Ophthalmic components

Abstract

To remove senescent cells, and / or to prevent, suppress, or improve eye aging by removing senescent cells. [Solution] An ophthalmic composition containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and lotus.

Description

[Technical Field]

[0001] The present invention relates to an ophthalmic composition. [Background Art]

[0002] In recent years, cellular senescence, a phenomenon in which cell proliferation is permanently arrested, has attracted attention. For example, in an analysis using genetically modified mice, p16 involved in the induction of cellular senescence INK4α It has been reported that elimination of cells expressing the gene reduces the incidence of various age-related diseases such as atherosclerosis, cataract, sarcopenia, and carcinogenesis (Non-Patent Document 1).

[0003] Cells in which cell proliferation is permanently arrested are referred to as senescent cells. It is known that senescent cells do not merely stop proliferating, but also produce and secrete various physiologically active substances such as proteases including extracellular matrix degrading enzymes, inflammatory cytokines, chemokines, growth factors, and exosomes. This phenomenon is known as "senescence-associated secretory phenotype (SASP)" (Non-Patent Document 1). In the present invention, the physiologically active substance secreted by SASP may sometimes be referred to as a SASP factor.

[0004] It has been reported that SASP factors secreted from senescent cells act on neighboring cells to induce inflammatory responses, promote or suppress cell proliferation, induce cellular senescence, and the like. That is, senescent cells can, through secretion of SASP factors, trigger an inflammatory response in neighboring normal (non-senescent) cells, or convert normal cells into senescent cells. Therefore, it is expected that elimination of senescent cells that secrete SASP factors will suppress inflammatory responses, cellular senescence, and the like in surrounding cells.

[0005] Patent Document 1 discloses that elimination of senescent cells can treat or prevent meibomian gland dysfunction.

[0006] Furthermore, as shown in Non-Patent Document 3, senescent cells are known to be SA-β-gal (senescence-associated beta galactsidase) positive. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Eiji Hara, "A New Phase in Cellular Senescence: Potential for a More Complete Understanding and Extending Healthy Lifespan," Experimental Medicine, July 2019, Vol. 37, No. 11, pp. 1728-1734. [Non-Patent Document 2] Waaijer ME et al. "P16INK4a Positive Cells in Human Skin Are Indicative of Local Elastic Fiber Morphology, Facial Wrinkling, and Perceived Age." J Gerontol A Biol Sci Med Sci. 2016 Aug;71(8):1022-1028. [Non-Patent Document 3] Dimri GP et al., "A biomarker that identifies senescent human cells in culture and in aging skin in vivo." Proc Natl Acad Sci US A. 1995 Sep 26;92(20):9363-9367.

[0008] [Patent Document 1] Japanese Patent Publication No. 2024-125057 [Overview of the project] [Problems that the invention aims to solve]

[0009] As mentioned above, p16 INK4αGenes are known to be involved in inducing cellular senescence, and a correlation has been suggested between the number of p16INK4a-positive cells in ocular mucosal tissue and the aging phenomenon of the eye, suggesting a possible link between cellular senescence and the aging phenomenon of the eye. Furthermore, as mentioned above, SASP factors secreted from senescent cells have been reported to act on neighboring cells, causing induction of inflammatory responses and cellular senescence, so it can be inferred that the accumulation of senescent cells due to aging accelerates the aforementioned aging phenomenon of the eye.

[0010] Therefore, in order to suppress the induction of inflammatory responses and cellular senescence in surrounding cells, and to prevent the progression of age-related phenomena related to the eyes, such as dry eyes, cataracts, chronic conjunctivitis, blurred vision, eye strain, and eye discomfort, it is desirable to remove as much of the aging process as possible from around the orbit.

[0011] In view of the above circumstances, the inventors of the present invention have made the invention of removing senescent cells and / or removing senescent cells The main objective was to prevent, suppress, and improve the aging process of the eyes. [Means for solving the problem]

[0012] The inventors have identified wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, and tachibana as the primary plants for this purpose. Chymbidium, sage, perilla, cherry blossom, coffee tree, peppermint, rosehip, tolmae It contains at least one plant extract selected from the group consisting of Centilla and Astragalus sinicus. We discovered that the composition has the effect of removing senescent cells, and through further improvements, we completed the present invention. I reached it.

