JP6781925B1 - Mineral metabolism disorder inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a material capable of preventing / ameliorating spots by improving abnormal metabolism of minerals in the subepidermal region and normalizing proliferation of epidermal cells and epidermal turnover. SOLUTION: An agent for suppressing abnormal iron metabolism in the epidermis of a spot on the skin containing Houttuynia cordata as an active ingredient. [Selection diagram] Fig. 1

Description

The present invention relates to a mineral metabolism disorder inhibitor, a TFRC1 production promoter, an ACO1 production promoter, an SLC40A1 production inhibitor, a DMT1 production promoter, and an SLC25A37 production promoter.

Minerals are one of the five major nutrients along with proteins, carbohydrates, fats and vitamins, and are mainly constituents of living tissues (bones, teeth, etc.) in the body, maintenance of body fluid homeostasis (pH, osmoregulation, etc.), It plays an important role as a cofactor of enzymes, regulation of excitability of nerves and muscles, and heart, and a component of bioactive substances.

Minerals are roughly classified into two types: large elements and trace elements. Major elements include calcium, phosphorus, potassium, sodium, sulfur, magnesium and the like. Trace elements include iron, copper, zinc, selenium, manganese, iodine and the like.

That is, in order for humans to maintain their health, daily intake of various minerals is indispensable. For example, it is well known that iron deficiency causes anemia. In addition, zinc deficiency causes skin ulcers, decreased immune response, and sexual dysfunction dwarf disease, and copper deficiency causes anemia and increased plasma cholesterol concentration. On the contrary, it is known that excessive intake of iron, for example, causes skin pigmentation and liver damage, and excessive copper causes brain damage (Non-Patent Document 1).

There is a mechanism to maintain the mineral balance in the living body. For example, the sodium pump of the cell membrane consumes energy to pump out intracellular sodium ions, and there are many potassium ions inside the cells and sodium ions outside the cells. Maintains a large number of.

Of course, mineral balance is also important for maintaining skin health, and it has been reported that the presence of a large amount of magnesium ions in the stratum granulosum and calcium ions in the stratum granulosum is important for skin homeostasis (maintenance of homeostasis). Has been done. It is also known that calcium ions affect the enzyme balance related to stratum corneum exfoliation (→ keratinization), that is, the digestion of desmosomes.

Furthermore, it is known that trace amounts of minerals are localized in the epidermis. Iron, zinc, copper, etc. are all required for the activity of various enzymes involved in cell proliferation and antioxidants, and among these, iron and copper are metals that have a narrow physiological optimum concentration and are likely to lead to oxidative damage to tissues. It is known that there is. Abnormal localization of these minerals is considered to lead to abnormal skin turnover and oxidative pathology (Non-Patent Document 2).

It is generally known that epidermal turnover is initiated by the proliferation of basal cells and is responsible for the excretion of melanin.
So-called spots such as senile pigment spots are the most common spots on the face, and many people cite them as cosmetic problems. It has been reported that the epidermis is thickened and melanin is accumulated in the affected area of the spot, the expression of molecules involved in melanin production is increased, and the proliferation of basal cells is decreased.

Therefore, if the proliferation of epidermal cells in the reduced skin can be activated or the expression of molecules involved in melanin production can be reduced, the original skin function is restored, melanin is excreted by epidermal turnover, and melanin is excreted. It is considered that prevention, treatment, improvement, etc. can be realized.

Therefore, the present inventors have focused on improving mineral metabolism in the skin as a method for improving such abnormalities in epidermal turnover and suppressing the expression of molecules involved in melanin production.
This time, it was clarified that the expression of iron metabolism-related genes is altered in the subepidermal part of the pathological condition of spots, and the utilization of iron is suppressed. For example, it was found that the expression of TFRC1, ACO1, SLC25A37, and DMT1 was decreased in the subepidermal part in the blemishes pathological part. It was thought that this abnormal mineral metabolism caused an abnormal turnover of the epidermis, and melanin was not excreted to form spots.

Therefore, if the expression of molecules related to iron metabolism can be brought close to normal, that is, if a component that promotes iron utilization can be found in the subepidermal region, it is possible to restore the proliferation of epidermal cells and melanin turnover to normal. I thought that it would be possible to prevent and improve the disease.

Hiroshi Kawabata (2010) Journal of the Japan Society 99: 1173-1179. Inoue Y et al (2014) J Biol chem., 1; 289 (31): 21451-62. Asano M D et al (2017) J Dermatol Sci., 87 (2): 101-9. Kazuhiro Iwai et al. (2007) Biochemistry Vol. 79, No. 11 1021-31 Hideo Zhang (2013) Journal of Japan 102: 2699-2704

An object of the present invention is to provide a material capable of preventing and improving spots by improving the abnormal metabolism of minerals in the subepidermal region and normalizing the proliferation of epidermal cells and epidermal turnover.

As a result of diligent studies, the present inventors have an action in which specific components such as Agetsu and Scutellaria baicalensis regulate the production of proteins involved in iron uptake and excretion in cells under the epidermis and promote the utilization of iron. I found that.

