JP2008273875A - Agent for suppressing increase of expression of stem cell factor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an agent for suppressing the increase of expression of a stem cell factor (SCF) by finding a substance having an action to suppress the increase of expression of a stem cell factor (SCF) from natural materials having high safety and using the substance as an active component of the agent, and to provide a prevention or treatment agent containing the suppressing agent. <P>SOLUTION: The agent for suppressing the increase of expression of SCF comprises one or more vegetable extracts selected from the extracts of Averrhoa carambola L., Glychyrrhiza glabra, Rubus ellipticus, Eriobotrya japonica Lindley, Machilus odoratissima, Melissa officinalis, Alpinia spesiosa, Coix lachrymal-jobi L., Ligusticum wallichii Franch., Prunus persica seed and Gentiana Lutea, as an active component, and the preventing or treating agent contains the suppressing agent. The suppressing agent suppresses the abnormal growth of myeloblast and is useful for the prevention or treatment of myelodysplastic syndromes, acute myelocytic leukemia, etc. The suppressing agent is also useful as a cosmetic effective for the prevention or suppression of pigmentation, spot, freckle, etc., after sunburn by suppressing the growth of pigment cell and the excessive production of melanin in the skin. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a stem cell growth factor expression increase inhibitor.

  Spots, buckwheat, skin pigmentation after sunburn, etc. occur as a result of markedly increased melanin production due to activation of pigment cells (melanocytes) present in the skin. It is connected. Conventionally, various therapeutic agents, external preparations for skin, cosmetics and the like are known as having a whitening effect. For example, as a compound that inhibits tyrosinase activity and suppresses melanin production, sulfur compounds represented by colloidal sulfur and glutathione are used. In addition, ascorbic acids, hydrogen peroxide, hydroquinone, catechol and the like are used to lightly bleach the produced melanin.

  However, ascorbic acids are unstable because they are easily oxidized when they are contained in water-rich cosmetics such as water-containing cosmetics, and cause discoloration of the cosmetics. Further, the hydrogen peroxide solution has problems in storage stability and safety in use. Furthermore, sulfur compounds such as glutathione and colloidal sulfur have a problem in that they emit a significant off-flavor. Furthermore, hydroquinone and catechol have skin irritation and allergic properties and have a problem in safety in use. Therefore, the use of these whitening ingredients in products such as cosmetics is restricted, and no whitening ingredients that are sufficiently satisfactory are yet known, and the development of better whitening agents is desired.

  Stem Cell Factor (SCF), also called Must Cell Growth Factor, C-Kit Ligand, Steel Factor, etc., is a protein produced from keratinocytes, fibroblasts, vascular endothelial cells, bone marrow stromal cells, etc. It is. SCF is known to have an action of promoting proliferation and differentiation of pluripotent hematopoietic stem cells, germ cells, mast cells, megakaryocyte progenitor cells, granulocyte / macrophage progenitor cells, pigment cells and the like. Moreover, it is known that the expression of SCF is enhanced by a spot site, ultraviolet irradiation or the like (see Non-Patent Document 1).

  As SCF, a membrane-bound SCF composed of 273 amino acid residues and a secreted SCF that is cleaved by the action of a proteolytic enzyme and released from the membrane are known. Membrane-bound SCF binds to the SCF receptor of the pigment cell while bound to keratinocytes and promotes the proliferation of the pigment cell. Secreted SCF is cleaved at its binding site, released from the cell membrane, and promotes proliferation of pigment cells by binding to SCF receptors of pigment cells.

  In addition, SCF is a myeloblast in the presence of interleukin-3 (IL-3) and granulocyte macrophage colony stimulating factor (GM-CSF) in patients with acute myeloid leukemia. It is known to promote the growth of selenium (see Non-Patent Document 2).

  Therefore, abnormal production of SCF is thought to lead to abnormal growth of pigment cells, increase melanin production, and cause stains, buckwheat, dullness, and the like. Abnormal production of SCF is thought to lead to abnormal growth of myeloblasts, thereby causing diseases such as myelodysplastic syndrome and acute myeloid leukemia (AML).

