JP2024031642A - A beauty method to enhance the physiological effects of riboflavin using visible light - Google Patents

Abstract

An object of the present invention is to obtain a component whose medicinal efficacy is increased by irradiation with visible light. Ultraviolet light promotes the expression of matrix proteinases and/or inflammatory cytokines. Riboflavin exerts a physiological effect that suppresses this enhanced expression. On the other hand, it has been found that by being exposed to visible light, the physiological effects of riboflavin are enhanced and the expression of matrix proteinase and/or inflammatory cytokines is further suppressed. As a result, we have developed a cosmetic method for enhancing the physiological effects of riboflavin, which includes applying a topical skin preparation containing riboflavin and irradiating it with visible light, a wavelength converting substance that converts ultraviolet rays into visible light, and riboflavin. The present invention provides a beauty kit, which includes a skin external composition containing, a skin external preparation containing riboflavin, and a beauty irradiator that irradiates visible light of 1000 to 10000 Lux at a wavelength in the visible light range. [Selection diagram] Figure 1A-B

Description

The present invention relates to a cosmetic method for enhancing the physiological action of riboflavin, which includes applying a topical skin preparation containing riboflavin and exposing it to visible light, and a method for enhancing the physiological effects of riboflavin, which includes riboflavin and a wavelength converting substance that converts ultraviolet rays into visible light. The present invention also relates to a beauty kit comprising a composition for external use on the skin, an external preparation for skin containing riboflavin, and an irradiator that irradiates visible light.

Natural light is mainly classified into ultraviolet light, visible light, and infrared light. Ultraviolet rays are known to have adverse effects on the skin, such as skin cancer, photoaging, age spots, wrinkles, and inflammation, and are undesirable from the viewpoint of health and beauty. Many measures have been taken to protect the skin from ultraviolet radiation. Examples include using sunscreen, staying indoors to avoid exposure to sunlight, using hats and clothing with UV protection, and using UV protection film. On the other hand, it has been reported that visible light has the effect of activating cells, and beauty methods and beauty devices that irradiate visible light with a specific wavelength have been developed. For example, it has been reported that red light of about 630 nm reaches deep into the skin, improves the function of macrophages, and activates dermal cells. Yellow and green light with shorter wavelengths activate epidermal cells, and are expected to have effects such as suppressing melanin production and improving stratum corneum turnover. With the aim of utilizing the cellular effects of visible light while avoiding ultraviolet rays, the development of beauty methods and cosmetics that utilize wavelength conversion substances that absorb the wavelength of irradiated ultraviolet rays and emit light with longer wavelengths has been developed. It is progressing.

Patent No. 6424656 Patent No. 6361416 International Publication No. 2018/004006 Japanese Patent Application Publication No. 2018-131422 Japanese Patent Application Publication No. 5-117127 Patent No. 4048420 Patent No. 4677250 Patent No. 3303942 Japanese Patent Application Publication No. 2017-88719 International Publication No. 2018/117117

PLoS One. 2016 Sep 20;11(9):e0161580. doi: 10.1371

An object of the present invention is to obtain a component whose medicinal efficacy is increased by visible light.

The present inventors conducted a screening to find a component that increases its medicinal efficacy when exposed to visible light, and found that riboflavin, a type of water-soluble vitamin, increased its medicinal efficacy when exposed to visible light. It has been found to be a scalable component. More specifically, riboflavin has the medicinal effect of suppressing the expression of inflammatory cytokines and matrix proteinase, and can suppress the expression promoted by ultraviolet rays. It was discovered that irradiation further suppresses the expression of inflammatory cytokines and matrix proteinase, leading to the present invention.
Therefore, the present application provides the following invention.
[1] A cosmetic method for enhancing the physiological action of riboflavin, which comprises applying a topical skin preparation containing riboflavin and exposing the skin to visible light.
[2] The beauty method according to item 1, which suppresses rough skin, wrinkles, or age spots, or brings about whitening or anti-aging by enhancing the physiological action of riboflavin.
[3] The cosmetic method according to item 1, wherein the physiological action of riboflavin is to suppress the expression of matrix proteinase or inflammatory cytokines in keratinocytes.
[4] A skin external composition containing a wavelength converting agent that converts ultraviolet rays into visible light and riboflavin.
[5] The skin external composition according to item 4, wherein the wavelength converting agent converts the wavelength from ultraviolet rays to visible light, and the visible light enhances the physiological action of riboflavin.
[6] The composition for external use on skin according to item 5, wherein the wavelength converting agent converts ultraviolet rays into visible light having a peak wavelength at 450 to 700 nm, 450 to 600 nm, or 500 to 550 nm.
[7] The skin external composition according to item 5, wherein the wavelength converting agent is a zinc oxide phosphor.
[8] A beauty kit comprising a skin external preparation containing riboflavin and a beauty irradiator that irradiates visible light of 1000 to 10000 Lux at a wavelength in the visible light range.

