WO2013031003A1 - Dna repair accelerator and dna damage suppressant - Google Patents

Abstract

A DNA repair accelerator containing a cinchona extract, and a DNA damage suppressant containing at least one from among a cinchona extract, a comfrey extract, an Coffea extract, a Pueraria root extract, a burdock extract, a Coptis extract, a Sophora angustifolia extract, a chlorella extract, a lavender extract, an Oenothera biennis extract, a rose extract, a Gynostemma pentaphyllum extract, a Rabdosia japonica extract, a Lamium album extract, a carrot extract, a Tilia japonica extract, a great burnet extract, a lotus extract, a tea extract and an Abelmoschus esculentus extract.

Description

DNA repair promoter and DNA damage inhibitor

The present invention relates to a DNA repair promoter and a DNA damage inhibitor.

When the skin is exposed to ultraviolet rays, DNA damage may be caused in the skin cells. At this time, damaged DNA such as cyclobutane-type pyrimidine dimer and 6-4 photoproduct is formed in the DNA of skin cells. Such damaged DNA is usually repaired by a living body DNA repair mechanism. However, when DNA repair is not normally performed in vivo, skin injury, skin aging, cell apoptosis, cell canceration, etc. may be caused.

Therefore, cosmetics for protecting the skin from ultraviolet rays such as sunscreen agents that block ultraviolet rays have been developed in order to suppress the adverse effects of the ultraviolet rays on the skin. However, in daily life, the sunscreen agent applied to the skin is peeled off due to sweating or the like, so that a sufficient effect may not be obtained.

On the other hand, in order to promote the repair of damaged DNA in the skin, a DNA repair promoter containing an extract of Gigartina tenella has been proposed (for example, see Patent Document 1). In order to prevent DNA damage, Sarcandra glabra, Saraca dives, Cudrania pubescens, Taxodidium distium, Rubydiac, and Rubydiac. tonkinensis, Alcornea trebioides, Berchemia polyphylla, Grochidion publicum, Sassafrassum Tzumu) DNA damaging inhibitor comprising an extract of a plant, such as has been proposed (e.g., see Patent Document 2).

However, there is a need for other DNA repair promoters and DNA damage inhibitors that are excellent in promoting repair of damaged DNA and / or suppressing the production of damaged DNA.

JP 2006-273761 A JP 2008-247854 A

The present invention has been made in view of the above prior art, and an object of the present invention is to provide a DNA repair promoter excellent in promoting the repair of damaged DNA. Another object of the present invention is to provide a DNA damage inhibitor excellent in inhibitory properties against the generation of damaged DNA.

That is, the present invention
(1) a DNA repair promoter that promotes repair of damaged DNA, comprising a kina extract, and (2) a DNA damage inhibitor that suppresses the generation of damaged DNA. Kina extract, Comfrey extract, Coffea extract, Cuckoo extract, Burdock extract, Auren extract, Clara extract, Chlorella extract, Lavender extract, Evening primrose extract, Rose extract, Achachan extract Containing at least one extract selected from the group consisting of edible extract, enamel extract, carrot extract, carrot extract, linden extract, gill extract, lotus extract, tea extract and okra extract To a DNA damage inhibitor characterized by

The DNA repair promoter of the present invention has the effect of being excellent in promoting repair of damaged DNA. In addition, the DNA damage inhibitor of the present invention has an effect of being excellent in suppressing the generation of damaged DNA.

6 is a graph showing the relationship between the elapsed time from the end of UV-B irradiation and the CPD residual rate in Test Example 2.

1. DNA repair promoter The DNA repair promoter of the present invention is a DNA repair promoter that promotes the repair of damaged DNA, and is characterized by containing a kina extract.

In the present specification, “damaged DNA” refers to a polynucleotide containing a modified nucleoside and a modified nucleoside residue generated by DNA damage. Examples of the modified nucleoside include cyclobutane type pyrimidine dimer (CPD), 6-4 photoproduct (6-4PP) [eg, pyrimidine (6-4) pyrimidone dimer, etc.] and the like. DNA damage occurs, for example, when normal DNA of cells is exposed to ultraviolet rays, active oxygen, mutagenic substances, and the like.

Since the DNA repair promoter of the present invention contains a kina extract, it exhibits excellent facilitation for repairing damaged DNA.

Further, since the kina extract is an extract obtained from a kina plant that is not substantially burdened on humans, it can be used as a raw material for foods, crude drugs and the like. Therefore, the DNA repair promoter of the present invention containing the extract has an excellent property that the load on humans is small.

The kina extract is a component that promotes the repair of damaged DNA or a component that suppresses the generation of damaged DNA.

Examples of the part of the kina plant that is the plant material of the kina extract include bark. In the extraction, the plant raw material may be used as it is, or may be used after being dried. Moreover, you may grind | pulverize and use a plant raw material. The extraction can be performed, for example, by immersing the plant material in an extraction solvent and heating and / or pressurizing as necessary. Further, the extraction temperature and the extraction time cannot be determined unconditionally because they vary depending on the type of extraction solvent, the type of plant, the use of the extract, and the like.

As an extraction solvent, water; lower alcohols such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin; diethyl ether, dipropyl ether Ethers such as: acetates such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone. These extraction solvents may be used alone or in combination of two or more.

The kina extract can be obtained, for example, by extracting bark of a genus Kina [for example, Cinchona succirubra] using an extraction solvent such as water or ethyl alcohol. The quina extract contains, for example, quinidine, quinine, cinchonine and the like. In the present invention, a commercially available kina extract may be used as the kina extract.