[0013] In other words, the present invention encompasses the following inventions. Section 1. An ophthalmic composition containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus. Section 2. An anti-aging ophthalmic composition containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus. Section 3. An ophthalmic composition for improving dry eye according to claim 1, comprising at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus. Section 4. An ophthalmic composition for improving blurred vision according to claim 1, comprising at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus. Section 5. An ophthalmic composition for improving eye strain as described in item 1, comprising at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus. [Effects of the Invention]

[0014] According to the technology disclosed herein, senescent cells can be removed, and age-related eye phenomena such as dry eyes, cataracts, chronic conjunctivitis, blurred vision, eye strain, and eye discomfort can be prevented, suppressed, or improved. [Brief explanation of the drawing]

[0015] [Figure 1] Among the results of Test Example 2, the results for wild thyme extract-1, wild thyme extract-2, rose extract, mayonnaise extract, chamomile extract, loquat extract, eucalyptus extract, common thyme extract, and sage extract are shown. The upper end of the bar graph indicates the average cell viability, and the error bars indicate ± standard deviation. [Figure 2] Among the results of Test Example 2, the results for perilla extract-1, perilla extract-2, cherry blossom extract, coffee tree extract, peppermint extract, dog rose extract, tormentilla extract, and milk-vetch extract are shown. The upper end of the bar graph indicates the average cell viability, and the error bars indicate ± standard deviation. Mode for Carrying Out the Invention

[0016] Hereinafter, each embodiment encompassed by the present invention will be described in further detail.

[0017] In the present invention, "senescent cell elimination" refers to inhibiting or suppressing the survival of senescent cells. Above all, while the composition of the present disclosure has a senescent cell elimination effect, it preferably does not inhibit the proliferation of normal cells, and particularly preferably has an effect of promoting the proliferation of normal cells. In the present invention, "normal cells" refers to SA-β-gal negative cells.

[0018] As described above, senescent cells can induce inflammatory reactions in neighboring normal cells or transform normal cells into senescent cells via secretion of SASP factors. Therefore, it is expected that the composition of the present disclosure having a senescent cell elimination effect will suppress inflammatory reactions, cellular senescence, and the like caused by SASP factors. That is, the composition of the present invention can also exert an anti-ocular mucosa senescence effect.

[0019] In the present invention, "ocular mucosa" refers to the bulbar conjunctiva and the palpebral conjunctiva.

[0020] In this invention, the term "aging of the ocular mucosa" refers to changes in the ocular mucosa associated with aging. Specifically, age-related changes in the ocular mucosa include a decrease in the number of mucous cells, a decrease in tear secretion, and chronic conjunctivitis.

[0021] In this invention, the term "anti-ocular aging" encompasses preventing, suppressing, and improving the aging of the ocular mucosa.

[0022] Furthermore, the composition of the present invention can be applied to, but is not limited to, people who want to prevent eye aging, people who want to suppress eye aging, people who want to improve eye aging, people who want to prevent a decrease in the number of mucus cells, people who want to prevent dry eye, people who want to prevent chronic conjunctivitis, people who want to prevent blurred vision, people who want to prevent eye fatigue, people who want to prevent eye discomfort, people who want to improve a decrease in the number of mucus cells, people who want to improve dry eye, people who want to improve chronic conjunctivitis, people who want to improve blurred vision, people who want to improve eye fatigue, and people who want to improve eye discomfort.

[0023] Preparation of plant extracts according to the present invention In the present invention, the term "plant extract" preferably includes any of the following: an extract obtained by an extraction operation using plants, a diluted extract, a concentrated extract, and a dried product obtained by drying the extract. Furthermore, the extract may be fractionated and / or purified by conventional methods to the extent that the desired effect can be obtained. That is, the term "plant extract" in the present invention also includes the fractions, crude products, and purified products of the extract.

[0024] The plant extract of the present invention is not particularly limited as long as the desired effect is obtained. Examples of extractable parts include roots, stems, leaves, flowers, buds, rhizomes, seeds, fruits, pericarps, branches, trunks, bark, etc. The extractable part may be a single part, a mixture of multiple parts, or the entire plant. The extractable part may be appropriately selected according to the characteristics of the plant.

[0025] The method for preparing plant extracts according to the present invention is not particularly limited as long as the desired effect is obtained. For example, the extraction solvent, the amount of extraction solvent used, the extraction time, the extraction temperature, etc., can be appropriately selected depending on the type and part of the plant used in the extraction operation. Furthermore, the plant used in the extraction operation may be dried or not, and may be shredded or pulverized.

[0026] Examples of extraction solvents include water, methanol, ethanol, propanol, isopropanol, 1,3-butylene glycol, propylene glycol, glycerin, benzene, toluene, xylene, ethyl ether, methyl ethyl ketone, methyl isobutyl ketone, dichloromethane, dichloroethane, chloroform, ethyl acetate, propyl acetate, butyl acetate, and acetone. A mixture of two or more solvents may also be used as the extraction solvent. In this case, the mixing ratio can be appropriately adjusted depending on the characteristics and condition of the plant being subjected to the extraction operation. Preferably, the extraction solvent is one or more selected from the group consisting of water, ethanol, 1,3-butylene glycol, propylene glycol, and glycerin.

[0027] Extraction times can range from, for example, 30 minutes to 10 days. The upper or lower limits of the extraction time may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or 1, 2, 3, 4, 5, 6, 7, 8, or 9 days. The extraction time can be adjusted as appropriate depending on the characteristics and condition of the plant being used in the extraction process.