That is, the present invention
(1) Agetsu, Ogon, Olive leaf, Watermelon, Star fruit leaf, Tsubokusa, Rose, Artichoke, Camomila, Perilla, Shimotsukesou, Shakuyaku, Hamamelis, Bilberry leaf, Yukinoshita, Indigo, Shokyo, Dokudami, Otogirisou, Tohi, Kiwi An inhibitor of mineral metabolism abnormalities in the subepidermal region of skin stains containing at least one selected from the group consisting of olive leaf, bilberry, tounin, lychee, and aloe as an active ingredient.
(2) The active ingredients consist of ages, honeysuckles, olive leaves, honeysuckle, star fruit leaves, tsubokusa, roses, artichokes, chamomiles, perilla, shimotsukesou, shakuyaku, hamamelis, bilberry leaves, saxifraga, indigo, shokyo, and dokudami. At least one selected from the group, the mineral metabolism abnormality inhibitor (1) based on the TFRC1 production promoting action,
(3) The active ingredient is a star fruit leaf, and the mineral metabolism abnormality inhibitor of (1) based on the ACO1 production promoting action,
(4) The active ingredient is at least one selected from the group consisting of hypericum, spruce, kiwi, dead-nettle, benibana, tounin, and lychee, and the mineral metabolism abnormality inhibitor of (1) based on the SLC40A1 production inhibitory action.
(5) The active ingredient is honeysuckle, which is a mineral metabolism disorder inhibitor of (1) based on the DMT1 production promoting action.
(6) The active ingredient is aloe, and the mineral metabolism abnormality inhibitor of (1) based on the SLC25A37 production promoting action,
(7) Selected from the group consisting of ages, honeysuckles, olive leaves, honeysuckles, star fruit leaves, honeysuckles, roses, artichokes, chamomiles, perilla, witch hazel, witch hazel, bilberry leaves, saxifraga, indigo, shokyo, and dokudami. TFRC1 production promoter containing at least one active ingredient,
(8) ACO1 production promoter containing star fruit leaves as an active ingredient,
(9) An SLC40A1 production inhibitor containing at least one selected from the group consisting of hypericum, spruce, kiwi, dead-nettle, safflower, tounin, and lychee as an active ingredient.
(10) DMT1 production promoter containing honeysuckle as an active ingredient,
(11) SLC25A37 production promoter containing aloe as an active ingredient,
(12) One or more genes selected from the group consisting of TFRC1 gene, ACO1 gene, SLC40A1 gene, DMT1 gene, and SLC25A37 when the test substance was allowed to act on undifferentiated human epidermal keratinocytes. A method for screening a stain-preventing or improving agent using the expression level as an index.

The agent for suppressing abnormal mineral metabolism of the present invention promotes the utilization of iron in cells under the epidermis by regulating the production of proteins involved in the utilization of minerals such as TFRC1, ACO1, SLC40A1, DMT1 and SLC25A37, and epidermal cells. It is possible to obtain the effect of preventing / improving spots by normalizing the proliferation and epidermal turnover. Further, through the promotion of TFRC1 production, promotion of ACO1 production or suppression of SLC40A1 production, promotion of DMT1 production, and promotion of SLC25A37 production, it is possible to obtain therapeutic / ameliorating effects on various symptoms and diseases based on excess or decrease of these proteins. Furthermore, it can also be used as a positive control agent in screening materials involved in the expression of these proteins or encoding genes. Further, according to the screening method of the present invention, components that can prevent or improve spots can be efficiently screened.

FIG. 1 is a graph showing the ΔΔCt value of the TFRC1 gene in Test Example 1. FIG. 2 is a graph showing the ΔΔCt value of the ACO1 gene in Test Example 1. FIG. 3 is a graph showing the ΔΔCt value of the SLC40A1 gene in Test Example 1. FIG. 4 is a graph showing the ΔΔCt value of the DMT1 gene in Test Example 1. FIG. 5 is a graph showing the ΔΔCt value of the SLC25A37 gene in Test Example 1.

In the present invention, the following components are used as active ingredients.

The ages used in the present invention are crude drugs or plants derived from the fruits of Rosa multiflora Thunberg or other related plants (Rosaceae) of the Rosales family Rosales, and are used in the form of crude drug powders or plant powders, crude drug extracts or plant extracts. Will be done. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The agetsu extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available products of Agetsu Extract include Maruzen Pharmaceuticals' Agetsu Extract-R, Agetsu Extract BG-R, Agetsu Extract BG-01, and Ichimaru Falcos' Falco Rex (registered trademark) Agetsu. B ”etc.

The ginger used in the present invention is a raw medicine or plant derived from the roots of Scutellaria baicalensis Georgi (Labiatae), which belongs to the genus Scutellaria baicalensis, Lamiales, in the form of raw medicine powder or plant powder, raw medicine extract or plant extract. used. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The augon extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available products of Scutellaria baicalensis include Maruzen Pharmaceuticals' Scutellaria baicalensis extract-J, Ichimaru Falcos' Scutellaria baicalensis extract powder, Scutellaria baicalensis liquid B, Scutellaria baicalensis liquid G, and Scutellaria baicalensis liquid SE. ..