The DNA repair promoter of the present invention may be in a dosage form that does not contain substances other than the kina extract, such as a dried kina extract, and the dried state in a solvent such as purified water or a buffer solution. It may be a liquid dosage form in which the kina extract is mixed.

When the DNA repair accelerator of the present invention is the liquid agent, the content of the dried kina extract in the DNA repair accelerator varies depending on the use of the DNA repair accelerator, so it can be determined in general. Since it cannot be performed, it is preferable to set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried kina extract in the DNA repair promoter of the present invention is preferably 0.00003% by mass or more from the viewpoint of sufficiently securing the repair of damaged DNA. From the viewpoint of ease of handling during use, it is preferably 3% by mass or less, and more preferably 0.3% by mass or less.

The DNA repair promoter of the present invention may contain, for example, an auxiliary agent for expressing the acceleration of repair of damaged DNA by a kina extract, if necessary.

As used herein, the facilitation of repair of damaged DNA refers to, for example, damage relative to the rate of decrease of damaged DNA in cells not contacted with the DNA repair promoter of the present invention (eg, cells contacted with a control). From the viewpoint of ensuring higher acceleration of DNA repair, assuming that the reduction rate of damaged DNA in cells in contact with the control is 100, the reduction rate of damaged DNA is 110 or more, preferably 150 or more, more preferably Means to promote repair of damaged DNA so as to be 200 or more.

The rate of decrease of the damaged DNA is determined by, for example, contacting the DNA repair promoter or control of the present invention with a cell group, quantifying the amount of damaged DNA in the obtained treated cells over time, and measuring damaged DNA per unit time. It can be obtained by calculating the amount of change in the amount.

The amount of damaged DNA is, for example, (A) a step of fixing a cell group to obtain a fixed cell group,
(B) a step of subjecting the cells of the fixed cell group obtained in the step (A) to DNA denaturation treatment to obtain a treated cell group;
(C) the treated cell group obtained in the step (B), an anti-damaged DNA antibody, an antibody detection reagent that specifically binds to the anti-damaged DNA antibody and emits fluorescence, and the cells of the cell group Combining a cell staining reagent that binds and emits fluorescence to cause the damaged DNA in the cell to react with the anti-damaged DNA antibody and bind the cell and the cell staining reagent;
(D) Measure the intensity of fluorescence emitted from the antibody detection reagent bound to the anti-damaged DNA antibody in the mixture obtained in step (C) and the intensity of fluorescence emitted from the cell staining reagent bound to cells. And (E) normalizing the intensity of the fluorescence emitted from the antibody detection reagent measured in the step (D) by the intensity of the fluorescence emitted from the cell staining reagent measured in the step (D), It can be determined by a quantification method including a step of calculating the amount of damaged DNA from the normalized fluorescence intensity.

Since the amount of the DNA repair promoter of the present invention used at the time of contact with an object that promotes repair of damaged DNA differs depending on the use of the DNA repair promoter of the present invention, it cannot be determined unconditionally. It is preferable to set appropriately according to the use of the DNA repair inhibitor of the present invention.

Since the DNA repair promoter of the present invention can promote repair of damaged DNA, it is useful for, for example, the use of reducing or reducing damaged DNA in human tissues to improve or prevent tissue conditions caused by damaged DNA. It is.

2. DNA damage inhibitor The DNA damage inhibitor of the present invention is a DNA damage inhibitor that suppresses the generation of damaged DNA, and is a kina extract, a comfrey extract, a coffee tree extract, a cuckoo extract, a burdock extract, an auren Extract, Clara extract, Chlorella extract, Lavender extract, Evening primrose extract, Rose extract, Achazul extract, Enmezo extract, Oyster extract, Carrot extract, Linden extract, Jade extract, Lotus extract And at least one extract selected from the group consisting of a green tea extract and an okra extract.

Since the DNA damage inhibitor of the present invention contains the extract, it exhibits excellent inhibitory properties against the generation of damaged DNA.

Further, since the extract is an extract obtained from a plant that is not substantially burdened on humans, it can be used as a raw material for foods, crude drugs and the like. Therefore, the DNA repair promoter of the present invention containing the extract has an excellent property that the load on humans is small.

The extract includes kina (Kina genus plant), comfrey, coffee tree (Coffea genus plant), cuckoo (waste), burdock, auren, clara, chlorella, lavender, evening primrose, rose, amacha eel, enmiso, odorifera, carrot (garden ginseng) ), Linden (bodaiju), jiyu (waremoko), lotus, tea and okra.

Examples of plant parts that are plant raw materials for the extract include leaves, flowers, seeds, roots, stems, and buds. In the extraction, the plant raw material may be used as it is, or may be used after being dried. Moreover, you may grind | pulverize and use a plant raw material. The extraction can be performed, for example, by immersing the plant material in an extraction solvent and heating and / or pressurizing as necessary. In addition, the extraction temperature and extraction time vary depending on the type of extraction solvent, the type of plant, the use of the extract, etc., and cannot be determined unconditionally, so the type of extraction solvent, the type of plant, the use of the extract, etc. It is preferable to set appropriately according to the above.

As an extraction solvent, water; lower alcohols such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin; diethyl ether, dipropyl ether And ethers; esters such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone. These extraction solvents may be used alone or in combination of two or more.

The kina extract is the same as the kina extract used for the DNA repair promoter.