In the present invention, the physiological action of riboflavin is enhanced by applying a skin external preparation containing riboflavin to the skin and then exposing it to visible light.

Figure 1 shows that the expression of inflammatory cytokines ((A): IL-8, (B) TNF-α), which was increased by ultraviolet irradiation, was suppressed by the action of riboflavin, and further visible light irradiation suppressed the expression of inflammatory cytokines ((A): IL-8, (B) TNF-α). This is a graph showing that. Figure 1 shows that the expression of matrix proteinases ((C) MMP9, (D) MMP1), which was enhanced by ultraviolet irradiation, was suppressed by the action of riboflavin, and further irradiation with visible light enhanced the action of riboflavin. It is a graph.

The present invention relates to a cosmetic method for enhancing the physiological action of riboflavin, which includes applying a topical skin preparation containing riboflavin and exposing the skin to visible light.

In the present invention, the physiological action of riboflavin is enhanced by exposing the skin to which the external skin preparation containing riboflavin has been applied to visible light. The visible light may be directly irradiated using an irradiator, or may be irradiated indirectly by light emitted through a wavelength converting substance. In addition to visible light irradiation, ultraviolet rays may be irradiated. Ultraviolet radiation can cause harmful physiological effects on the skin. Examples include increasing reactive oxygen species, activating matrix proteinases, and inducing DNA damage. Furthermore, ultraviolet rays enhance the expression of matrix proteinases and/or inflammatory cytokines, and riboflavin exerts a physiological effect of suppressing the expression thus enhanced. On the other hand, by being exposed to visible light, the physiological effects of riboflavin are enhanced, and the expression of matrix proteinase and/or inflammatory cytokines is further suppressed (FIG. 1). Being exposed to visible light may mean exposure to electric lights or sunlight in daily life, or irradiation with visible light using an irradiator. When exposed to sunlight, it is preferable to use a wavelength converting substance and/or an ultraviolet absorbing substance in order to soften the effects of ultraviolet rays contained in sunlight. The beauty method of the present invention can be performed, for example, in a beauty salon or the like by a beauty practitioner other than a doctor using an irradiator capable of emitting visible light of a specific wavelength.

The visible light irradiated in the present invention is a visible light having a peak wavelength of 450 to 700 nm, particularly from the viewpoint of promoting the physiological action of flavonoids, and the peak of the visible light is further 450 to 600 nm, particularly 500 to 550 nm. Preferably present. From the viewpoint of irradiating red light, the peak range of visible light is preferably 600 to 700 nm, from the viewpoint of irradiating yellow light, the peak range of visible light is preferably 550 to 600 nm, and from the viewpoint of irradiating green light, the peak range of visible light is preferably 600 to 700 nm. is preferably 500 to 550 nm. The intensity of visible light can be selected as appropriate, and can be appropriately selected between 1000 and 10000 Lux. Such visible light wavelengths and intensities can be applied using illuminators well known in the art. Preferably, a single peak is present in a given peak range, but two or more peaks may be present. Overlapping peaks may make it difficult to distinguish between peaks. From the viewpoint of irradiating visible light with a single peak, it is preferable to use an LED as the irradiator. Furthermore, the peak range can be narrowed down by using a film or the like for the light emitter.