The olive leaf used in the present invention is a crude drug or plant derived from the leaf of the olive genus Olea europaea of the family Oleaceae, Lamiales, and is used in the form of crude drug powder or plant powder, crude drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The olive leaf extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic alcohols. A solvent extracted from a solvent such as ketone (acetone or the like) or a mixed solution of the solvent can be used.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available olive leaf extracts include Maruzen Pharmaceuticals' Olive Leaf Extract BG.

The honeysuckle used in the present invention is a honeysuckle or plant derived from the flowers, leaves or stems of the honeysuckle Lonicera japonica Thunberg or other honeysuckle family of the order Dipsacales, honeysuckle family, honeysuckle family, honeysuckle family, honeysuckle family, honeysuckle family, honeysuckle family, honeysuckle family. Used in the form of. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The watermelon extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available products of honeysuckle extract include Maruzen Pharmaceuticals' "Kinginka Extract-J", Ichimaru Falcos' "Falcolex Honeysuckle FB", and "Falcolex Honeysuckle FE".

The star fruit leaf used in the present invention is a crude drug or plant derived from the leaf of Averrhoa carambola, which belongs to the genus Carambola of the family Oxalidales, and is used in the form of crude drug powder or plant powder, raw drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The star fruit leaf extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), lower fats. A solvent extracted from a solvent such as a group ketone (acetone or the like) or a mixed solution of the solvent can be used.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available star fruit leaf extracts include Maruzen Pharmaceuticals' Star Fruit Leaf Extract BG30 and Star Fruit Leaf Extract Powder MF.

The Centella asiatica used in the present invention is a biopharmaceutical or plant derived from the leaves and stems of Centella asiatica Linne (Umbelliferae), which belongs to the genus Centella asiatica of the Umbelliferae family, and is used in the form of a biopharmaceutical powder or plant powder, a biopharmaceutical extract or a plant extract. .. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The tuboxa extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available products of Centella asiatica extract include Maruzen Pharmaceuticals' Sekisetsuso Extract BG70.

The rose used in the present invention is a crude drug or plant derived from the flower of Rosa centifolia Linne (Rosaceae), which belongs to the genus Rosa of the Rosales family, and is used in the form of crude drug powder or plant powder, crude drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The rose extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones (such as glycerin). A solvent such as acetone) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a polyhydric alcohol, or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available rose extract products include Maruzen Pharmaceuticals' "Rose Extract BG" and Ichimaru Falcos' "Falcolex Rose P", "Rose Water", and "Rose Water PF".

The artichoke used in the present invention is a crude drug or plant derived from the leaves of Cynara scolymus Linne (Compositae), which belongs to the genus Cynara of the Asteraceae family, and is used in the form of crude drug powder or plant powder, crude drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The artichoke extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acemonate or the like) or a mixture of the same solvents can be used.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available artichoke extracts include Ichimaru Falcos's "Biobenefity (registered trademark)", "Biobenefity G", "Biobenefity HS", "Cynaropicrin F", and "Biobenefity F". ..

The chamomile used in the present invention is a crude drug or plant derived from the flower of Matricaria chamomilla Linne (Compositae) of the family Asteraceae, Asteraceae, and is used in the form of crude drug powder or plant powder, crude drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The chamomile extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone, etc.), a mixed solution of the above solvent, or a mixture of the above solvents containing urea can be used, and among them, water, lower fatty alcohol, polyhydric alcohol, or a mixed solution thereof or urea can be used. It is most preferable to extract with a mixed solution of these containing.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available chamomile extract includes Maruzen Pharmaceutical's "Chamomile Extract", "Chamomile Extract BG-J", "Chamomile Extract LA", and Ichimaru Falcos' "Ode Aroma (Registered Trademark) Chamomile", "Ode Aroma". Examples include "chamomile B", "chamomile liquid", and "biocellact (registered trademark) chamomile B".

The perilla used in the present invention is a perilla or plant derived from the leaves and branch tips of Perilla frutescens Britton Var. Acuta Kudo or other closely related perilla (Labiaatae) of the family Lamiales, Perilla frutescens Britton Var. Used in the form of perilla extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The persimmon extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.). A solvent such as acetone) or a mixture of the above solvents can be used, but water, lower fatty alcohols, polyhydric alcohols, or a mixture thereof is most preferable.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available perilla extract include Maruzen Pharmaceuticals' perilla extract BG and blue perilla extract MF (N), and Ichimaru Falcos' Falcolex perilla and Falcolex perilla HB.

The Filipendula used in the present invention is a crude drug or plant derived from the inflorescence of Filipendula ulmaria Maximowicz (Rosaceae), which belongs to the genus Filipendula of the Rosales family, and is used in the form of crude drug powder or plant powder, crude drug extract or plant extract. .. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The shimotsukesou extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available products of Filipendula multijuga extract include "Falcolex Filipendula multijuga B" by Ichimaru Falcos.