Comfy extract is a component that suppresses the generation of damaged DNA. The Comfrey extract is an extract obtained by extracting Comfrey leaves, roots, and the like using an extraction solvent such as water, ethyl alcohol, and isobutyl alcohol. The Comfrey extract contains, for example, Alainton, komsoridine, rosmarinic acid, pantothenic acid, tannin, nicotinic acid and the like. In the present invention, a commercially available Comfrey extract may be used as the Comfrey extract.

Coffee extract is a component that suppresses the generation of damaged DNA. A coffee tree extract is an extract obtained by extracting seeds of a plant of the genus Coffea using an extraction solvent such as an acidic aqueous solution of ascorbic acid or an acidic aqueous solution of citric acid. The coffee tree extract contains, for example, chlorogenic acid, polyphenol and the like. In the present invention, a commercially available coffee tree extract may be used as the coffee tree extract.

Cuckoo extract is a component that suppresses the generation of damaged DNA. The kakonkon extract is an extract obtained by, for example, extracting waste roots using an extraction solvent such as 1,3-butylene glycol and water. The kacon extract contains, for example, isoflavone glycosides and saponins. In the present invention, a commercially available cool extract may be used as the cool extract.

Burdock extract is a component that suppresses the generation of damaged DNA. The burdock extract is, for example, an extract obtained by extracting burdock root and the like using an extraction solvent such as ethyl alcohol and water. Burdock extract contains, for example, inulin, tannin, polysaccharides and the like. In the present invention, a commercially available burdock extract may be used as the burdock extract.

ウ Ouren extract is a component that suppresses the generation of damaged DNA. The auren extract is, for example, an extract obtained by extracting the auricular rhizome and the like using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The auren extract contains, for example, berberine, aurenin, palmatin and the like. In the present invention, a commercially available auren extract may be used as the auren extract.

Clara extract is a component that suppresses the generation of damaged DNA. The Clara extract is, for example, an extract obtained by extracting the roots of Clara using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The Clara extract contains, for example, matrine, oxymatrine, triphorylhidine and the like. In the present invention, a commercially available Clara extract may be used as the Clara extract.

Chlorella extract is a component that suppresses the generation of damaged DNA. The chlorella extract is, for example, an extract obtained by extracting a chlorella alga body using an extraction solvent such as 1,3-butylene glycol and water. The chlorella extract contains, for example, glycoproteins, nucleic acid-related substances, polysaccharides and the like. In the present invention, a commercially available chlorella extract may be used as the chlorella extract.

Lavender extract is a component that suppresses the generation of damaged DNA. The lavender extract is, for example, an extract obtained by extracting lavender flowers and the like using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The lavender extract contains, for example, tannin, linalool, limonene and the like. In the present invention, a commercially available lavender extract may be used as the lavender extract.

The evening primrose extract is a component that suppresses the generation of damaged DNA. The evening primrose extract is, for example, an extract obtained by extracting seeds of evening primrose using an extraction solvent such as 1,3-butylene glycol. The evening primrose extract contains, for example, polyphenols. In the present invention, a commercially available evening primrose extract may be used as the evening primrose extract.

Rose extract is a component that suppresses the generation of damaged DNA. The rose extract is, for example, an extract obtained by extracting petals of rose genus plants using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The rose extract contains, for example, eugenin. In the present invention, a commercially available rose extract may be used as the rose extract.

Amateur extract is a component that suppresses the generation of damaged DNA. The amateur extract is, for example, an extract obtained by extracting leaves, whole plants, etc. of the genus Achacha using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The amateur extract contains, for example, saponin. In the present invention, a commercially available amateur extract may be used as the amateur extract.

An enamel extract is a component that suppresses the generation of damaged DNA. An enamel extract is, for example, an extract obtained by extracting leaves of an enamel tree using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. An enamel extract contains, for example, enmain, isodocarpine, nodocin, orinidon and the like. In the present invention, a commercially available enamel extract may be used as the enamel extract.

Physarum extract is a component that suppresses the generation of damaged DNA. The nettle extract is, for example, an extract obtained by extracting flowers, stems, leaves and the like using a solvent such as 1,3-butylene glycol, ethyl alcohol and water. The nettle extract contains, for example, tannins, flavonoids and the like. In the present invention, a commercially available Amaranthus extract may be used as the Astragalus extract.

The carrot extract is a component that suppresses the generation of damaged DNA. The carrot extract is, for example, an extract obtained by extracting roots of ginseng using an extraction solvent such as propylene glycol and water. The carrot extract contains, for example, vitamins, malic acid, amino acids or derivatives thereof, sugars, pectin, lecithin and the like. In the present invention, a commercially available carrot extract may be used as the carrot extract.

Linden extract is a component that suppresses the generation of damaged DNA. The linden extract is, for example, an extract obtained by extracting flowers, leaves, and the like of bodaiju using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The linden extract contains, for example, tannins, flavonoids and the like. In the present invention, a commercially available linden extract may be used as the linden extract.

Jiyu extract is a component that suppresses the generation of damaged DNA. The citrus extract is, for example, an extract obtained by extracting the roots, rhizomes, and the like of an extract using an extraction solvent such as 1,3-butylene glycol, ethyl alcohol, and water. The jelly extract contains, for example, tannin, saponin and the like. In the present invention, a commercially available jelly extract may be used as the jelly extract.

The lotus extract is a component that suppresses the generation of damaged DNA. The lotus extract is an extract obtained by, for example, extracting lotus leaves, germs and the like using an extraction solvent such as 1,3-butylene glycol and water. The lotus extract contains, for example, demethylcoclaurine, luciferin and the like. In the present invention, a commercially available lotus extract may be used as the lotus extract.

