The present invention relates to an agent for enhancing skin barrier function comprising a wavelength conversion substance, to a composition and product comprising the agent for enhancing skin barrier function , and to a method for enhancing skin barrier function in skin using the same.
The harm to skin caused by ultraviolet light includes adverse effects such as skin cancer, photoaging, skin spots, wrinkles and inflammation, which are also undesirable from the viewpoint of health and beauty.
Many measures are therefore being taken to protect the skin from ultraviolet light. Such measures include the use of sunscreens, the implementation of indoor spaces for avoidance of sunlight, and the use of head coverings and clothing treated to block UV rays and films designed to block UV rays.
It is an object of the present invention to provide a novel agent for enhancing skin barrier function that utilizes conversion of wavelength of ultraviolet light.
The present inventors have conducted active research with the aim of allowing ultraviolet light to be effectively utilized on skin. As a result, an agent for enhancing skin barrier function has been devised by finding that the expression of barrier function related proteins are enhanced by irradiating ultraviolet light to skin cells through wavelength conversion substance that converts ultraviolet light wavelengths.
The present application provides the present invention with the aspects set forth below.
The present invention can enhance skin barrier function in skin cells by effectively making use of ultraviolet light, and is based on finding that enhanced skin barrier function results in a desirable effect on skin. The invention provides novel uses of the aforementioned compounds that have conventionally been used primarily as dyes, pigments, ultraviolet scattering agents, ultraviolet absorbers, nutrients and antioxidants. The invention also helps to improve quality of life by providing a more positive feeling for persons who have attempted to avoid ultraviolet light as much as possible for beauty or health reasons when outdoors.
The agent for enhancing skin barrier function of the invention comprises a wavelength conversion substance as an active ingredient. A wavelength conversion substance is a substance that converts the wavelength of ultraviolet light contained in incident light and emits emission light having a wavelength longer than the wavelength of the ultraviolet light.
The ultraviolet light may include UVA, UVB and UVC. According to one embodiment, the ultraviolet light is light with a peak wavelength of 200 nm to 400 nm. The ultraviolet light may also be included in incident light such as sunlight, for example. Alternatively, the incident light may be ultraviolet light, and artificially generated ultraviolet light may be used. Ultraviolet light can have various effects on the skin. As an example, ultraviolet light is known to cause sunburns, such as sunburn and suntan, and to cause DNA damage in cells. Cellular activity is altered in ultraviolet-irradiated cells, which causes altered gene expression. As an example, UV irradiation reduces gene expression of skin barrier function-related proteins.
The emission light emitted by the wavelength conversion substance has a longer wavelength than ultraviolet light, with a peak wavelength of preferably 450 nm to 700 nm. The emission light may have one or more peaks at 450 nm, 460 nm, 470 nm, 480 nm, 490 nm, 500 nm, 510 nm, 520 nm, 530 nm, 540 nm, 550 nm, 560 nm, 570 nm, 580 nm, 590 nm, 600 nm, 610 nm, 620 nm, 630 nm, 640 nm, 650 nm, 660 nm, 670 nm, 680 nm, 690 nm or 700 nm, or in any range within these values, though without being restrictive, or it may be red light, orange light, green light or blue light. According to one embodiment, the wavelength conversion substance has its main wavelength at 450 nm to 700 nm, for example 450 - 700 nm, for light emitted upon excitation with excitation light of 200 nm to 400 nm.