The peony used in the present invention is a peony or plant derived from the roots of Paeonia lactiflora Pallas (Paeonia albiflora Pallas var. Trichocarpa Bunge) or other closely related plants (Paeonia ceae), which belongs to the genus Peony of the order Peony, Peony. , Used in the form of peony extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The shakuyaku extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available products of peony extract include Maruzen Pharmaceuticals's peony extract-J, peony extract BG-JC, Japanese ism <peony>, and Ichimaru Falcos' peony liquid, Falcolex peony B. , Etc. can be mentioned.

The witch hazel used in the present invention is a crude drug or plant derived from the leaves or leaves and bark of Hamamelis virginiana L. (Hamamelidaceae) of the order Saxifragales, witch hazel family, witch hazel, in the form of crude drug powder or plant powder, raw drug extract or plant extract. used. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The Hamamelis extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available witch hazel extract include Maruzen Pharmaceutical's "Hamamelis Extract BG-J" and Ichimaru Falcos' "Falcolex Hamamelis B".

The bilberry leaf used in the present invention is a crude drug or plant derived from the leaf of Vaccinium myrtillus L. (Ericaceae) of the genus Vaccinium of the family Ericaceae, and is used in the form of crude drug powder or plant powder, raw drug extract or plant extract. .. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The bilberry leaf extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic alcohols. A solvent extracted from a solvent such as ketone (acetone or the like) or a mixed solution of the solvent can be used.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available products of bilberry leaf extract include "Eco Farm (registered trademark) Bilberry Leaf E", "Eco Farm Bilberry Leaf G", and "Cure Berry (registered trademark)" of Ichimaru Falcos.

The Saxifragacea used in the present invention is a saxifraga stolonifera Meerburg (Saxifragaceae), which is derived from the whole plant of Saxifraga stolonifera Meerburg (Saxifragaceae), and is used in the form of Saxifraga stolonifera Meerburg (Saxifragaceae), Saxifraga stolonifera Meerburg (Saxifragaceae). The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The Yukinoshita extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available products of Yukinoshita extract include Maruzen Pharmaceuticals' Yukinoshita extract, Yukinoshita extract BG, Japanese ism <Yukinoshita>, and Ichimaru Falcos' Falcolex Yukinoshita MB.

The indigo plant used in the present invention is a crude drug or plant derived from the leaves / stems of Polygonum tinctorium Lour. (Polygonaceae), Polygonum tinctorium Lour. (Polygonaceae), and is used in the form of raw drug powder or plant powder, raw drug extract or plant extract. To. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The indigo extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acemonate or the like) or a mixture of the same solvents can be used.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available indigo extract include "Ai Luros (registered trademark)" by Ichimaru Falcos.

The ginger used in the present invention is a raw medicine or plant derived from the rhizome of Zingiber officinale Roscoe (Zingiberaceae) of the family Zingiberaceae, Zingiberaceae, and is used in the form of raw drug powder or plant powder, raw drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The gypsum extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower fatty alcohols. A solvent extracted from a solvent such as ketone (acetone or the like) or a mixed solution of the above solvents can be used, but it is most preferable to extract with water, a lower aliphatic alcohol or a mixed solution thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available products of ginger extract include Maruzen Pharmaceuticals's "Ginger Tincture" and Ichimaru Falcos' "Falcolex Ginger E".

The Houttuynia used in the present invention is a raw medicine or plant derived from the above-ground part of the flowering period of Houttuynia cordata Thunberg (Saururaceae) of the family Houttuynia, Houttuynia, and is used in the form of raw medicine powder or plant powder, raw medicine extract or plant extract. Will be done. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The dokudami extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Commercially available products of Houttuynia cordata extract include Maruzen Pharmaceutical's "Houttuynia cordata extract", "Houttuynia cordata extract BG", "Houttuynia cordata extract powder MF", and Ichimaru Falcos "Falcolex Houttuynia cordata B", "Falcolex Houttuynia cordata E", " Falcolex Dokudami W ”,“ Dokudami DXP100 ”etc.

The Hypericum erectum used in the present invention is a raw medicine or plant derived from the above-ground part of Hypericum perforatum Linne or Hypericum erectum Thunberg (Guttiferae) of the order Hypericum erectum Thunberg (Guttiferae). Used in form. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The otogirisou extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available products of Hypericum extract include Maruzen Pharmaceuticals' "Hypericum extract BG", Ichimaru Falcos' "Falcolex Hypericum B", and "Falcolex Hypericum E".

The spruce used in the present invention is a crude drug or plant derived from the mature skin of Citrus aurantium Linne or Daidai Citrus aurantium Linne var. Daidai Makino (Rutaceae) of the Pinaceae family, and is a crude drug powder or plant powder, a crude drug extract or Used in the form of plant extracts. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The tow extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available spruce extract include Maruzen Pharmaceuticals's "Spruce extract <Glauber's salt>" and Ichimaru Falcos' "Spruce liquid" and "Spruce liquid B".