Tea extract is a component that suppresses the generation of damaged DNA. For example, tea extract may be extracted from tea leaves using an extraction solvent such as water, acidic aqueous solution, ethyl alcohol, ethyl alcohol aqueous solution, methyl alcohol, methyl alcohol aqueous solution, acetone, ethyl acetate, glycerin aqueous solution. Is an extract obtained by The tea extract contains, for example, catechin, theanine, tannin and the like. In the present invention, a commercially available tea extract may be used as the tea extract.

Okra extract is a component that suppresses the generation of damaged DNA. The okra extract is an extract obtained by extracting, for example, okra fruits and seeds using an extraction solvent such as water. The okra extract contains, for example, polysaccharides. In the present invention, a commercially available okra extract may be used as the okra extract.

In addition, as long as the objective of this invention is not inhibited, you may use another solvent as said extraction solvent.

Among the extracts used for the DNA damage inhibitor of the present invention, a comfrey extract, a coffee tree extract, a cuckoo extract, a clara extract, a chlorella extract are used from the viewpoint of ensuring higher suppression of the generation of damaged DNA. , Lavender extract, evening primrose extract, rose extract, macaque extract, enamel extract, nettle extract, carrot extract, linden extract, gill extract, lotus extract, tea extract, okra extract , Comfrey extract, coffee tree extract, cuckoo extract, clara extract, chlorella extract, evening primrose extract, enmiso extract, ginger extract, lotus extract, tea extract, okra extract are more preferable. Free extract, coffee tree extract, cuckoo extract, lotus extract, okra extract Preferred.

The DNA repair promoter of the present invention may be in a dosage form that does not contain substances other than the extract, such as a dried product of the extract, and the extract of the extract in a solvent such as purified water or a buffer solution. It may be a liquid dosage form in which a dried product is mixed.

When the DNA repair accelerator of the present invention is the liquid agent, the content of the dried product of the extract in the DNA repair accelerator cannot be determined unconditionally because it varies depending on the use of the DNA repair accelerator. Therefore, it is preferable to set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the extract in the DNA damage inhibitor of the present invention is preferably 0.000001% by mass or more, and 0.000003% by mass from the viewpoint of sufficiently promoting the ability to repair damaged DNA. % Or more is more preferable, and from the viewpoint of easy handling during use, it is preferably 5% by mass or less, and more preferably 3% by mass or less.