[0028] Examples of extraction temperatures include approximately 4°C to 120°C. The upper or lower limits of the extraction temperature may be 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, or 115°C. The extraction temperature may be adjusted as appropriate depending on the characteristics and condition of the plant being used in the extraction process.

[0029] More specifically, an extraction method involves immersing the plant in an extraction solvent, allowing it to stand or be stirred as needed for 30 minutes to 10 days to dissolve the soluble components into the solvent, and then filtering to remove solid matter. Heating may also be applied as needed during the process.

[0030] When purifying the extract, methods such as activated carbon, porous resin, ion exchange resin, adsorbent, and high-performance liquid chromatography (HPLC) may be used. Alternatively, two or more purification methods may be combined.

[0031] Furthermore, as will be described later, commercially available products can also be used as the plant extract of the present invention.

[0032] Content of the plant extract of the present invention In the composition of the present invention, the content of the plant extract of the present invention is not particularly limited, but may be, for example, 0.00001 to 1% by mass, preferably 0.00005 to 0.5% by mass, more preferably 0.0001 to 0.2% by mass, even more preferably 0.0001 to 0.1% by mass, and particularly preferably 0.0001 to 0.05% by mass. The upper or lower limits of the range are, for example, 0.00005, 0.0001, 0.00015, 0.0002, 0.00025, 0.0003, 0.00035, 0.0004, 0.00045, 0.0005, 0.00055, 0.0006, 0.00065, 0.0007, 0.00075, 0.0008, 0.00085, 0.0009, 0.00095, 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045, 0 It may be 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1% by mass.

[0033] When using a commercially available plant extract as the plant extract in this invention, the commercially available product may contain solvents such as water, preservatives, etc., in addition to the plant extract. In this case, the plant extract content in the composition of this invention is calculated from the plant extract content in the commercially available product disclosed by the manufacturer of the commercially available product and the content of the commercially available product in the composition of this invention. For example, if the plant extract content in the commercially available product is 10% and the content of the said commercially available product in the composition of this invention is also 10%, then the plant extract content in the composition of this invention is calculated to be 1%.

[0034] If the composition of the present invention contains two or more plant extracts of the present disclosure, "content of plant extracts of the present invention" shall refer to the total content of each extract.

[0035] The plant extracts of the present invention will be described below.

[0036] Rose extract Roses (Rosaceae rosa) are a general term for plants belonging to the genus Rosa in the family Rosaceae. Examples of plants in the genus Rosa include Rosa centifolia, Rosa canina, Rosa damascena, Rosa gallica, Rosa multiflora, and Rosa rugosa. Rosa centifolia, Rosa canina, Rosa damascena, Rosa gallica, Rosa multiflora, and Rosa rugosa can all be preferably used as raw materials for the rose extract of the present invention. The rose extract may be used alone or in combination of two or more types. There are no particular restrictions on the part of the rose used for extraction, but it is preferable to include the flower part. The composition of the present invention preferably contains a rose extract made from Rosa centifolia among the above-mentioned roses.

[0037] In the present invention, the rose extract may be obtained by extracting from roses by the method described above, or a commercially available product containing rose extract may be used. The rose extract is "Falcorex It is contained in commercially available products such as "Rose P (manufactured by Ichimaru Falcos Co., Ltd.)", "Rose Extract S (manufactured by Koei Kogyo Co., Ltd.)", "Rose Extract BG (manufactured by Maruzen Pharmaceutical Co., Ltd.)", "Falcorex Wild Rose B (manufactured by Ichimaru Falcos Co., Ltd.)", and "Wild Rose Extract (manufactured by Koei Kogyo Co., Ltd.)". When the composition of the present invention contains rose extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "amount of plant extract of the present invention" described above, but it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.02% by mass, and even more preferably 0.001 to 0.01% by mass.

[0038] Perilla extract Perilla frutescens var. crispa is an annual herbaceous plant belonging to the genus Perilla in the family Lamiaceae. In this invention, the perilla extract may be obtained by extracting from perilla using the method described above, or a commercially available product containing perilla extract may be used. The part of the perilla used for extraction is not particularly limited, but it is preferable to include the leaves. Perilla extract is contained in commercially available products such as "Falcorex Perilla HB (manufactured by Ichimaru Falcos Co., Ltd.)", "Perilla Extract (manufactured by Koei Kogyo Co., Ltd.)", "Perilla Extract BG (manufactured by Maruzen Pharmaceutical Co., Ltd.)", "Perilla Extract-BG (manufactured by Yamada Yakken Co., Ltd.)", and "Perilla Seed Extract-LOGC (manufactured by Oryza Oil & Fat Chemical Co., Ltd.)".