The kiwi used in the present invention is a raw medicine or plant derived from the fruit (raw) of Actinidia chinensis Planch (Actinidiaceae) of the family Actinidiaceae, Actinidiaceae, and is used in the form of raw medicine powder or plant powder, raw medicine extract or plant extract. To. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The kiwi extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent extracted with a solvent such as (acetone or the like) or a mixed solution of the above solvents can be used, but it is most preferable to extract with water.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available kiwi extracts include "Falcolex Kiwi" and "Falcolex Kiwi B" from Ichimaru Falcos.

The dead-nettle used in the present invention is a raw medicine or plant derived from the flowers, stems, and leaves of Lamium album Linne (Labiatae) of the Labiatae family Lamiales, and is used in the form of raw medicine powder or plant powder, raw medicine extract or plant extract. To. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The odorikosou extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available dead-nettle extracts include "Falcolex dead-nettle B" from Ichimaru Falcos.

The safflower used in the present invention is a crude drug or plant derived from the flower of Carthamus tinctorius Linne (Compositae), which belongs to the genus Safflower in the family Asteraceae, and is squeezed into a plate shape by removing most of the yellow pigment. It is used in the form of crude drug powder or plant powder, crude drug extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The Benibana extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available safflower extract include Maruzen Pharmaceuticals's "safflower extract" and "safflower extract BG", and Ichimaru Falcos' "Falcolex safflower B" and "safflower liquid".

The tounin used in the present invention is a biopharmaceutical or plant derived from the seeds of Prunus persica Batsch or Prunus persica Batsch var. Used in the form of. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The tounin extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower fatty ketones (such as glycerin). A solvent such as acetone) or a mixture of the above solvents can be used, but water, lower fatty alcohols, polyhydric alcohols, or a mixture thereof is most preferable.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available products of Tounin extract include Maruzen Pharmaceuticals' Tounin Extract-J and Ichimaru Falcos' Falcolex Tounin B.

The lychee used in the present invention is a raw medicine or plant derived from the fruit of Litchi chinensis Sonn. (Sapindaceae) of the family Sapindales, Sapindales, and is used in the form of raw medicine powder or plant powder, raw medicine extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The lychee extract used in the present invention includes water, lower aliphatic alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acemonate or the like) or a mixture of the same solvents can be used.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available lychee extracts include Ichimaru Falcos's "ADS-Oligonol" and "Oligonol-CS".

The aloe used in the present invention is Aloe Ferox Miller or a hybrid of Aloe Africana Miller or Aloe spicata Baker (Liliaceae) or Aloe barbadensis Miller (Aloe vera Linne) or Aloe arborescens Miller and its variants. A herb or plant derived from the leaves or juice of (Liliaceae), used in the form of aloe powder or plant powder, aloe extract or plant extract. The crude drug powder or plant powder used in the present invention may be, for example, a powdered dried product obtained by further finely crushing a dried chopped product.

The aloe extract used in the present invention includes water, lower fatty alcohols (methanol, ethanol, isopropyl alcohol, etc.), polyhydric alcohols (1,3-butylene glycol, propylene glycol, dipropylene glycol, glycerin, etc.), and lower aliphatic ketones. A solvent such as (acetone or the like) or a mixture of the above solvents can be used, but it is most preferable to extract with water, a lower fatty alcohol, a polyhydric alcohol or a mixture thereof.

The form of the extract is not particularly limited, and dried extract powder, extract powder, soft extract, stream extract and the like can be obtained by heat treatment, freeze-drying or vacuum drying. Examples of commercially available extracts include and the like. Examples of commercially available aloe extract products include "Arrow Cable Liquid" and "Arrow Cable Liquid B-7 (N)" from Ichimaru Falcos.

The skin is roughly divided into three layers: epidermis, dermis, and subcutaneous tissue. The major constituent cells of the epidermis are keratinocytes (keratinocytes), which show four morphological changes in the process of differentiation. The basal cells in the lowest layer are in contact with the basement membrane, perform DNA synthesis, and divide and proliferate. The divided cells move upward and become stabbed cells. As the differentiation progresses, it becomes granule cells, and finally it differentiates from granule cells to stratum corneum cells to form the uppermost stratum corneum. Keratinocytes at each of these differentiation stages are stacked in the epidermis. The stratum corneum cells that form the stratum corneum are adhered to each other by desquamation, but in healthy skin, the adhesion mechanism gradually declines, and the stratum corneum naturally detaches due to physical stimulation. (Exfoliation of the stratum corneum). In this way, the epidermis repeats proliferation, differentiation, and exfoliation at a constant rhythm (epidermis turnover). In addition to keratinocytes, melanin cells are present in the epidermis and produce melanin pigment. Normally, melanin pigment is transferred to keratinocytes and excreted during epidermal turnover with exfoliation of the stratum corneum. .. However, when abnormalities occur in epidermal turnover such as decreased differentiation / proliferation from basal cells or hyperproliferation of keratinocytes resulting in stratification / thickening, melanin pigment is not excreted and accumulates in the epidermis to cause spots. Is formed. In the present specification, the subcutaneous part of the epidermis means a layer containing basal cells among the keratinocytes constituting the epidermis, and the upper part of the epidermis means a layer containing keratinocytes.