Therefore, suppressing the increase in SCF expression suppresses the proliferation of pigment cells, suppresses the excessive production of melanin in the skin, and is considered useful for the prevention or suppression of pigmentation, sun spots, buckwheat, etc. after sunburn. It is done. Further, suppressing the increase in the expression of SCF suppresses abnormal proliferation of myeloblasts and is considered useful for the prevention or treatment of myelodysplastic syndrome, acute myeloid leukemia and the like.
Hachiya A et al., J. Invest. Dermatol., No.116, 2001, p.578-586 Virginia C. Broudy et al., Blood, Vol.80, No.1, 1992, p.60-67

  An object of the present invention is to provide a highly safe natural product having an inhibitory effect on the increase in SCF expression, and to provide an agent for suppressing an increase in the expression of stem cell growth factor (SCF) containing the active ingredient as an active ingredient.

  In order to solve the above-mentioned problems, the stem cell growth factor (SCF) expression increase inhibitor of the present invention is a pentagonal extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract. It is characterized by containing, as an active ingredient, one or more plant extracts selected from the group consisting of moon peach extract, pearl barley extract, nematode extract, tonin extract and gentian extract.

  In addition, the preventive / therapeutic agent for diseases caused by the increased expression of stem cell growth factor (SCF) of the present invention is a quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa One or more plant extracts selected from the group consisting of an extract, moon peach extract, pearl barley extract, nematode extract, tonin extract and gentian extract are contained as active ingredients .

  According to the present invention, quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, nematode extract, tonin extract And one or more plant extracts selected from the group consisting of gentian extracts as active ingredients, and an excellent inhibitor of stem cell growth factor (SCF) expression increase and stem cell growth factor (SCF) A prophylactic / therapeutic agent for diseases caused by increased expression can be provided.

Hereinafter, the present invention will be described in detail.
[Suppressor of stem cell growth factor expression increase]
The stem cell growth factor (hereinafter referred to as “SCF”) expression-inhibiting agent of the present invention includes five coconut extracts, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach One or two or more kinds of plant extracts selected from the group consisting of an extract, pearl barley extract, sensu extract, tonin extract and gentian extract are contained as active ingredients.

  Here, in the present invention, “extract” refers to an extract obtained from the above plant as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude product thereof. Both purified products and purified products are included.

  The extraction raw materials used in the present invention are pentagon, licorice leaf part, oni strawberry, loquat, matils, melissa, moon peach, pearl barley, senkyu, tonin and gentian. When using two or more kinds of plants as an extraction raw material, the above plants can be used in any combination.

  Goshiko (scientific name: Averrhoa carambola L.) belongs to the genus Carenaceae, and fresh fruits are edible. Goshiko has been described in literature in China since the BC, and its fruit is also called a star fruit because of its star-shaped cross section. Gosho is cultivated in Okinawa, southeastern China, Yunnan and other tropical regions, and can be easily obtained from these regions. Examples of the part that can be used as an extraction raw material include a leaf part, a stem part, an above-ground part, a flower part, a fruit part, a seed part, a root part, a whole plant, or a mixture of these parts. is there.

  Licorice belongs to the genus Lizoaceae and is a useful plant that has been used as a raw material for medicinal or sweeteners since ancient times. In particular, it has been confirmed that the root part of licorice contains glycyrrhizin and other useful ingredients, but these useful ingredients are not contained in the leaves of licorice. On the other hand, the component which has the SCF expression rise inhibitory effect confirmed for the first time by this inventor is contained in the licorice leaf part.

  Licorice includes Glychyrrhiza glabra, Glychyrrhiza inflata, Glychyrrhiza uralensis, Glychyrrhiza aspera, Glychyrrhiza eurycarpa, Glychyrrhiza pallidiflora, Glychyrrhiza yunnanensis, Glychyrrhiza lepidchy, Glychyrrhly The licorice leaf part may be used as the extraction raw material, but it is particularly preferable to use the licorice leaf part of Glychyrrhiza glabra as the extraction raw material.