[0039] If the composition of the present invention contains perilla extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "amount of plant extract of the present invention" described above, but it is preferably 0.0001 to 0.01% by mass, more preferably 0.0005 to 0.005% by mass, and even more preferably 0.0008 to 0.002% by mass.

[0040] Cherry blossom extract Cherry blossoms (Sakura) are deciduous trees or shrubs belonging to the genus Prunus in the subfamily Prunoideae of the family Rosaceae. There are many varieties of cherry blossoms, including Somei Yoshino, Yama-zakura, Oshima-zakura, Oyama-zakura, Edo-higashi, Mame-zakura, Miyama-zakura, Kasumi-zakura, Kohigan, Sato-zakura, and Kan-zakura. In this invention, the variety of cherry blossom is not particularly limited, but Somei Yoshino is particularly preferred. In this invention, the cherry blossom extract may be obtained by extracting from cherry blossoms by the method described above, or a commercially available product containing cherry blossom extract may be used. The part of the cherry blossom from which the extract is taken is not particularly limited, but it is preferable to include the leaves. Cherry blossom extract is contained in commercially available products such as "Sakura Extract B (Ichimaru Falcos Co., Ltd.)".

[0041] When the composition of the present invention contains cherry blossom extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "Content of plant extract of the present invention" described above, but it is preferably 0.0001 to 0.1% by mass, more preferably 0.001 to 0.05% by mass, and even more preferably 0.002 to 0.005% by mass.

[0042] Coffee tree extract The term "coffee tree" refers to a group of plants belonging to the genus Coffea in the family Rubiaceae. Cultivated varieties of coffee trees include Coffee arabica and Coffee robusta. While the variety of coffee tree is not particularly limited in this invention, Coffee arabica is particularly preferred. In this invention, the coffee tree extract may be obtained by extracting from the coffee tree using the method described above, or a commercially available product containing coffee tree extract may be used. While the part of the coffee tree used for extraction is not particularly limited, it is preferable to include the seeds. Coffee tree extract is found in commercially available products such as "Café Noage (Ichimaru Falcos Co., Ltd.)" and "Coffee Extract-BG (Yamada Yakken Co., Ltd.)".

[0043] If the composition of the present invention contains coffee tree extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "Content of plant extract of the present invention" described above, but it is preferably 0.0001 to 0.005% by mass, more preferably 0.0003 to 0.001% by mass, and even more preferably 0.0005 to 0.0008% by mass.

[0044] Wild Thyme Extract Wild thyme (Thymus serpyllum) is an evergreen shrub belonging to the genus Thymus in the family Lamiaceae. In this invention, the wild thyme extract may be obtained by extracting from wild thyme using the method described above, or a commercially available product containing wild thyme extract may be used. Wild thyme extract is contained in commercially available products such as "Cinderella Care (Ichimaru Falcos Co., Ltd.)", "Thyme Extract (Koei Kogyo Co., Ltd.)", and "Thyme Extract (Maruzen Pharmaceutical Co., Ltd.)".

[0045] If the composition of the present invention contains wild thyme extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.1% by mass, more preferably 0.0003 to 0.05% by mass, and even more preferably 0.0005 to 0.005% by mass.

[0046] Mayonnaise Extract Origanum majorana, also known as marjoram, is a perennial herb belonging to the genus Origanum in the Lamiaceae family. In this invention, the origanum extract may be obtained by extracting from origanum using the method described above, or a commercially available product containing origanum extract may be used. The part of the origanum used for extraction is not particularly limited, but it is preferable to include the leaves. Origanum extract is contained in commercially available products such as "Marjoram Extract BG (Maruzen Pharmaceutical Co., Ltd.)" and "Dermagenist (BASF)."

[0047] If the composition of the present invention contains marjoram extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.02% by mass, and even more preferably 0.001 to 0.01% by mass.

[0048] Chamomile extract Chamomile (Matricaria recutita, or Chamomilla recutita), also known as chamomile, is an annual plant belonging to the genus Matricaria in the family Asteraceae. In this invention, the chamomile extract may be obtained by extracting from chamomile using the method described above, or a commercially available product containing chamomile extract may be used. The part of the chamomile used for extraction is not particularly limited, but it is preferable to include the flower and / or leaf parts. Chamomile extract is contained in commercially available products such as "Chamomile Liquid (Ichimaru Falcos Co., Ltd.)", "Chamomile Extract (Koei Kogyo Co., Ltd.)", and "Chamomile Extract BG-J (Maruzen Pharmaceutical Co., Ltd.)".

[0049] When the composition of the present invention contains chamomile extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.01% by mass, and even more preferably 0.001 to 0.005% by mass.