When the DNA repair promoter of the present invention is the above liquid preparation containing a quina extract, the content of the dried quina extract in the quina extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the kina extract in the DNA damage inhibitor of the present invention is preferably 0.00003% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of ease of handling during use, it is preferably 3% by mass or less, and more preferably 0.3% by mass or less. In addition, when using a DNA damage inhibitor containing a kina extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the liquid containing the Comfrey extract, the content of the dried product of the Comfrey extract in the Comfrey extract in the DNA damage inhibitor of the present invention is the DNA repair Since it depends on the use of the accelerator and cannot be determined in general, it is preferably set as appropriate according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the comfrey extract in the DNA damage inhibitor of the present invention is preferably 0.000015% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA. The amount is more preferably 0001% by mass or more, and preferably 1.5% by mass or less, more preferably 0.15% by mass or less from the viewpoint of easy handling during use. When a DNA damage inhibitor containing a comfrey extract is used, the amount used at the time of contact with an object that suppresses the generation of damaged DNA differs depending on the use of the DNA damage inhibitor of the present invention, so it is general. Therefore, it is preferable to set appropriately depending on the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned liquid preparation containing a coffee tree extract, the content of the dried coffee tree extract in the coffee tree extract of the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the coffee tree extract in the DNA damage inhibitor of the present invention is preferably 0.000006% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 0.6% by mass or less, and more preferably 0.06% by mass or less. In addition, when using a DNA damage inhibitor containing a coffee tree extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing a cuspid extract, the content of the dried product of the cuspid extract in the cuspid extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the cold extract in the DNA damage inhibitor of the present invention is preferably 0.000003% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 0.3% by mass or less, and more preferably 0.03% by mass or less. In addition, when using a DNA damage inhibitor containing a cuckoo extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing burdock extract, the content of the dried product of burdock extract in the burdock extract of the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the burdock extract in the DNA damage inhibitor of the present invention is preferably 0.000011% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, 1.1% by mass or less is preferable, and 0.11% by mass or less is more preferable. In addition, when using a DNA damage inhibitor containing burdock extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned liquid preparation containing an auren extract, the content of the dried product of the aurene extract in the auren extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the auren extract in the DNA damage inhibitor of the present invention is preferably 0.000005% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory effect on the generation of damaged DNA, and is 0.00001. From the viewpoint of easy handling during use, 0.5% by mass or less is preferable, and 0.05% by mass or less is more preferable. In addition, when using a DNA damage inhibitor containing an aurene extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing the Clara extract, the content of the dried product of the Clara extract in the Clara extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the Clara extract in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. In addition, when using a DNA damage inhibitor containing Clara extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-described solution containing a chlorella extract, the content of the dried product of the chlorella extract in the chlorella extract of the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the chlorella extract in the DNA damage inhibitor of the present invention is preferably 0.0000035% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA, and is 0.00001. From the viewpoint of easy handling during use, it is preferably 0.35% by mass or less, and more preferably 0.035% by mass or less. When a DNA damage inhibitor containing a chlorella extract is used, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention. Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing a lavender extract, the content of the dried product of the lavender extract in the lavender extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the lavender extract in the DNA damage inhibitor of the present invention is preferably 0.0000035% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory effect on the generation of damaged DNA, and is 0.00001. From the viewpoint of easy handling during use, it is preferably 0.35% by mass or less, and more preferably 0.035% by mass or less. In addition, when using a DNA damage inhibitor containing a lavender extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing an extract of evening primrose, the content of the dried extract of the evening primrose extract in the DNA damage inhibitor of the present invention is the content of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried extract of the evening primrose extract in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. In addition, when using a DNA damage inhibitor containing an evening primrose extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing a rose extract, the content of the dried rose extract in the rose extract of the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the rose extract in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. When using a DNA damage inhibitor containing a rose extract, the amount used at the time of contact with an object that suppresses the production of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the liquid preparation containing an amateur extract, the content of the dried extract of the armature extract in the amateur damage extract of the DNA damage suppressor of the DNA repair promoter Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the armature extract in the DNA damage inhibitor of the present invention is preferably 0.000005% by mass or more, from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA, 0.00001 From the viewpoint of easy handling during use, 0.5% by mass or less is preferable, and 0.05% by mass or less is more preferable. In addition, when using a DNA damage inhibitor containing an amacha extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing an enamel extract, the content of the dried product of the enamel extract in the enamel extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the Amaranthus extract in the DNA damage inhibitor of the present invention is preferably 0.000008% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA, and is 0.00001. From the viewpoint of easy handling during use, 0.8% by mass or less is preferable, and 0.08% by mass or less is more preferable. In the case of using a DNA damage inhibitor containing an enamel extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned liquid preparation containing the extract of nettle perilla, the content of the dried product of the nettle extract in the nettle extract of the DNA damage suppressor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. In general, the content of the dried product of the extract of Oetalia in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. In addition, when using a DNA damage inhibitor containing a nettle extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention. Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing a carrot extract, the content of the dry product of the carrot extract in the carrot extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the carrot extract in the DNA damage inhibitor of the present invention is preferably 0.0000075% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA, and is 0.00001. From the viewpoint of easy handling during use, it is preferably 0.75% by mass or less, and more preferably 0.075% by mass or less. In addition, when using a DNA damage inhibitor containing a carrot extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA differs depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the liquid preparation containing a linden extract, the content of the dried linden extract in the linden extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried linden extract in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. In addition, when using a DNA damage inhibitor containing a linden extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention. Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-described liquid preparation containing a jujube extract, the content of the dried extract of the juju extract in the juju extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried extract of the juice extract in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. In the case of using a DNA damage inhibitor containing a citrus extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing a lotus extract, the content of the dried lotus extract in the lotus extract of the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried lotus extract in the DNA damage inhibitor of the present invention is preferably 0.00001% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1% by mass or less, and more preferably 0.1% by mass or less. In addition, when using a DNA damage inhibitor containing a lotus extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing a tea extract, the content of the dried tea extract in the tea extract in the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dry product of the tea extract in the DNA damage inhibitor of the present invention is preferably 0.000016% by mass or more from the viewpoint of sufficiently expressing the inhibitory property against the generation of damaged DNA. From the viewpoint of easy handling during use, it is preferably 1.6% by mass or less, and more preferably 0.16% by mass or less. In addition, when using a DNA damage inhibitor containing a tea extract, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention, so it is generally Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

When the DNA repair promoter of the present invention is the above-mentioned solution containing an okra extract, the content of the dried product of the okra extract in the okra extract of the DNA damage inhibitor of the present invention is the amount of the DNA repair promoter. Since it varies depending on the use and cannot be determined in general, it is preferably set appropriately according to the use of the DNA damage inhibitor. Usually, the content of the dried product of the okra extract in the DNA damage inhibitor of the present invention is preferably 0.0000035% by mass or more from the viewpoint of sufficiently exhibiting the inhibitory property against the generation of damaged DNA, and is 0.00001. From the viewpoint of easy handling during use, it is preferably 0.35% by mass or less, and more preferably 0.035% by mass or less. When a DNA damage inhibitor containing okra extract is used, the amount used at the time of contact with an object that suppresses the generation of damaged DNA varies depending on the use of the DNA damage inhibitor of the present invention. Since it cannot be determined, it is preferable to set appropriately according to the use of the DNA damage inhibitor of the present invention.

The DNA damage inhibitor of the present invention may contain, for example, an auxiliary agent for expressing an inhibitory property against the production of damaged DNA by the extract.

In the present specification, the inhibitory property against the production of damaged DNA is, for example, the amount (in moles) of damaged DNA in a cell not contacted with the DNA damage inhibitor of the present invention (for example, a cell contacted with a control). From the viewpoint of securing a higher suppression of the generation of damaged DNA, the amount of damaged DNA (in moles) should be at least 90, preferably 80 or less, more preferably 70 or less. It means to suppress generation.

Since the DNA damage inhibitor of the present invention can suppress the production of damaged DNA, it is useful for, for example, the use of reducing or reducing damaged DNA in human tissues to improve or prevent tissue conditions caused by damaged DNA. is there.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to such examples.

(Test Example 1)
To 100 μL of medium (Wako Pure Chemical Industries, Ltd., trade name: D-MEM) in each well of a 96-well plate (trade name: 96-well microplate (flat bottom), manufactured by Narje Nunk International Co., Ltd.) × 10 ⁴ cells of normal human fibroblasts were seeded. Thereafter, the 96-well plate seeded with fibroblasts was placed in a CO ₂ incubator kept at a carbon dioxide concentration of 5% by volume and 37 ° C. and incubated for 1 day.