  Oni strawberry (scientific name: Rubus ellipticus, Chinese name: Kurikage Kyoko, aka: Kimino Himalayan strawberry) is an evergreen shrub belonging to the genus Rosaiaceae, and its fresh fruit is fragrant and edible. Oni strawberry is native or cultivated in the Southeast Asia and Southeast China from the Himalayas, and can be easily obtained from these regions. Examples of a part that can be used as an extraction raw material include a leaf part, a stem part, an above-ground part, a flower part, a fruit part, a seed part, a root part, a whole plant, or a mixture of these parts, but preferably a root part. .

  Biwa (scientific name: Eriobotrya japonica Lindley) is an evergreen tree belonging to the genus Biwa of the family Rosaceae, native to the southwestern part of China, and cultivated in Nagasaki, Chiba, Kagoshima, etc., and can be easily obtained from these areas it can. Examples of the part that can be used as the extraction raw material include a leaf part, a trunk part, a branch part, a flower part, a fruit part, a seed part, a root part, or a mixture of these parts, and a leaf part is preferable. The loquat of loquat is listed in the “Special Medical Supplement” and has long been used as a medicinal herb. It is said that the name is given because the shape of the leaf resembles a camellia. In Japan, in the Edo period, the decoction that was made by removing the loquat leaves and drying them was called “Yakuyaku-yu” and was used to remove heat.

  Machilus (scientific name: Machilus odoratissima) is an evergreen tree belonging to the genus Machilus, which is distributed in Yunnan, Guizhou, Sichuan, Tibet, etc., and can be easily obtained from these areas. Examples of the part that can be used as the raw material for extraction include bark part, leaf part, stem part, above-ground part, flower part, fruit part, seed part, root part, whole grass or a mixture of these parts, but preferably Bark.

  Melissa (scientific name: Melissa officinalis) is a perennial herb belonging to the genus Kosuihakka, and is also known as Kousuihakka, lemon balm, etc. Melissa is native or cultivated in the Mediterranean coast. As the word Melissa comes from the Greek words melissophyllon and bee leaf, it has been cultivated as a beekeeping plant for over 2000 years and has antiallergic effects. It is also known. Examples of a part that can be used as an extraction raw material include a leaf part, a stem part, an above-ground part, a flower part, a fruit part, a seed part, a root part, a whole plant, or a mixture of these parts. is there.

  Tsuki-Peach (scientific name: Alpinia spesiosa) is a perennial evergreen herb belonging to the genus Glyceraceae, and is distributed from southern Kyushu to India, and can be easily obtained from these regions. Tsuki-Peach is called Sannin in Okinawa and is used in Muchy, a traditional confectionery since the Ryukyu dynasty. Examples of the part that can be used as an extraction raw material include a leaf part, a stem part, an above-ground part, a flower part, a fruit part, a seed part, a root part, a whole plant, or a mixture of these parts. is there.

  The pearl barley (scientific name: Coix lachrymal-jobi L.) is a plant native to the Indochina region of China, and is an annual plant belonging to the genus Jusdama, which is cultivated in the warm southwestern part of Japan. Can be easily obtained from Examples of constituent parts that can be used as an extraction raw material include leaves, branches, bark parts, trunks, stem parts, fruit parts, seed parts, flower parts, root parts, or a mixture of these parts, but preferably Is the seed part. The seed part of pearl barley is particularly called Yokuinin, and is used for purposes such as wet water wetting, clean fever, drainage, numbness, and splenic pain.