Examples of wavelength conversion substances include the following components: phycobiliproteins such as allophycocyanin, C-phycocyanin, R-phycocyanin, phycoerythrocyanin, B-phycoerythrin, b-phycoerythrin, C-phycoerythrin and R-phycoerythrin; natural or synthetic components such as vitamin A, β-carotene, vitamin K, vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, niacin, lycopene, gardenia, safflower, turmeric, cochineal, perilla, red cabbage, flavonoids, carotenoids, quinoids, porphyrins, anthocyanins, polyphenols; dyes such as 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, Pyranin Conch, Blue No. 1, 2,4-diaminophenoxyethanol hydrochloride, Arizulin Purple SS, Violet No. 401, Black No. 401, Helindone Pink, Yellow No. 401, Bentizine Yellow G, Blue No. 404, Red No. 104, meta-aminophenol; and phosphors obtained by fluorescent-doping of inorganic compounds, for example, blue phosphors comprising the amorphous silica particles mentioned in Japanese Patent No. 6424656, cerium and phosphorus and/or magnesium, and red phosphors comprising compounds obtained by europium activation of mixed crystals consisting of the alkaline earth metal sulfides described in Japanese Patent No. 6361416 combined with gallium compounds, the zinc oxide phosphors mentioned in International Patent Publication No. 2018/004006, the zinc oxide phosphors mentioned in Japanese Unexamined Patent Publication No. 2018-131422, and the inorganic phosphors mentioned in Japanese Unexamined Patent Publication HEI No. 5-117127. According to one embodiment, the inorganic phosphor is one or more phosphors selected from among phosphors obtained by doping zinc oxides represented by ZnO: Zn, Zn1+z, ZnO1-x with the sulfur-containing compounds mentioned in International Patent Publication No. 2018/004006, including sulfides and/or sulfates such as zinc sulfide or zinc sulfate, magnesium titanate phosphors obtained by doping magnesium titanates such as MgTiO3 or Mg2TiO4 with manganese, and calcium phosphate phosphors obtained by doping calcium phosphates such as Ca(H2PO4)2, CaHPO4 or Ca3(PO4)2 with cerium.
The wavelength conversion substance may be obtained by extraction from a natural source such as an animal, plant or algae, or it may be obtained by an artificial method such as chemical synthesis. For example, phycobiliproteins can be prepared by extraction from algae, including blue-green algae such as spirulina (Spirulina platensis) or red algae such as porphyridiophylla (Porphyridium purpureum), by the method described in Japanese Patent No. 4048420, Japanese Patent No. 4677250 or Japanese Patent No. 3303942, for example. Zinc oxide phosphors can be produced by the method described in International Patent Publication No. 2018/004006, Japanese Unexamined Patent Publication No. 2018-131422 or Japanese Unexamined Patent Publication HEI No. 5-117127, for example. Magnesium titanate phosphors can be produced by the method described in Japanese Unexamined Patent Publication No. 2017-88719. Calcium phosphate phosphors can be produced by the method described in International Patent Publication No. 2018/117117.
So long as the wavelength conversion effect of the invention is not impaired, these wavelength conversion substances may be composed of, or may include, the components mentioned above, and they may be single components alone or combinations of more than one of the components. For example, the aforementioned phycobiliproteins or inorganic material phosphors may be mixed with other wavelength conversion substances such as B vitamins (vitamin B1, vitamin B2, vitamin B6 or vitamin B12) to exhibit synergistic effects. These components are merely examples, however, and any other substances that exhibit the wavelength conversion effect of the invention may be used.
The wavelength conversion substance content in the agent for enhancing skin barrier function, composition or product of the invention is not particularly restricted so long as the wavelength conversion effect of the invention is not impaired, and it may be appropriately determined for the type of wavelength conversion substance and the purpose of use of the agent for enhancing skin barrier function or composition. It may be any content in the range of 0.01 to 99.99 wt% or 0.1% to 999 wt%, for example.
When ultraviolet light is irradiated to the skin barrier function enhancing agent of the present invention, it emits emission light. Emission light can enhance the expression of skin barrier function-related proteins in skin cells, thereby exerting an effect for enhancing skin barrier function. Enhanced skin barrier function may be an effect that restores skin barrier function reduced by ultraviolet light, or may be an effect that enhances skin barrier function reduced by causes other than ultraviolet light. An agent for enhancing skin barrier function can also be referred to as an agent for improving skin barrier function. The agent for enhancing skin barrier function of the present invention can be used for any subject, but may be applied to a subject exposed to ultraviolet rays outside, or to a subject having a reduced skin barrier function.
Skin barrier function-related proteins are directed to proteins that can enhance the amount of any substance that enhances skin barrier function in the epidermis, or can promote the formation of structures that enhance skin barrier function. As skin barrier function-related proteins, they may be proteins that constitute the structure of the stratum corneum, or may be proteins that are involved in the production of components or the formation of structures, which are capable of enhancing skin barrier function. Skin barrier function-related proteins include at least one selected from comeodesmosine (CDSN), sphingomyelin phosphodiesterase (SMPD1), filaggrin (FLGs), involucrin (INVs), loricrin (LORs), transglutaminase 1 (TGase1), and caspase 14 (CASP14). The agent for enhancing skin barrier function of the present invention can absorb ultraviolet rays and promote the expression of the skin barrier function-related protein by emitting emission light. Accordingly, the agent for enhancing skin barrier function of the present invention may also be referred to as an expression promoting agent of a skin barrier function related protein.