The external skin preparation of the present invention may be any external skin preparation as long as it contains riboflavin, or may be an external skin preparation containing a riboflavin-containing extract. Such external skin preparations may be pharmaceuticals, quasi-drugs, cosmetics, and the like. The dosage form, application method, number of administrations, etc. of the skin external preparation used in the cosmetic method of the present invention can be arbitrarily determined. For example, it may be in the form of a lotion, spray, oil, cream, emulsion, gel, sunscreen, or suntan agent. Before visible light irradiation in the cosmetic method of the present invention, the skin external preparation may be applied once or multiple times, and then visible light may be irradiated immediately or after a certain period of time, for example, 10 minutes to several hours. Instead of being irradiated with visible light, visible light may be irradiated when the subject is active outdoors or indoors with lights on.

Riboflavin is also known as vitamin B2, and is known to contribute to the metabolism of fats, carbohydrates, and proteins, respiration, and the formation of red blood cells in vivo. Riboflavin is essential for maintaining the health of the skin, nails, and hair, and a deficiency causes symptoms such as stomatitis, dermatitis, keratitis, hair loss, and hyperemia. It can be added to cosmetics and medicines to maintain normal skin function. As an effect of riboflavin on the skin, it has been shown to suppress the mRNA expression of UV-induced inflammatory cytokines (IL-1α, GM-CSF) (Non-Patent Document 1: PLoS One. 2016 Sep 20;11(9): e0161580. doi: 10.1371). Further, in the same literature, as an effect of riboflavin on the skin, it was shown that UV-induced MMP1 mRNA expression was suppressed, but the protein amount of MMP1 was not changed.

Inflammatory cytokines are cytokines released by cells that undergo physical, chemical, or biological stimulation and induce inflammation. When inflammatory cytokines are released, they induce the accumulation of immune cells at the site of inflammatory cytokine release and promote activation and proliferation of immune cells. Inflammatory cytokines include IL-8, IL-6, IL-1β, IL-1α, IL-10, INF-γ, TNF-α, and GM-CSF. Among these, IL-8 and TNF-α are preferred as inflammatory cytokines from the viewpoint of being expressed in keratinocytes. Abnormally enhanced expression of inflammatory cytokines can cause rough skin, inflammatory diseases such as atopic dermatitis, and psoriasis, and can also activate melanocytes and cause changes in skin tone. Therefore, by suppressing the expression of inflammatory cytokines, rough skin and age spots can be prevented, alleviated, treated, or suppressed, and anti-aging and whitening effects can be exerted.

The matrix proteinase may be any matrix proteinase belonging to the matrix metalloproteinase family, but MMP1, MMP2, MMP9, and/or MMP13 are particularly preferred from the viewpoint of expression in the skin. Matrix proteinases degrade extracellular matrices consisting of collagen, proteoglycans, elastin, etc. Ultraviolet irradiation increases the expression of matrix proteinase in the skin, particularly in the epidermis and/or dermis, degrading the extracellular matrix and causing wrinkles and sagging. In addition, decomposition products can serve as signals and cause various physiological phenomena. Therefore, by suppressing the expression of matrix proteinase, wrinkles can be prevented, reduced, treated, or suppressed, or anti-aging effects can be exerted.

Physiological effects of riboflavin on the skin, particularly on epidermal cells, include suppressing the expression of inflammatory cytokines and/or matrix proteinases. Without intending to be limited by theory, it is believed that riboflavin suppresses inflammatory cytokine and/or matrix proteinase expression in keratinocytes (Figure 1), thereby inhibiting wrinkles or age spots, or whitening or anti-aging. can bring about

In order to exert the physiological effects exerted by riboflavin, a skin external preparation containing an extract containing riboflavin may be used instead of riboflavin. As such an extract, riboflavin, particularly an extract obtained from a raw material containing riboflavin can be used, and examples include yeast extract, mushroom extract, meat extract, fish extract, cod liver oil, and the like.