Next, a medium [Wako Pure Chemical Industries, Ltd., trade name: D-MEM] 100 μL was further added to each well of the 96-well plate containing the cultured fibroblasts, and the 96-well after the addition of the medium. Plates were incubated for 1 day to be 100% confluent as before.

Thereafter, the medium was removed from each well of the 96-well plate containing the cultured cells, and the cells in each well were washed twice with 100 μL of physiological saline.

Corresponding to the amount of damaged DNA generated in the DNA when the washed cells are irradiated with 40 mJ / m ² of UV-C from an upper position 28 to 40 cm away from the surface of the medium. An amount of UV-B was irradiated.

A test sample-containing medium was added to a 96-well plate containing fibroblasts after irradiation with UV-B, and incubated in a CO ₂ incubator maintained at a carbon dioxide concentration of 5% by volume and 37 ° C. In addition, about 150 kinds of plant-derived extracts were used as test samples. The concentration of the test sample in the test sample-containing medium is 1% by mass.

Next, 100 μL of a cell fixing solution [methyl alcohol-acetone mixed solvent (methyl alcohol / acetone (volume ratio) = 1/1), −20 ° C.] is added to the well of the 96-well plate, and the 96-well plate is added to room temperature. The fibroblasts in the wells were fixed by allowing them to stand for 10 minutes.

Thereafter, each well of the 96-well plate was washed twice with a washing solution (composition: 10 mM phosphate buffered physiological saline), and further washed once with ultrapure water.

In each well of the washed 96-well plate, 100 μL of 2M hydrochloric acid aqueous solution was added, and the 96-well plate was allowed to stand at room temperature for 30 minutes for DNA denaturation treatment.

Thereafter, each well of the 96-well plate was washed three times with a washing solution (composition: 10 mM phosphate buffered physiological saline).

After adding 150 μL of blocking solution (phosphate buffered physiological saline containing 1% by weight skim milk) to each well of the 96-well plate after washing, the 96-well plate was allowed to stand at 37 ° C. for 30 minutes, Blocking was performed.

Thereafter, each well of the 96-well plate was washed three times with a washing solution (composition: 10 mM phosphate buffered physiological saline).

In each well of the 96-well plate after washing, a 200-fold dilution of a primary antibody solution containing an anti-CPD antibody as a primary antibody [primary antibody concentration (protein concentration standard): 88.5 μg / 10 mM phosphate buffered physiological saline] 1 mL] After adding 40 μL, the 96-well plate was allowed to stand at 37 ° C. for 60 minutes. As the anti-CPD antibody, a commercially available anti-CPD antibody [manufactured by Cosmo Bio Co., Ltd., trade name: Anti-Cyclobutane pyrimidine dimmers (CPDs), code number: NMDND001] is used as clone CDM TDM-2. In the literature [Toshio Mori et al., “Cyclobutane pyrimidine dimer-specific or (6-4) photoproduct-specific monoclonal antibody derived from the same mouse immunized with UV-irradiated DNA [Simultaneous Establishment of Monoclonal antigenic speci fi cial forefrontier therapeutics]. dimer or (6-4) photoproduct from he same mouse immunized with ultraviolet-irradiated DNA.] ", photo Chemistry and photo chemistry (Photochem.Photobiol.), Vol. 54, p. 225-232 and Tsukasa Matsunaga et al., “(6-4) Establishment and characterization of monoclonal antibodies that recognize the dewar isomers of photoproducts (Establishment and charac- terization of monoclonal anti-body recombination the dewarsomizers 6”). ], Photochemistry and Photochemistry, Vol. 57, p. 934-940] and an antibody (TDM-2) having the same properties as the commercially available anti-CPD antibody was used.

The 96-well plate after standing was washed 3 times with a washing solution (composition: 10 mM phosphate buffered physiological saline).

A biotin-labeled anti-mouse IgG antibody [manufactured by Invitrogen Corp., trade name: Biotin-Goat F (ab ′) ₂ anti-mouse IgG (H + L) (catalog number) 62-6340)] in a 200-fold dilution of secondary antibody solution (secondary antibody concentration (protein concentration standard): 2.0 μg / 10 mM phosphate buffered saline 1 mL) Thereafter, the 96-well plate was allowed to stand at 37 ° C. for 30 minutes.

Thereafter, each well of the 96-well plate was washed with 100 μL of a washing solution (composition: 10 mM phosphate buffered physiological saline) three times.

This labeled streptavidin (labeled streptavidin concentration: 1.0 mg / mL) is a labeled streptavidin solution that is a tertiary reagent containing labeled streptavidin (manufactured by LI-COR Biosciences, Inc., trade name: IRDye 800CW Labeled Streptavidin). The cell staining reagent (manufactured by LI-COR Biosciences, Inc., trade name: Sapphire700 Stain, 10 mM phosphoric acid so that the concentration of labeled streptavidin in the avidin solution is 1/2500 times the concentration. Dilution diluted with buffered saline) was added to obtain a mixture. Next, 50 μL of the mixture was added to each well of the washed 96-well plate, and the 96-well plate was allowed to stand at 37 ° C. for 30 minutes.

Thereafter, each well of the 96-well plate was washed with 100 μL of a washing solution (composition: 10 mM phosphate buffered physiological saline) three times.