The transferrin receptor (TFRC1) is expressed in various cells in the living body, and after iron in serum is bound to transferrin and transported, iron is taken up into the cells via TFRC1. When cells lack iron, the expression of TFRC1 decreases, and thus plays an important role in the uptake of iron into cells. When TFRC1 production is promoted in keratinocytes under the epidermis, the uptake of iron into the cells is promoted, the intracellular iron concentration is increased, epidermal turnover and epidermal layer regeneration are carried out, and melanin pigment is used. Since the discharge is promoted, the effect of preventing and / or improving the stain can be obtained (Non-Patent Documents 1 and 3).

Of the above active ingredients, witch hazel, honeysuckle, olive leaf, honeysuckle, star fruit leaf, centella asiatica, rose, artichoke, chamomile, perilla, witch hazel, bilberry leaf, saxifraga, indigo, shokyo, and dokudami are subepithelial. TFRC1 production promoting action in.

ACO1 is an enzyme that functions as an essential enzyme in the TCA cycle and catalyzes the reaction of citric acid to isocitric acid through aconitate. When the intracellular iron concentration is high, this protein functions as an aconitase. When cells have low iron levels, proteins bind to the iron response element (IRE). When ACO1 production is promoted in keratinocytes under the epidermis, mitochondria of basal cells are activated, and iron utilization is normalized by the stabilizing action of intracellular iron concentration, and epidermal turnover is promoted. Since the excretion of melanin is also promoted, the effect of preventing and / or improving stains can be obtained (Non-Patent Document 4).

Among the above active ingredients, star fruit leaves have an ACO1 production promoting action.

SLC40A1 (ferroportin) is responsible for the excretion of iron from cells. The expression of ferroportin is regulated by intracellular iron excess and deficiency and by hepcidin according to individual iron excess and deficiency at the protein level. Ferroportin is known to be a protein that excretes iron from intestinal epithelial cells and macrophages into blood, but in recent years, it has been reported that it is also expressed in skin tissues. Suppression of SLC40A1 production in subepidermal keratinocytes suppresses the decrease in intracellular iron concentration of keratinocytes, thereby normalizing cell proliferation and epidermal turnover, preventing spots and / Alternatively, an improvement effect can be obtained (Non-Patent Documents 1, 3 and 5).

Among the above active ingredients, Hypericum erectum, Spruce, Kiwi, White dead-nettle, Safflower, Ferroportin, and Lychee have an SLC40A1 production inhibitory effect.

DMT1, one of the divalent metal transporters, is responsible for the uptake of iron from the intestinal tract and the transport of iron from endosomes. DMT1 binds to various divalent metals, including cadmium (Cd 2+), copper (Cu 2+), and zinc (Zn 2+,), but is best known for its role in the transport of iron (Fe 2+). Has been done. Expression of DMT1 is regulated by iron stores in the body to maintain iron homeostasis. In the gastrointestinal tract, it is located in the apical membrane of enterocytes and is known to bind and transport divalent metal cations from the intestinal cavity to cells. In recent years, it has been reported that it is also expressed in the epidermis of skin tissues. When DMT1 production is promoted in keratinocytes under the epidermis, iron uptake into the cells is promoted, the intracellular iron concentration is increased, and epidermal turnover and epidermal layer regeneration are normalized. The effect of preventing and / or improving stains can be obtained.

Among the above active ingredients, honeysuckle has a DMT1 production promoting action.

In SLC25A37 (Mitoferrin1), mitochondria are the main intracellular iron-utilizing organelles, and function as an iron uptake transporter in the inner mitochondrial membrane. It exists as a major molecule of iron metabolism in intracellular mitochondria. When SLC25A37 production is promoted in keratinocytes under the epidermis, intracellular iron concentration is increased based on the promotion of iron transport to intracellular mitochondria, and epidermal turnover and epidermal layer regeneration are normalized. The prevention and / or improvement effect of the above can be obtained (Non-Patent Document 5).

Of the above active ingredients, aloe has an SLC25A37 production promoting action.

As described above, the present invention includes ages, corns, olive leaves, watermelon leaves, star fruit leaves, tsubokusa, roses, artichokes, chamomiles, perilla, ferroportin, shakuyaku, hamamelis, bilberry leaves, yukinoshita, indigo, shokyo, and TFRC1 production promoting action by at least one species selected from the group consisting of Dokudami, ACO1 production promoting action by star fruit leaves, or at least one species selected from the group consisting of bilberry, touhi, kiwi, odorikosou, benibana, tounin, and lychee. SLC40A1 production promoting action by SLC40A1, DMT1 production promoting action by watermelon, SLC25A37 production promoting action by Aloe, mineral metabolism abnormality inhibitor in the subepithelial part of the skin spot, TFRC1 production promoting agent by each of the above active ingredients, ACO1 production promoting agent, SLC40A1 production inhibitor, DMT1 production promoter, SLC25A37 production promoter are included, and further, preventive / ameliorating effect of diseases / symptoms caused by reduction of TFRC1, reduction of ACO1, excess of SLC40A1, reduction of DMT1, reduction of SLC25A37. Has.