The extract described in this specification can be obtained by a conventional method, for example, by immersing the plant of origin with an extraction solvent at room temperature or heating, or heating and refluxing, filtering, and concentrating. As the extraction solvent, any solvent commonly used for extraction can be used, such as an aqueous solvent such as water, physiological saline, a phosphate buffer, a borate buffer, or an organic solvent such as ethanol, Alcohols such as propylene glycol, 1,3-butylene glycol, and glycerin, hydrous alcohols, chloroform, dichloroethane, carbon tetrachloride, acetone, ethyl acetate, hexane, and the like can be used alone or in combination. Preferably, a mixed solvent of water and alcohol, such as 1,3-butylene glycol, is used as the solvent. The extract obtained by extraction with the above solvent can be used as it is, or an extract concentrated by freeze-drying can be used. If necessary, an extract with impurities removed using an adsorption method, such as an ion exchange resin, or a porous extract can be used. It is also possible to use a product that is adsorbed on a column of polymer (for example, Amberlite XAD-2), eluted with a desired solvent, and further concentrated.

In the present invention, a wavelength conversion substance refers to a substance that absorbs incident light of a specific wavelength and emits output light of a longer wavelength than the incident light. The incident light absorbed by the wavelength conversion substance of the present invention is preferably ultraviolet light. Ultraviolet light may include UVA, UVB, UVC, and the like. In some embodiments, the ultraviolet light is light with a peak wavelength between 200 nm and 400 nm. Furthermore, incident light such as sunlight may include ultraviolet rays. Alternatively, the incident light may be ultraviolet light, or artificially generated ultraviolet light may be used.

The output light emitted by the wavelength conversion substance has a longer wavelength than ultraviolet light, preferably having a peak wavelength in the range of 450 nm to 700 nm, such as 500 nm to 700 nm or 450 to 600 nm, more preferably 500 to 550 nm. The emitted light is, for example, but not limited to, 450nm, 460nm, 470nm, 480nm, 490nm, 500nm, 510nm, 520nm, 530nm, 540nm, 550nm, 560nm, 570nm, 580nm, 590nm, 600nm, 610nm, 620nm, 630nm. nm, 640 nm, It may have one or more peaks at 650nm, 660nm, 670nm, 680nm, 690nm, 700nm, or any range of these values, or it may have red light, orange light, green light, blue light, etc. There may be. In one embodiment, the wavelength converting material exhibits a dominant wavelength of light emitted when excited with excitation light of 200 nm to 400 nm, such as 500 nm to 700 nm or 450 to 600 nm, more preferably 500 to 550 nm.

Examples of wavelength converting substances include the following components: allophycocyanin, C-phycocyanin, R-phycocyanin, phycoerythrocyanin, B-phycoerythrin, b-phycoerythrin, C-phycoerythrin, R-phycocyanin. Phycobiliproteins such as coerythrin; vitamin A, β-carotene, vitamin K, vitamin B1, vitamin B6, vitamin B12, folic acid, niacin, lycopene, gardenia, safflower, turmeric, cochineal, perilla, red cabbage, flavonoids, carotenoids, quinoids, Naturally derived or synthetic ingredients such as porphyrins, anthocyanins, polyphenols; Red No. 401, Red No. 227, Red No. 504, Red No. 218, Orange No. 205 P, Yellow No. 4, Yellow No. 5, Green No. 201, Pyranine Conc. , Blue No. 1, 2,4-diaminophenoxyethanol hydrochloride, Alizurin Purple SS, Purple No. 401, Black No. 401, Herringdon Pink, Yellow No. 401, Benzidine Yellow G, Blue No. 404, Red No. 104, Meta-aminophenol dyes such as; phosphors doped with inorganic compounds to give fluorescence; for example, blue phosphors containing amorphous silica particles, cerium, phosphorus and/or magnesium described in Patent No. 6424656; A red phosphor containing a compound in which europium is activated in a mixed crystal of an alkaline earth metal sulfide and a gallium compound described in No. 6361416, a zinc oxide phosphor described in International Publication No. 2018/004006, and JP-A-2018- Examples include the zinc oxide phosphor described in No. 131422; the inorganic phosphor described in JP-A-5-117127; and the like. In some embodiments, the inorganic phosphor comprises zinc oxide, which can be represented as ZnO:Zn, Zn _1+z , ZnO _1-x, as described in WO 2018/004006, e.g., zinc sulfide, zinc sulfate. phosphors doped with sulfur-containing compounds such as sulfides and/or sulfates, magnesium titanate phosphors doped with manganese, and magnesium titanate phosphors doped with manganese, such as MgTiO ₃ , Mg ₂ TiO ₄ , and Ca(H ₂ PO ₄ ) ₂ , CaHPO ₄ , Ca ₃ (PO ₄ ) ₂ , which are calcium phosphate phosphors doped with cerium. In another example, zinc oxide phosphor is obtained by reducing and firing zinc oxide, has oxygen defects, and is thought to have an average compositional formula of Zn _1+z O or ZnO _1-x . This is what is being done.