Comeodesmosine (CDSN) is a cell-adhesion protein present in corneodesmosomes that adhere between keratinocytes. It has been reported that the mutation has been added to the gene of corneodesmosin in the PSD (peeling skin disease) patient, and that stratum corneum is peeled off by the malfunction of comeodesmosin. Corneodesmosine contributes to skin barrier function because it adheres between keratinocytes.
Sphingomyelin phosphodiesterase 1 (SMPD1) is an enzyme that metabolizes sphingomyelin, a type of sphingolipid. Degradation of sphingomyelin results in the formation of phosphorylcholine and ceramide. During differentiation from the granular layer to the stratum corneum, sphingomyelin is degraded by the action of SMPD1 to form ceramide. Generated ceramides contribute to skin barrier function as intercellular lipids.
Filaggrin (FLG) is a protein produced in granule cells of the epidermis. It is synthesized as a precursor profilaggrin, and during the formation of the stratum corneum, it undergoes dephosphorylation and hydrolysis to generate filaggrin. The generated filaggrin binds to keratin in the cytoplasm and aggregates keratin. Filaggrin is subsequently degraded to amino acids by degrading enzymes such as caspase 14 and functions as a natural moisturizing factor. The produced natural moisturizing factors contribute to skin barrier function.
Involculin (INV) is a protein generated in spiny cells. Involculin and loricrin, which is generated in granule cells, are major components of the peripheral zone. Involculin and loricrin are cross-linked by transglutaminase during keratinization to form insoluble structures that line the plasma membrane of keratinocites as a peripheral zone. Thus, the crosslinked involucrin imparts strength to the stratum corneum and also contributes to the skin barrier function.
Loricrin (LOR) is a protein generated in granule cells. Loricrin and involucrin are major components of the peripheral zone. Loricrin and involucrin are cross-linked by transglutaminase during keratinization to form insoluble structures that line the plasma membrane of corneocytes as a peripheral zone. Thus crosslinked loricrin provides strength to the stratum corneum and also contributes to skin barrier function.
Transglutaminase 1 (TGase1) is a protein-crosslinking enzyme. TGase 1 forms Cross-links between glutamine residues in proteins and lysine residues of heterologous or homologous proteins. The activity of transglutaminase 1 is regulated by calcium levels. During the process of differentiating granule cells into the corneocyte, cell death is occured, and the influx of calcium into the cytoplasm activities transglutaminase 1 in the corneocyte. Activated transglutaminase 1 cross-links loricrin and involucrin, thereby forming the peripheral zone. The peripheral zone lines the plasma membrane of corneocytes. This provides strength to the stratum corneum and also contributes to skin barrier function.
Caspase-14 (CASP14) is a cysteine protease. It is expressed as pro-caspase 14 in granule cells from spiny cells and is activated in the stratum corneum. Active caspase 14 degrades part of filaggrin. The action of additional enzymes on degraded filaggrin produces natural moisturizing factors (NMFs), which contribute to skin barrier function.
Any form of administration may be used for the agent for enhancing skin barrier function and composition of the invention, but an external preparation for skin will often be preferred, such as a drug, quasi drug or cosmetic, for enhancing skin barrier function by exposing the skin to light containing ultraviolet light. When the agent for enhancing skin barrier function or composition of the invention is to be used as an external preparation for skin, the dosage form, coating method and number of doses may be determined as desired. For example, it may be applied onto skin in the form of cosmetic water or a spray, oil, cream, latex, gel, sunscreen or suntan lotion either periodically or irregularly, once or several times per day at morning, noon or evening, or before going out or engaging in outdoor activities, marine sports or skiing, for example, when exposure to sunlight is expected.