[0050] Loquat extract The loquat (Rhaphiolepis bibas, or Eriobotrya japonica) is an evergreen tree belonging to the Rosaceae family. In this invention, the loquat extract may be obtained by extracting from the loquat by the method described above, or a commercially available product containing loquat extract may be used. The part of the loquat used for extraction is not particularly limited, but it is preferable to include the leaves. Loquat extract is contained in commercially available products such as "Falcorex Biwa Leaf B (Ichimaru Falcos Co., Ltd.)", "Loquat Extract (Koei Kogyo Co., Ltd.)", "Loquat Extract BG-J (Maruzen Pharmaceutical Co., Ltd.)", and "Loquat Leaf Extract-ET (Yamada Yakken Co., Ltd.)".

[0051] If the composition of the present invention contains loquat extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts in the Disclosure" described above, but is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.01% by mass, and even more preferably 0.001 to 0.005% by mass.

[0052] Eucalyptus extract Eucalyptus globulus is an evergreen tree belonging to the genus Eucalyptus in the family Myrtaceae. In this invention, the eucalyptus extract may be obtained by extracting from eucalyptus using the method described above, or a commercially available product containing eucalyptus extract may be used. The part of the eucalyptus from which the extract is taken is not particularly limited, but it is preferable to include the leaves. Eucalyptus extract is contained in commercially available products such as "Falcorex Eucalyptus B (Ichimaru Falcos Co., Ltd.)", "Eucalyptus Extract (Koei Kogyo Co., Ltd.)", "Eucalyptus Extract (Maruzen Pharmaceutical Co., Ltd.)", and "Eucalyptus Extract ET (Nippon Oil Co., Ltd.)".

[0053] If the composition of the present invention contains eucalyptus extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.2% by mass, more preferably 0.0005 to 0.04% by mass, and even more preferably 0.001 to 0.02% by mass.

[0054] Thyme extract Thymus vulgaris is a perennial plant belonging to the genus Thymus in the family Lamiaceae. In the present invention, the thyme extract may be obtained by extracting from thyme vulgaris by the method described above, or a commercially available product containing thyme extract may be used. The part of the thyme from which the extract is taken is not particularly limited, but it is preferable to include at least one selected from the group consisting of the flower, leaf, and stem parts. Thyme extract is contained in commercially available products such as "Thyme Extract (Koei Kogyo Co., Ltd.)", "Thyme Extract BG (Maruzen Pharmaceutical Co., Ltd.)", and "Thyme Green Extract BG (Nagahirodo Honten Co., Ltd.)".

[0055] When the composition of the present invention contains thyme extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "Content of plant extract of the present invention" described above, but it is preferably 0.0001 to 0.01% by mass, more preferably 0.0003 to 0.005% by mass, and even more preferably 0.0005 to 0.002% by mass.

[0056] Sage extract Sage (Salvia officinalis), also known as common sage, is a perennial herb or evergreen shrub belonging to the genus Salvia in the Lamiaceae family. In this invention, the sage extract may be obtained by extracting from sage using the method described above, or a commercially available product containing sage extract may be used. The part of the sage used for extraction is not particularly limited, but it is preferable to include the leaves. Sage extract is contained in commercially available products such as "Falcorex Sage B (Ichimaru Falcos Co., Ltd.)", "Salvia Extract (Koei Kogyo Co., Ltd.)", "Salvia Extract BG-J (Maruzen Pharmaceutical Co., Ltd.)", and "Sage Extract-BG (Yamada Yakken Co., Ltd.)".

[0057] If the composition of the present invention contains sage extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.01% by mass, and even more preferably 0.001 to 0.003% by mass.

[0058] Perilla extract Perilla frutescens var. crispa is an annual herbaceous plant belonging to the genus Perilla in the family Lamiaceae. In this invention, the perilla extract may be obtained by extracting from perilla using the method described above, or a commercially available product containing perilla extract may be used. The part of the perilla used for extraction is not particularly limited, but it is preferable to include the leaves and / or seeds. Perilla extract is contained in commercially available products such as "Falcorex Perilla HB (Ichimaru Falcos Co.)", "Perilla Extract (Koei Kogyo Co.)", "Perilla Extract BG (Maruzen Pharmaceutical Co.)", "Perilla Extract-BG (Yamada Yakken Co.)", and "Perilla Seed Extract-LOGC (Oryza Oil & Fat Chemical Co.)".

[0059] If the composition of the present invention contains perilla extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.5% by mass, more preferably 0.0005 to 0.1% by mass, and even more preferably 0.0008 to 0.05% by mass.

[0060] Peppermint extract Peppermint (Mentha piperita), also known as European mint, is a perennial herb belonging to the genus Mentha in the Lamiaceae family. In this invention, the peppermint extract may be obtained by extracting peppermint using the method described above, or a commercially available product containing peppermint extract may be used. The part of the peppermint used for extraction is not particularly limited, but it is preferable to include the leaves. Peppermint extract is contained in commercially available products such as "Falcorex Peppermint B (Ichimaru Falcos Co., Ltd.)", "Spring Mint (Ichimaru Falcos Co., Ltd.)", "Western Mint Extract (Koei Kogyo Co., Ltd.)", and "Peppermint Extract BG (Maruzen Pharmaceutical Co., Ltd.)".