The wavelength conversion substance may be obtained by a method such as extraction from natural products such as animals, plants, and algae, or may be obtained by an artificial method such as chemical synthesis. For example, phycobiliproteins can be obtained by using algae such as blue-green algae such as Spirulina platensis and red algae such as Porphyridium purpureum by the method described in Patent No. 4048420, Patent No. 4677250, Patent No. 3303942, etc. It may also be prepared by extraction. The zinc oxide phosphor may be produced, for example, by the method described in International Publication No. 2018/004006, Japanese Patent Application Publication No. 2018-131422, and Japanese Patent Application Publication No. 5-117127. The magnesium titanate phosphor may be manufactured by the method described in JP-A-2017-88719. The calcium phosphate phosphor may be manufactured by the method described in International Publication No. 2018/117117. Among these, zinc oxide phosphors are preferred from the viewpoint that the wavelength after conversion is 500 to 550 nm.

These wavelength conversion substances may be composed of or contained in the components exemplified above, and may be used alone or in combination of multiple types, as long as the wavelength conversion effect of the present invention is not impaired. good. For example, a synergistic effect may be achieved by mixing other wavelength converting substances such as vitamin B (vitamin B1, vitamin B6, vitamin B12, etc.) with the phycobiliprotein or inorganic phosphor. However, these components are just examples, and any substance that exhibits the wavelength conversion effect of the present invention can be used.

Another aspect of the present invention relates to a skin external composition comprising a wavelength converting substance that converts ultraviolet rays into visible light and riboflavin. A wavelength converting substance is blended into such a composition for external use on the skin as a physiological action enhancer of riboflavin. As a result, when the skin external composition is applied to the skin and placed under light irradiation, the wavelength conversion substance converts the wavelength from ultraviolet rays to visible light, and the visible light enhances the physiological action of riboflavin. can. By enhancing the physiological effects of riboflavin, it particularly enhances the inhibitory effect on inflammatory cytokine and/or matrix proteinase expression, thereby causing inflammation such as rough skin, wrinkles, spots, sagging, skin aging (particularly photoaging), erythema, and sunburn. can exert the effect of suppressing Therefore, the skin external composition of the present invention may be an antioxidant enzyme expression promoter, an agent for suppressing rough skin, wrinkles, spots, sagging, skin aging, an agent for suppressing inflammation such as erythema and sunburn, or an agent for whitening or It may also be an anti-aging agent. The composition for external use on the skin according to the present invention is expected to be exposed to sunlight regularly or irregularly, for example, once to several times a day, such as in the morning, afternoon, and evening, while going out, doing outdoor activities, marine sports, skiing, etc. It may be applied to the skin each time, such as before being treated.

In addition, the external skin preparation or skin external composition of the present invention may contain, as necessary, excipients, preservatives, thickeners, binders, disintegrants, dispersants, stabilizers, gelling agents, Antioxidants, surfactants, preservatives, oils, powders, water, alcohols, thickeners, chelating agents, silicones, antioxidants, humectants, fragrances, various medicinal ingredients, preservatives, pH adjusters, Additives such as neutralizing agents can be arbitrarily selected and used in combination. Furthermore, in order to enhance the effects of the present invention, other cell activators and the like may be used in combination.