The fluorescence intensity at wavelengths of 680 nm and 780 nm of the mixture in each well of the 96-well plate after washing was measured. The same operation as described above was performed except that control fibroblast NHF was used in place of the fibroblasts irradiated with UV-B, and the mixture in the wells at wavelengths of 680 nm and 780 nm, respectively. The fluorescence intensity was measured. In addition, after incubating the 96-well plate containing cells after UV-B irradiation, instead of adding the cell fixing solution to the wells, the 96-well plate containing cells immediately after UV-B irradiation was added. Except that the cell fixing solution was added to the well, the same operation as described above was performed, and the fluorescence intensity of the mixture in the well at wavelengths of 680 nm and 780 nm was measured. Then, a value obtained by subtracting the fluorescence intensity value at a wavelength of 780 nm when UV-B was not irradiated from the fluorescence intensity value at a wavelength of 780 nm when UV-B was irradiated as ultraviolet rays was calculated. Furthermore, the fluorescence intensity based on the labeled streptavidin labeling substance at the excitation wavelength of 780 nm in each well was normalized by the fluorescence intensity based on the cell staining reagent at the excitation wavelength of 680 nm in each well, and the error between wells was corrected. Thereafter, the amount of CPD was calculated based on a calibration curve prepared in advance.

The CPD residual rate when a predetermined time has elapsed from the end of UV-B irradiation was determined by setting the amount of CPD in fibroblast DNA immediately after UV-B irradiation to 100. From about 150 types of plant-derived extracts used as test samples, a test sample that reduces the amount of CPD in fibroblast DNA immediately after completion of UV-B irradiation is screened as a candidate for DNA damage repair promoter. did.

As a result, a kina extract (manufactured by Maruzen Pharmaceutical Co., Ltd., trade name: Kina extract BG) was obtained as a candidate for a DNA damage repair accelerator.

(Example 1)
In the medium [Wako Pure Chemical Industries, Ltd., trade name: D-MEM], as a test sample, Kina extract [Maruzen Pharmaceutical Co., Ltd., trade name: Kina extract BG, composition of Kina extract: plant origin The extract (dry matter) 3 mass%, 1,3-butylene glycol 48.5 mass%, purified water 48.5 mass%] was added to obtain the test sample-containing medium of Example 1. In addition, it adjusted so that the density | concentration of the quina extract in a test sample containing culture medium might be 0.01 mass%, 0.1 mass%, or 1 mass%.

(Test Example 2)
In Test Example 1, the test sample-containing medium of Example 1 was used as the test sample-containing medium, and the same operation as in Test Example 1 was performed to calculate the amount of CPD.

In addition, instead of adding the test sample-containing medium to the 96-well plate containing the fibroblasts after UV-B irradiation, control the medium (trade name: D-MEM, manufactured by Wako Pure Chemical Industries, Ltd.). The same operation as described above was performed except that the medium was added as a medium, and the amount of CPD was calculated.

The CPD residual rate when a predetermined time has elapsed from the end of UV-B irradiation was determined by setting the amount of CPD in fibroblast DNA immediately after UV-B irradiation to 100.

In Test Example 2, the relationship between the elapsed time from the end of UV-B irradiation and the CPD residual rate is shown in FIG. In addition, FIG. 1 shows the result of using a test sample-containing medium whose test sample concentration is 1% by mass as a representative example. In the figure, black circles indicate the CPD residual rate of fibroblast DNA when cultured in the test sample-containing medium of Example 1, and white squares indicate the CPD residual rate of fibroblast DNA when cultured in the control medium.

From the results shown in FIG. 1, the CPD residual rate of fibroblast DNA when cultured in a control medium after 8 hours and 24 hours from the end of UV-B irradiation In comparison, it can be seen that the CPD residual rate of fibroblast DNA when cultured in the test sample-containing medium of Example 1 is low.

Next, the CPD remaining rate of fibroblast DNA immediately after UV-B irradiation when cultured in the test sample-containing medium of Example 1 and UV-B when cultured in the test sample-containing medium of Example 1 The repair rate was calculated using the CPD residual rate of fibroblast DNA after a predetermined time from the end of irradiation. In addition, the CPD residual rate of the fibroblast DNA immediately after UV-B irradiation when cultured in the control medium and the fibroblast DNA after a predetermined time from the end of UV-B irradiation when cultured in the control medium The repair rate was calculated using the residual CPD rate. Thereafter, the repair rate of damaged DNA in fibroblasts when cultured in the test sample-containing medium of Example 1 was compared with the repair rate of damaged DNA in fibroblasts when cultured in the control medium.

As a result, when fibroblasts after UV-B irradiation were cultured in a quina extract-containing medium, the rate of decrease in the amount of CPD compared to when fibroblasts after UV-B irradiation were cultured in a control medium It can be seen that is higher. Therefore, from these results, it can be seen that the kina extract is useful as a DNA repair promoter that promotes repair of damaged DNA because it has the property of promoting repair of damaged DNA.

From the above results, since the kina extract promotes the repair of damaged DNA, the DNA repair promoter containing the kina extract is useful as an ultraviolet stimulation inhibitor for improving the skin condition caused by the damaged DNA. It is suggested that

(Examples 2 to 20, Comparative Examples 1 to 3)
A plant-derived extract shown in Table 1 was added as a test sample to a medium [Wako Pure Chemical Industries, Ltd., trade name: D-MEM], and test samples of Examples 2 to 20 and Comparative Examples 1 to 3 were added. A containing medium was obtained. The composition of each plant-derived extract is shown in Table 2. In each Example and each comparative example, it adjusted so that the density | concentration of the test sample in a test sample containing culture medium might be 0.01 mass%, 0.1 mass%, or 1 mass%.