[0061] When the composition of the present invention contains peppermint extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "Content of plant extracts of the present invention" described above, but it is preferably 0.0001 to 0.1% by mass, more preferably 0.001 to 0.02% by mass, and even more preferably 0.002 to 0.007% by mass.

[0062] Rosa Canina Extract Rosa canina, also known as dog rose, is a plant belonging to the genus Rosa in the family Rosaceae. The fruit of Rosa canina is sometimes specifically called rosehip. In this invention, the Rosa canina extract may be obtained by extracting from Rosa canina by the method described above, or a commercially available product containing Rosa canina extract may be used. There are no particular restrictions on the part of the Rosa canina from which the extract is taken, but it is preferable to include the fruit. Rosa canina extract is contained in commercially available products such as "Falcorex Nobara B (Ichimaru Falcos Co., Ltd.)", "Nobara Extract (Koei Kogyo Co., Ltd.)", and "Rosehip Extract BG100 (Maruzen Pharmaceutical Co., Ltd.)".

[0063] If the composition of the present invention contains rosehip extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.01% by mass, and even more preferably 0.001 to 0.005% by mass.

[0064] Tormentilla extract Potentilla tormentilla vulgaris, also known as Potentilla vulgaris, is a perennial herb belonging to the genus Potentilla in the family Rosaceae. In this invention, the tormentilla extract may be obtained by extracting from tormentilla using the method described above, or a commercially available product containing tormentilla extract may be used. There are no particular restrictions on the part of the tormentilla used for extraction, but it is preferable to include the roots. Tormentilla extract is contained in commercially available products such as "Falcorex Tormentilla B (Ichimaru Falcos Co., Ltd.)" and "Tormentilla Extract (Maruzen Pharmaceutical Co., Ltd.)".

[0065] If the composition of the present invention contains tormentilla extract, its content is not particularly limited and can be adjusted as appropriate by referring to the "Content of Plant Extracts of the Present Invention" described above, but it is preferably 0.00005 to 0.01% by mass, more preferably 0.0001 to 0.005% by mass, and even more preferably 0.0005 to 0.002% by mass.

[0066] Lotus extract Astragalus sinicus is a biennial plant belonging to the genus Astragalus in the family Fabaceae. In this invention, the Astragalus sinicus extract may be obtained by extracting it from Astragalus sinicus using the method described above, or a commercially available product containing Astragalus sinicus extract may be used. Astragalus sinicus extract is contained in commercially available products such as "Falcorex Astragalus sinicus (Ichimaru Falcos Co., Ltd.)".

[0067] When the composition of the present invention contains milk vetch extract, the amount is not particularly limited and can be adjusted as appropriate by referring to the "Content of plant extract of the present invention" described above, but it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.01% by mass, and even more preferably 0.001 to 0.005% by mass.

[0068] The ophthalmic product of the present invention may contain other components besides those described above, as long as they do not impair the effects of the invention. Examples of other components include, but are not limited to, surfactants, pH adjusters, cooling agents, active ingredients, stabilizers, humectants, and plant extracts other than those disclosed herein. The ophthalmic composition of the present invention may contain only one of the above-mentioned other components alone, or it may contain two or more in combination.

[0069] Examples of surfactants include anionic surfactants, amphoteric surfactants, and cationic surfactants. These surfactants can be used individually or in combination of two or more types.

[0070] Specific examples of anionic surfactants include sulfate ester salts such as sodium lauryl sulfate and sodium polyoxyethylene lauryl ether sulfate; sulfosuccinates such as sodium lauryl sulfosuccinate and sodium polyoxyethylene lauryl ether sulfosuccinate; acyl amino acid salts such as sodium lauroyl methylalanine; and sodium cocoyl methyl taurate.

[0071] Specific examples of amphoteric surfactants include, for example, betaine-type surfactants such as lauryldimethylaminoacetic acid betaine and coconut oil fatty acid amidopropyldimethylaminoacetic acid betaine; imidazoline-type surfactants such as N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine sodium; and amino acid-type surfactants such as N-lauryldiaminoethylglycine.

[0072] Specific examples of cationic surfactants include distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, benzalkonium chloride, and N-coconut oil fatty acid acyl-L-arginine ethyl DL-pyrrolidone carboxylate.

[0073] Examples of pH adjusting agents include citric acid, lactic acid, acetic acid, hydrochloric acid, potassium hydroxide, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium lactate, sodium citrate, sodium lactate, phosphoric acid, lactic acid, tartaric acid, glycerophosphate, monoethanolamine, diethanolamine, or chemically possible salts thereof. These can be used alone or in combination and can be used not only for adjusting the pH value but also for buffering the adjusted pH value.