In a further aspect of the present invention, the present invention provides a topical skin preparation containing riboflavin and irradiation with visible light of 1000 to 10000 Lux at a wavelength in the visible light range, particularly 450 to 700 nm, 450 to 600 nm, or 500 to 550 nm. The present invention may also relate to a beauty kit, which includes a irradiator for irradiation. Such a beauty kit may be used by a purchaser or a beauty practitioner. The irradiator that irradiates visible light can be irradiated by using an irradiator that is well known in this technical field. From the viewpoint of irradiating visible light in a predetermined peak range, it is preferable to use an LED.

Next, the present invention will be explained in more detail with reference to Examples. Note that the present invention is not limited to this.

Experiment 1: Cell culture 1. Normal human epidermal keratinocytes (KK-4109, Kurabo Industries, Ltd.) were cultured using HuMedia-KG2 medium (KK-2150S, Kurabo Industries, Ltd.).
2. After the cells reached approximately 50% confluence, the medium was replaced with EpiLife medium (MEPI500CA ThermoFisher).
3. After 24 hours, riboflavin (R4500 Sigma-Ardrich) was added to the riboflavin addition group at a final concentration of 5 μM.
4. 24 hours after addition of riboflavin, the medium was replaced with Hanks' Balanced Salt Solution (HBSS) prior to light irradiation.
5. The ultraviolet (UVB) irradiation group was irradiated with 10 mJ/cm ² using a TL20W/12RS lamp (PHILIPS) and a UG11 filter (SCHOTT).
6. For the group to be irradiated with both ultraviolet and visible light, after ultraviolet irradiation, visible light was irradiated using a solar simulator (91192-1000 Newport) and KG1 and GG420 filters (SCHOTT) (integrated irradiation from wavelength 400 nm to 800 nm). The amount is 50 J/cm ² ).
7. After completion of light irradiation, the medium was exchanged from HBSS to EpiLife medium, and the cells were cultured for an additional 24 hours. Riboflavin was added to the riboflavin-added group at a final concentration of 5 μM.

Experiment 2: RT-PCR
1. Immediately after the completion of Experiment 1, RNA was extracted using the RNeasy Mini Kit (QIAGEN) according to the product instructions.
2. RNA samples extracted from each specimen were subjected to RT-PCR reaction using QuantiTect SYBR Green RT-PCR Kit (204243, QIAGEN) and Light Cycler (Roche) according to the product instructions, and IL8, TNFα, MMP1, MMP9 and The amount of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was quantified as an internal standard. The sequences of the primers used in the experiment are shown below.


3. The relative expression levels of IL8, TNFα, MMP1, and MMP9 with respect to the GAPDH expression level of each sample were determined and compared.

The embodiments of the present invention have been described above. However, the present invention is not limited to these, and may be modified as appropriate to cosmetics, pharmaceutical compositions, etc. without departing from the spirit of the invention.

Claims (8)

A cosmetic method for enhancing the physiological action of riboflavin, which comprises applying a topical skin preparation containing riboflavin and exposing it to visible light. The cosmetic method according to claim 1, which suppresses rough skin, wrinkles, or age spots, or brings about whitening or anti-aging by enhancing the physiological action of riboflavin. The cosmetic method according to claim 1, wherein the physiological effect of riboflavin is to suppress the expression of matrix proteinase or inflammatory cytokines in keratinocytes. A skin external composition comprising a wavelength converting agent that converts ultraviolet rays into visible light and riboflavin. The composition for external use on skin according to claim 4, wherein the wavelength converting agent converts the wavelength from ultraviolet light to visible light, and the visible light enhances the physiological action of riboflavin. The composition for external use on skin according to claim 5, wherein the wavelength converting agent converts ultraviolet light into visible light having a peak wavelength of 450 to 700 nm. The skin external composition according to claim 5, wherein the wavelength converting agent is a zinc oxide phosphor. A beauty kit that includes a skin external preparation containing riboflavin and a beauty irradiator that irradiates visible light of 1000 to 10000 Lux at a wavelength in the visible light range.

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