The mineral metabolism disorder inhibitor, TFRC1 production promoter, ACO1 production promoter, SLC40A1 production inhibitor, DMT1 production promoter, SLC25A37 production promoter of the present invention are provided as cosmetics, quasi-drugs, pharmaceuticals, foods and drinks, etc. be able to. Although it is preferably applied to humans, it can also be applied to animals other than humans as long as the effects are exhibited.

The mineral metabolism disorder inhibitor, TFRC1 production promoter, ACO1 production promoter, SLC40A1 production inhibitor, DMT1 production promoter, SLC25A37 production promoter of the present invention can be used, for example, for the study of related symptoms and diseases, or for stains. It can also be used as a reagent for normalizing abnormal mineral metabolism at a site, promoting TFRC1 production, promoting ACO1 production, suppressing SLC40A1 production, promoting DMT1 production, and promoting SLC25A37 production, which is suitable. Can be used as a positive control drug when performing material screening and the like. The present invention includes a method for promoting TFRC1 production, ACO1 production, DMT1 production, SLC25A37 production, and a method for suppressing SLC40A1 production in such tests and studies. As a method for promoting TFRC1 production, epidermal keratinocytes or cells for which TFRC1 production is desired to be promoted include Hamamelis, Drosophila, Rose, Yukinoshita, Agetsu, Watermelon, Artichoke, Indigo, Shimotsukesou, Starfruit leaf, Shakuyaku, Bilberry leaf. , Camomila, Ogon, Dokudami, Shiso, Olive leaf, Tsubokusa or each extract. The method for promoting ACO1 production includes a step of adding star fruit leaves or an extract thereof to epidermal keratinocytes or cells for which ACO1 production is desired to be promoted. The method for promoting DMT1 production includes a step of adding honeysuckle or an extract thereof to epidermal keratinocytes or cells for which DMT1 production is desired to be promoted. The method for promoting SLC25A37 production includes a step of adding aloe or an extract thereof to epidermal keratinocytes or cells for which SLC25A37 production is desired to be promoted. The method of suppressing SLC40A1 production includes a step of adding ferroportin, spruce, kiwi, ferroportin, ferroportin, lychee or an extract thereof to epidermal keratinocytes or cells for which SLC40A1 promotion is desired to be suppressed.

The administration form is not particularly limited, but oral administration is preferable. Dosage forms when applied orally include tablets, powders, powders, granules, liquids, capsules, dry syrups, jellies, liquid foods, semi-solid foods, solid foods and the like. These can be produced by known methods. In the production, various additives that can be contained in quasi-drugs, pharmaceuticals, foods and drinks, or reagents can be blended as long as the effects of the present invention are not impaired.

The blending amount of the above active ingredient in the present invention is not particularly limited, but when provided as cosmetics, quasi-drugs, pharmaceuticals, foods and drinks or reagents, 0.000001 to 10% by mass with respect to the entire composition, respectively. %, preferably 0.0001 to 5% by mass, more preferably 0.001 to 1% by mass.

Mineral metabolism disorder inhibitor, TFRC1 production promoter, ACO1 production promoter, SLC40A1 production inhibitor, DMT1 production promoter, SLC25A37 production promoter (pharmaceutical products, quasi-drugs, foods and drinks, reagents, etc.) and their description thereof. The book is an indication that it is used to promote TFRC1 production, an indication that it is used to promote ACO1 production, an indication that it is used to suppress SLC40A1 production, and an indication that it is used to promote DMT1 production. , SLC25A37 may be labeled as being used for promoting production. Here, "labeled" means a product body, a container, and a product containing a mineral metabolism abnormality inhibitor, TFRC1 production promoter, ACO1 production promoter, SLC40A1 production inhibitor, DMT1 production promoter, and SLC25A37 production promoter. Describe on packaging, etc., or on documents such as manuals, attachments, pamphlets, and other printed matter that disclose product information, and on various leaflets and advertisements used for advertising including the Internet. including.

The screening method of the present invention uses undifferentiated human epidermal keratinocytes. The medium used for cell culture is not particularly limited, known ones can be used, and the cells can be cultured under normal culture conditions. The test substance was added to the medium of undifferentiated human epidermal keratinized cells cultured in this manner, and after culturing for a predetermined time, it was selected from the group consisting of the TFRC1 gene, ACO1 gene, SLC40A1 gene, DMT1 gene, and SLC25A37. The expression level of one or more genes is measured. The expression level of the gene is not particularly limited, and a known method can be used. The untreated group to which the test substance is not added is also cultured in the same manner, the expression level of each gene is measured, and compared with the expression level of each gene in the test substance treated group, thereby preventing / improving the stain of the test substance. Can be determined. In addition, for example, TFCR1, DMT1, and SLC40A1 (ferroportin; ferroportin) are tested based on the expression levels of these genes in the blemishes pathological model because the expression levels in the subepidermal region of the blemishes pathological site are decreased. By comparing the gene expression levels when the substance is added, it is possible to determine whether or not the test substance has a stain-preventing / ameliorating effect. In addition, the same test was performed using differentiated human epidermal keratinocytes, and a test substance that increased the expression of TFCR1 in undifferentiated cells but did not increase the expression of TFCR1 in the differentiated cells was selected. By combining the index of the amount of change in the specific gene expression in the pathological part of the stain, it becomes possible to select a component having a more excellent effect of preventing / improving the stain.