[0074] Examples of cooling agents include, but are not limited to, menthol, camphor, limonene, and peppermint oil.

[0075] Examples of stabilizers include, but are not limited to, tocopherol, cyclodextrin, dextran, monoethanolamine, and glyceryl monostearate.

[0076] Examples of humectants include, but are not limited to, ethylene glycol, polypropylene glycol, xylitol, maltitol, lactitol, erythritol, and polyoxyethylene glycol.

[0077] The application site of the ophthalmic composition of the present invention is not particularly limited, but it is preferable to apply it to the ophthalmology and surrounding areas. Furthermore, the application form of the present invention is not particularly limited and can be used, for example, as a pharmaceutical, quasi-drug, or cosmetic, but is not limited to these.

[0078] The dosage form of the ophthalmic composition can be selected as appropriate, and examples include, but are not limited to, eye drops, contact lens insertion solutions, eye washes, artificial tears, and contact lens eye washes.

[0079] The ophthalmic composition of the present invention can be prepared by known methods or by methods readily conceivable by those skilled in the art from known methods. For example, it can be prepared by mixing the plant extract and other components of the present invention. Mixing can be carried out using a known stirrer (e.g., a homomixer). [Examples]

[0080] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these.

[0081] Test Example 1. Preparation of senescent cells 1-1. Cellular senescence was induced using normal human neonatal epidermal keratinocytes (NHEK, Lifeline Cell Technology) by known methods. Specifically, cellular senescence was induced using the following methods. Unless otherwise specified, cell culture was performed in a 37°C CO2 5v / v% incubator. (1) NHEK cells were seeded at 5.0 × 10³ cells / well in a 96-well plate and cultured for 1 day. HuMedia™-KG2 (Cat. No. KK-2150S, Kurabo Industries Ltd.) was used as the culture medium. (2) After removing the culture medium, the medium was replaced with HuMedia-KB2 (Cat. No. KK-2350S, Kurabo Industries Ltd.) containing 1.5 μM Palbociclib (Cat. No. PZ0383, Merck) and 1 v / v% dimethyl sulfoxide (DMSO), and incubated for 24 hours. (3) Remove the culture medium and replace it with HuMedia-KB2 containing 1.5 μM Palbociclib, 9 μM Nutlin-3a (Cat. No. SML0580, Merck), and 1 v / v% DMSO, and incubate for 8 hours. (4) Remove the culture medium and replace it with HuMedia-KB2 containing Nutlin-3a 9 μM, BI-2536 (Cat. No. HY-50698, MedChemExpress) 500 nM, and DMSO 1 v / v%, and incubate for 1.5 days. (5) Remove the culture medium and replace it with HuMedia-KB2 containing BI-2536 500nM and DMSO 1v / v%, and incubate for 3 days.

[0082] 1-2. The cells prepared in Test Example 1-1, and the NHEK samples before the procedure in Test Example 1-1 (i.e., untreated samples), were each stained with SA-β-gal using the following method. (1) The culture medium in the wells was aspirated and removed, and the wells were washed twice with PBS. (2) Add the 1× Fixing Solution included with the Cellular Senescence Assay Kit (Cat. No. CBA-230, CELL BIOLABS) and fix at room temperature for 5 minutes. (3) Remove the 1× Fixing Solution and wash three times with PBS. (4) The Cell Staining Working Solution included with the Cellular Senescence Assay Kit was added until the cells were completely submerged. (5) The reaction was carried out overnight in a 37°C incubator, shielded from light. (6) Remove the Cell Staining Working Solution, wash twice with PBS, and fill the wells with PBS. (7) Observe with an optical microscope to confirm whether or not it is stained blue.

[0083] 1-3 . The results of Test Example 1-2 showed that almost all cells prepared in Test Example 1-1 were SA-β-gal stain-positive, whereas no SA-β-gal stain-positive cells were observed in the NHEK cells before the procedure in Test Example 1-1. In other words, it was confirmed that cellular senescence is induced by the method described in Test Example 1-1, and that normal NHEK cells become senescent cells. Therefore, cells prepared by the method described in Test Example 1-1 were used as senescent cells in subsequent experiments. In this example, senescent cells prepared by the method described in Test Example 1-1 may be referred to as SHEK.

[0084] Test Example 2. Senescent Cell Removal Test 2-1. SHEK was cultured with or without each of the plant extracts shown in Table 1 added to the culture medium. The commercially available plant extracts shown in Table 1 were used. The concentrations (w / w%) shown in Table 1 are calculated from the content of the plant extracts of this disclosure in the commercial products and the concentration of the commercial products added to the culture medium. That is, the concentrations shown in Table 1 represent the concentration of the plant extracts of the present invention in the culture medium (sometimes referred to as "pure content concentration" in this disclosure), not the concentration of the commercial products in the culture medium.

