank instead of Enzyme working solution. Three wells were used for one treatment group. The 96-well plate was then sealed with a plate seal, shaken at 270 rpm for 10 seconds, and incubated at 37°C for 20 minutes. Next, absorbance at 450 nm was measured using a microplate reader. Then, the superoxide anion residual rate and scavenging rate of the test product were calculated from the absorbance of the test product and the control using the following formula, and were determined as the SOD-like activity rate of the test product.

[Math 1] Superoxide anion residual rate (%)
=(S-SB)/(C-CB)×100

[Math 2] Superoxide anion elimination rate (%)
= {(C-CB)-(S-SB)}/(C-CB)×100
= SOD-like activity rate (%)

C: Absorbance of control CB: Blank absorbance of control S: Absorbance of test product SB: Blank absorbance of test product

The results of these experiments are shown in Tables 2 to 5, FIGS. 1 and 2. Table 2 shows the results of the SOD-like activity evaluation test for Comparative Formulation 1 (blank) in Table 1. Table 3 shows the results of the SOD-like activity evaluation test for Comparative Formulation 2 (containing 10% tulip extract) in Table 1. Table 4 shows the results of the SOD-like activity rate evaluation test for Comparative Formulation 3 (containing 10% nicotinamide mononucleotide (NMN)) in Table 1. Table 5 shows the results of the SOD-like activity evaluation test for Formulation 1 in Table 1 (containing 10% tulip extract and 10% nicotinamide mononucleotide (NMN)). In addition, Figure 1 is a diagram showing the SOD-like activity rate of the test product at all test concentrations, and Figure 2 is an excerpt of the main part of Figure 1, that is, the blending ratio of tulip extract and nicotinamide mononucleotide is 0.0001%. FIG. 3 shows the SOD-like activity rate at ~0.10%.

As shown in Figure 1, at all concentrations of tulip extract and nicotinamide mononucleotide (10%, 5%, 1%, 0.10%, 0.01%, 0.001%, 0.0001%) , Formulation 1 containing both tulip extract and nicotinamide mononucleotide had a significantly higher SOD-like activity rate than Comparative Formulation 2 and Comparative Formulation 3, each containing tulip extract and nicotinamide mononucleotide alone. it was high. From this result, it was confirmed that the combined use of tulip extract and nicotinamide mononucleotide has a synergistic effect of increasing the SOD-like activity rate.

In particular, at blended concentrations of tulip extract and nicotinamide mononucleotide shown in Figure 2 of 0.10%, 0.01%, 0.001%, and 0.0001%, the SOD-like activity rates of comparative formulations 2 and 3 were In contrast, the SOD-like activity of Formulation 1 turned positive and became significantly higher. From this result, it was confirmed that the combined use of tulip extract and nicotinamide mononucleotide has a synergistic effect that significantly increases the SOD-like activity rate at the combined concentration of both 0.0001% to 0.10%. .

Therefore, considering Experimental Example 1, the blending concentration of tulip extract and nicotinamide mononucleotide is preferably 0.0001% by weight or more, and more preferably 0.0001 to 10% by weight. Furthermore, 0.0001 to 0.10% by weight is most preferred.

According to the skin external preparation of the present invention, by containing tulip extract extracted from the flower part of tulips and nicotinamide mononucleotide, the SOD-like activity rate can be increased, and it has excellent antioxidant effect.

Furthermore, according to the skin external preparation of the present invention, by containing 0.0001% to 10% by weight of tulip extract and nicotinamide mononucleotide, the SOD-like activity rate can be increased, and the antioxidant effect can be increased. Excellent in

Furthermore, according to the skin external preparation of the present invention, since the tulip variety is Huang Komachi, the SOD-like activity rate can be significantly increased, and it has excellent antioxidant effects. Further, since the tulip extract is extracted by heating extraction or room temperature extraction, the tulip extract can be extracted by various methods without depending on temperature, resulting in excellent productivity.

According to the skin external preparation of the present invention, since the extraction solvent for tulip extract is water, 1,3-butylene glycol or ethanol, tulip extract can be easily extracted and the productivity is excellent.

FIG. 2 is a diagram showing the SOD-like activity of a lotion containing tulip extract and nicotinamide mononucleotide according to the present invention. FIG. 2 is an enlarged view of the main part of FIG. 1. FIG.

Claims (3)

Contains tulip extract extracted from tulip flowers and nicotinamide mononucleotide,
A skin external preparation, characterized in that the tulip extract and the nicotinamide mononucleotide are each contained in an amount of 0.0001% to 1% by weight based on the total amount of the external preparation . 2. The external preparation for skin according to claim 1 , wherein the tulip variety is Huang Komachi, and the tulip extract is extracted by heating extraction or room temperature extraction. The skin external preparation according to claim 1 or 2 , wherein the extraction solvent for the tulip extract is water, 1,3-butylene glycol, or ethanol.

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