The present invention will be described in more detail with reference to Examples and Test Examples, but the present invention is not limited to the Examples.

(Test Example 1) Evaluation of promotion or suppression of gene expression in undifferentiated human epidermal keratinocytes <Test method>
Human epidermal keratinocytes (the Kurabo) were seeded at a density of 3.8 × ¹⁰ 4 cells / well in 12-well plates, 37 ° C., and cultured in an incubator set at _CO 2 5%. Humedia-KG2 (Kurabo Industries Ltd.) was used as the medium. The day after sowing, the medium was replaced with a medium containing the test substance, and the cells were cultured in a CO ₂ incubator for 1 day. Then, lysate buffer was added to lyse the cells, and the cell lysate was collected. RNA was recovered from the cell lysate according to the attached protocol using RNeasy Mini Kit (Qiagen), and cDNA was synthesized by reverse transcription reaction using Prime Script RT Master mix (Takara Bio) using this as a template. From the synthesized cDNA, the expression levels of RPLP0, TFRC1, ACO1, SLC40A1, DMT1, and SLC25A37 mRNAs were measured by the SYBR Green method using a real-time PCR system (Step One Plus, Thermo Fisher Scientific). The Ct value of each sample of TFRC1, ACO1, SLC40A1, DMT1 and SLC25A37 was subtracted from the Ct value of the internal standard gene RPLP0 to obtain the ΔCt value. Further, the value obtained by subtracting the ΔCt value of the sample in the untreated group from the ΔCt value of the sample in the test substance-added group was taken as the ΔΔCt value. For example, if the ΔΔCt value is -1, it means that the gene expression is doubled, and if it is 1, it means that the gene expression is halved.
As the primer, the one having the following sequence or model number was used. RPLP0: (F) 5'-GAAGCCACGCTGCTGAACA-3'(R) 5'-CTGGCAACATTGCGGACA-3', SLC25A37: (F) 5'-GAAGCAGCACATTTGTCATTCA-3'(R) 5'-CTGGCAACATTGCA-3' : HA277353, ACO1: HA210761, SLC40A1: HA161921, DMT1: HA142595, (Takara Bio).
All test substances were used at a final concentration of 1% except for 0.01% lychee. The untreated group was cultured in a medium without adding the test substance, and all other conditions were the same as those in the test substance-added group.

<Test results>
FIG. 1 is a graph showing the ΔΔCt value of the TFRC1 gene. Compared to the untreated group, Agetsu, Ogon, Olive leaf, Honeysuckle, Star fruit leaf, Tsubokusa, Rose, Artichoke, Camomila, Perilla, Shimotsukesou, Shakuyaku, Hamamelis, Bilberry leaf, Yukinoshita, Indigo, Shokyo, Dokudami are TFRC1 Increased gene expression.
FIG. 2 is a graph showing the ΔΔCt value of the ACO1 gene. Star fruit leaves and ACO1 gene expression were increased as compared to the untreated group.
FIG. 3 is a graph showing the ΔΔCt value of the SLC40A1 gene. Hypericum, dead-nettle, kiwi, tounin, spruce, lychee, and safflower reduced SLC40A1 gene expression compared to the untreated group.
FIG. 4 is a graph showing the ΔΔCt value of the DMT1 gene. Honeysuckles increased DMT1 gene expression compared to the untreated group.
FIG. 5 is a graph showing the ΔΔCt value of the SLC25A37 gene. Aloe increased SLC25A37 gene expression compared to the untreated group.

The agent for suppressing abnormal mineral metabolism in the subepithelial region of the spot portion of the present invention has a stain prevention / ameliorating effect based on the TFRC1 production promoting action, ACO1 production promoting action, SLC40A1 production suppressing, DMT1 production promoting, and SLC25A37 production promoting action. It can be used in the fields of quasi-drugs, pharmaceuticals or foods and drinks. Furthermore, the TFRC1 production promoter, ACO1 production promoter, SLC40A1 production inhibitor, DMT1 production promoter, and SLC25A37 production promoter of the present invention can be used as positive control agents in material screening and the like.

Claims (3)

An agent that suppresses abnormal iron metabolism in the epidermis of the spots on the skin containing Houttuynia cordata as an active ingredient. The iron metabolism disorder inhibitor according to claim 1, which is based on the TFRC1 production promoting action. A TFRC1 production promoter containing Houttuynia cordata as an active ingredient.