[0085] [Table 1]

[0086] The specific method for culturing SHEK is shown below. (1) Remove the culture medium from the well containing SHEK obtained in Test Example 1-1 and wash twice with PBS. (2) HuMedia-KB2 with each plant extract shown in Table 1 added at the concentrations shown in Table 1, or HuMedia-KB2 without any plant extracts (control), was added to each well and incubated for 24 hours.

[0087] 2-2. The viability of SHEK cultured in Test Example 2-1 was measured using the Premix WST-1 Cell Proliferation Assay System (Cat. No. MK400, TaKaRa Bio, hereinafter sometimes referred to as "WST-1 Premix"). WST-1 is a reagent for quantifying cell viability by colorimetric measurement, and it utilizes the conversion of tetrazolium salt (WST-1) to formazan dye by mitochondrial dehydrogenase in living cells. An increase in the number of living cells leads to an increase in the activity of mitochondrial dehydrogenase in the sample, and this increase in enzyme activity leads to an increase in formazan dye production. Therefore, there is a linear correlation between the amount of formazan dye present and the number of metabolically active cells in the culture medium.

[0088] Specifically, the survival rate was measured using the following method. (1) Remove the culture medium from the wells containing the SHEK cultured in Test Example 2-1 and wash once with PBS. (2) HuMedia-KB2 containing 10 v / v% WST-1 premix was added to each well in 100 μL and incubated for 2 hours and 30 minutes. (3) For each well, the absorbance at 450 nm was measured using an xMark® Microplate Spectrophotometer (BIO RAD). (4) Based on the measured absorbance at 450 nm, the cell viability when each plant extract was added was calculated using the following formula, with the cell viability when no plant extract was added (control) set to 1. As mentioned above, there is a linear correlation between the number of metabolically active cells (i.e., viable cells) and the formazan dye that absorbs at 450 nm. Cell viability = absorbance at 450 nm with plant extract added / absorbance at 450 nm without plant extract (control). The test was performed with n=5. The results are shown in Figures 1 and 2, and Tables 2 and 3. In Figures 1 and 2, the top of the bar graph represents the mean value, and the error bars represent ± standard deviation.

[0089] 2-3. NHEK that had not undergone senescence induction was also cultured with or without the plant extracts shown in Table 1 added to the culture medium. Specifically, NHEK was cultured using the following method. (1) NHEK cells were seeded at a rate of 1.0 × 10⁴ cells / well in a 96-well plate and cultured for 1 day. HuMedia-KG2 was used as the culture medium. (2) HuMedia-KB2 with each plant extract shown in Table 1 added at the concentrations shown in Table 1, or HuMedia-KB2 without any plant extracts (control), was added to each well in 100 μL and incubated for 24 hours.

[0090] 2-4. For the NHEK cultured in Test Example 2-3, the viability was measured using the WST-1 premix, similar to Test Example 2-2. Specifically, the viability was measured using the following method. (1) Remove the culture medium from the wells containing NHEK cultured in Test Examples 2-3 and rinse once with PBS. It was cleaned. (2) HuMedia-KB2 containing 10 v / v% WST-1 premix was added to each well in 100 μL and incubated for 2 hours and 30 minutes. (3) For each well, the absorbance at 450 nm was measured using an xMark Microplate Spectrophotometer. (4) From the measured absorbance at 450 nm, the cell viability when each plant extract was added was calculated based on the following formula, with the cell viability when no plant extract was added (control) set to 1. Cell viability = absorbance at 450 nm with plant extract added / absorbance at 450 nm without plant extract (control). The test was performed with n=5. The results are shown in Figures 1 and 2, and Tables 2 and 3. In Figures 1 and 2, the top of the bar graph represents the mean value, and the error bars represent ± standard deviation.

[0091] Whether a component has a senescent cell-removing effect was determined if the survival rate of senescent cells cultured with the component added was lower than the survival rate of senescent cells cultured without the component.

[0092] [Table 2]

[0093] [Table 3]

[0094] 2-5. Discussion As shown in Figures 1 and 2, and Tables 2 and 3, when each plant extract was added to the SHEK culture medium, the cell viability was lower compared to when the plant extract was not added. This result suggests that each plant extract has an effect of removing senescent cells.

[0095] Furthermore, when each plant extract was added, SHEK resulted in a lower cell viability rate than NHEK. This result suggests that each plant extract can selectively induce cell death in senescent cells.

Claims

1. An ophthalmic composition containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and lotus.

2. An anti-aging ophthalmic composition containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus.

3. An ophthalmic composition for improving dry eye, containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and astragalus.

4. An ophthalmic composition for improving blurred vision, containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and lotus.

5. An ophthalmic composition for improving eye strain, containing at least one plant extract selected from the group consisting of wild thyme, rose, marjoram, chamomile, loquat, eucalyptus, thyme, sage, perilla, cherry, coffee tree, peppermint, rosehip, tormentilla, and lotus.

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