(Test Example 3)
(1) Evaluation of test sample based on amount of CPD in DNA In Test Example 1, using the test sample-containing media of Examples 1 to 20 and Comparative Examples 1 to 3, the culture time from the end of UV-B irradiation was 0 Except for the time, 8 hours or 24 hours, the same operation as in Test Example 1 was performed to calculate the amount of CPD in DNA. Further, as a control, the same operation as described above was performed except that a medium (trade name: D-MEM, manufactured by Wako Pure Chemical Industries, Ltd.) was used, and the amount of CPD in DNA was calculated.

Next, for each of the evaluation points of Experiment Nos. 1 to 9 shown in Table 3, the amount of CPD in the DNA of fibroblasts cultured in the control medium is defined as 100. The ratio of the amount of CPD in the DNA of fibroblasts cultured in the medium was calculated.

Based on the above ratio, it was evaluated whether or not the production of CPD was suppressed by the test sample. The evaluation criteria are as follows.

<Evaluation criteria>
○: The evaluation ratio is 90 or less at evaluation points of 1/3 or more of all evaluation points.
X: It does not satisfy the above evaluation criteria of ○.

Table 4 shows the results of evaluating whether or not the production of CPD is suppressed by the test sample.

(2) Evaluation of a test sample based on the amount of 6-4PP in DNA In the above (1), the amount of UV-B irradiation is the damaged DNA generated in DNA when irradiated with 160 mJ / m ² of UV-C. Instead of a 200-fold dilution of the primary antibody solution containing the anti-CPD antibody and a 6-4 photoproduct portion present in the damaged DNA in the DNA. The amount of 6-4PP in DNA was calculated in the same manner as in Test Example 1 except that a 200-fold dilution of a primary antibody solution containing an antibody (hereinafter referred to as “anti-6-4PP antibody”) was used. did.

Next, with respect to each evaluation point of Experiment Nos. 1 to 9 shown in Table 3, the amount of 6-4PP in the DNA of fibroblasts cultured in the control medium is defined as 100, and the amount of 6-4PP in the DNA of the control The ratio of the 6-4PP amount in the DNA of fibroblasts cultured in the medium containing the test sample was calculated. Based on the ratio, it was evaluated whether the production of 6-4PP was suppressed by the test sample. The evaluation criteria are as follows.

<Evaluation criteria>
○: The evaluation ratio is 90 or less at evaluation points of 1/3 or more of all evaluation points.
X: It does not satisfy the above evaluation criteria of ○.

Table 4 shows the results of evaluating whether the production of 6-4PP is suppressed by the test sample.

(3) Evaluation of the DNA damage inhibitory effect of the test sample Whether the test sample shows the effect of suppressing the production of damaged DNA (DNA damage inhibitory effect) from the evaluation result in (1) and the evaluation result in (2) Evaluated. The evaluation criteria are as follows.

<Evaluation criteria>
○: Either or both of the evaluation result in (1) and the evaluation result in (2) are ◯, and the test sample exhibits a DNA damage inhibitory effect.
X: Both the evaluation result in the above (1) and the evaluation result in the above (2) are x, and the test sample does not show a DNA damage suppressing effect.

Table 4 shows the results of evaluating whether the test sample exhibits a DNA damage inhibitory effect.

From the results shown in Table 2, it can be seen that when fibroblasts are cultured in the test sample-containing media of Examples 1 to 20, the generation of damaged DNA in the DNA of the fibroblasts is suppressed. Therefore, from these results, kina extract (Example 1), Comfrey extract (Example 2), coffee tree extract (Example 3), cuckoo extract (Example 4), burdock extract (Example 5) ), Auren extract (Example 6), Clara extract (Example 7), Chlorella extract (Example 8), Lavender extract (Example 9), Evening primrose extract (Example 10), Rose extract (Example 11), Achazul extract (Example 12), Amaranthus extract (Example 13), Adriatic extract (Example 14), Carrot extract (Example 15), Linden extract (Example 16) Since the jellyfish extract (Example 17), the lotus extract (Example 18), the tea extract (Example 19) and the okra extract (Example 20) have the property of suppressing the production of damaged DNA, DNA loss It proves useful as inhibitors. Among these, Comfrey extract, Coffea extract, Cuckoo extract, Clara extract, Chlorella extract, Lavender extract, Evening primrose extract, Rose extract, Achacha extract, Enmiso extract, Odori extract, The carrot extract, the linden extract, the jellyfish extract, the lotus extract, the tea extract and the okra extract showed high inhibitory properties against damaged DNA. Among them, comfrey extract, coffee tree extract, cuckoo extract, clara extract, chlorella extract, evening primrose extract, enmiso extract, ginger extract, lotus extract, tea extract and okra extract, these Especially, the higher suppression was shown with respect to the damage DNA by a comfrey extract, a coffee extract, a cuckoo extract, a lotus extract, and an okra extract, respectively.

Claims (2)

1. A DNA repair promoter that promotes repair of damaged DNA and contains a kina extract.
2. A DNA damage inhibitor that suppresses the generation of damaged DNA, including kina extract, comfrey extract, coffee tree extract, cuckoo extract, burdock extract, auren extract, clara extract, chlorella extract, lavender extract Selected from the group consisting of a product, an evening primrose extract, a rose extract, a candy extract, an enamel extract, a licorice extract, a carrot extract, a linden extract, a ginger extract, a lotus extract, a tea extract and a okra extract A DNA damage inhibitor comprising at least one extract.

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