The agent for enhancing skin barrier function and composition of the invention may also be used in combination with an additive such as an excipient, preservative, thickener, binder, disintegrator, dispersing agent, stabilizer, gelling agent, antioxidant, surfactant, preservative, oil, powder, water, alcohol, thickener, chelating agent, silicone, antioxidant, humectant, aromatic, drug component, antiseptic agent, pH adjustor or neutralizer, selected as necessary or desired. It may also be used in combination with other agent for enhancing skin barrier function to increase the effect of the invention.
The present invention further provides products such as sun visors, caps, clothing, gloves, screen films, window sprays or creams, window materials or wall materials, for example, that comprise the agent for enhancing skin barrier function of the invention and are intended to enhance skin barrier function by exposing the skin to light containing ultraviolet light. The usage of additives in the products of the invention and the forms of the products may also be as desired.
The present invention further provides a method for producing the agent for enhancing skin barrier function, composition or product of the invention. A method for enhancing skin barrier function in the skin of a subject is also provided, the method comprising application of the agent for enhancing skin barrier function or composition of the invention onto the skin of a subject and exposing the skin to light containing ultraviolet light after application of the agent for enhancing skin barrier function or composition; or passing light containing ultraviolet light through the product of the invention, and exposing the skin to the transmitted light; wherein the agent for enhancing skin barrier function, composition or product converts the wavelength of ultraviolet light in the incident light and emits emission light with a longer wavelength than the wavelength of the ultraviolet light, transmitting the ultraviolet light with a peak wavelength of preferably 200 nm to 400 nm as light with a peak wavelength of 450 nm to 700 nm, for example 500 nm to 700 nm. The method for enhancing skin barrier function in skin of an subject will often be for the purpose of beautifying, instead of treatment by a doctor or medical worker.. The invention further provides a cosmetic counseling method for supporting cosmetology, which includes providing a cosmetic method, an agent for enhancing skin barrier function, composition or product of the invention to a subject.
The present invention will now be explained in greater detail by examples. However, the invention is in no way limited by the examples.
Experiment 1: Change in gene expression by applying wavelength conversion substances Experiment 1-1: Preparation of wavelength conversion substances Wavelength conversion substances were prepared in the following manner.
Cell samples were prepared in the following manner.
Experiment 2-1: RNA extraction Cell samples incubated for 24 h after irradiating ultraviolet light in Experiments 1-3 were washed with 500 µl warm PBS, and PBS was completely aspirated. Qiagen RNeasy Mini Kit prep (Qiagen,74106) were used to extract RNA according to the product instructions.
Microarray for Human gene-expression (SurePrint G3 Human GE Microarray 8x60K Ver. 3.0 (Agilent technology)) was used to perform the analysis of RNAs extracted in Experiment 2-1. The extracted RNA was subjected to labeling reactions, amplification reactions, purification, and quantitation of cRNA according to the protocol provided by Agilent Technology to prepare hybridization samples.. Microarrays were observed with AGILINT C MICROARRAY SCANNER to identify genes whose expression was significantly reduced or increased by the presence or absence of wavelength-converting material, and the results are shown below.
Experiment 3-1: RNA extraction
1. SuperScript VILO cDNA Synthesis Kit (Thermo Fisher) was used in accordance with the product instructions, with 1 pg to 2.5 µg of RNA being added per container, and the PCR system was operated at a setting of at 25° C. for 10 min, at 42° C. for 60 min, at 85° C. for 5 min, and preservation at 4° C.
The reverse-transcripted sample was diluted 50-fold with RNase free water, and a further 5-fold dilution series was prepared. Then, the reaction system described below was prepared and measured by a real-time PCR instrument (Applied Biosystems). ΔCt was determined based on the Ct value for each gene and the Ct value of the internal standard, GAPDH in the test sample (FLGs, INVs, and CDSN). In addition, ΔΔCt was determined for Sham sample (UV-unirradiated sample) and calculated as relative amount 2-AACt (SMPD1 and TGase1) (
These results demonstrates that the expression of skin barrier function-related proteins was promoted by irradiating UV to wavelength-converting materials. Thereby, an effect of enhancing skin barrier function was exerted.
The embodiments of the invention described above are not intended to place limitations on the invention, and various modifications including cosmetics and drug compositions may be incorporated, which fall within the gist of the invention.
Number | Date | Country | Kind |
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2020-015641 | Jan 2020 | JP | national |
Filing Document | Filing Date | Country | Kind |
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PCT/JP2021/003403 | 1/29/2021 | WO |