  Senkyu (scientific name: Ligusticum wallichii Franch.) Is a plant native to China and is a perennial plant belonging to the genus Marbatoki which is cultivated in Hokkaido, Tohoku, Nagano, Shizuoka, Mie etc. in Japan. Can be easily obtained from Examples of constituent parts that can be used as the raw material for extraction include leaves, branches, bark parts, trunks, stem parts, fruit parts, seed parts, flower parts, rhizome parts, root parts, or a mixture of these parts. Is preferably a rhizome part (excluding the root part). The rhizome part (excluding the root part) of Senkyu is mainly used as a herbal medicine, and is formulated as a feminine medicine, a cold medicine, a skin disease medicine, and an anti-inflammatory back pus medicine.

  Tounin is a seed of peach (scientific name: Prunus persica) belonging to the genus Rosaceae, and has been cultivated for a long time in China and Japan and can be easily obtained from these regions. Many varieties of peach are differentiated, flowers are used for ornamental purposes, and fruits are used for food. Tonin has been used as a folk medicine for a long time, and is known to have blood purification, antitussive, anti-inflammatory and other actions.

  Gentiana (scientific name: Gentiana Lutea) is a perennial herb belonging to the genus Gentianaceae, reaching a height of more than 1 m and growing in the mountains. Gentian is native to the foothills of the Alps in Europe and is readily available from these regions. Examples of constituent parts that can be used as the raw material for extraction include leaves, branches, bark parts, trunks, stem parts, fruit parts, seed parts, flower parts, rhizome parts, root parts, or a mixture of these parts. Are preferably roots and rhizomes.

  Contained in quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, sensu extract, tonin extract or gentian extract Although the details of the SCF expression-increasing inhibitory substance are not known, depending on the extraction method generally used for the extraction of plants, five eggplants, licorice leaf part, oni strawberry, loquat, matils, melissa, moon peach, An extract having an inhibitory effect on the increase in SCF expression can be obtained from pearl barley, nematode, tonin and gentian.

  For example, after drying the plant, it is pulverized as it is or using a crusher, and subjected to extraction with an extraction solvent, whereby an extract having an SCF expression increase-inhibiting action can be obtained. Drying may be performed in the sun or may be performed using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction treatment with the polar solvent of the plant can be efficiently performed.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These may be used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. Is preferred.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a liquid mixture of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by weight of a lower aliphatic alcohol with respect to 10 parts by weight of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by mass of a lower aliphatic ketone with 10 parts by mass of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10-90 mass parts of polyhydric alcohol with respect to a mass part.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in 5 to 15 times the extraction solvent (mass ratio) of the extraction raw material, the soluble components are eluted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like. The obtained extract can be used as it is as an active ingredient of the SCF expression increase inhibitor, but a concentrated solution or a dried product is easier to use.

  Five coconut extract, licorice leaf extract, oni strawberry extract, loquat extract, matilus extract, melissa extract, moon peach extract, pearl barley extract, sensu extract, tonin extract or gentian extract Since it has a unique odor and taste, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. Is not used in large quantities, so there is no practical problem even if it is not purified.

  The quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, nematode extract, tonin extract extracted as described above Since the product or the gentian extract has an SCF expression increase inhibitory action, it can be used as an active ingredient of an SCF expression increase inhibitor using the action.

  Also, quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, barley extract, sensu extract, tonin extract or gentian extract Is a prophylactic / therapeutic agent for diseases caused by increased SCF expression, specifically, a myelodysplastic syndrome preventive / therapeutic agent, an acute myeloid leukemia prophylactic / therapeutic agent, It can also be used as an active ingredient such as a tumor agent.

  The SCF expression increase inhibitor or bFGF expression increase inhibitor of the present invention includes five coconut extracts, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley It may consist of only one or two or more plant extracts selected from the group consisting of an extract, a nematode extract, a tonin extract and a gentian extract, or a quince extract, a licorice leaf extract, One or more plants selected from the group consisting of oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, nematode extract, tonin extract and gentian extract An extract may be formulated.

  Consists of five lion extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, sensu extract, tonin extract and gentian extract One or more plant extracts selected from the group are in the form of powder, granules, tablets in accordance with a conventional method using a pharmaceutically acceptable carrier such as dextrin and cyclodextrin and other optional auxiliaries. In addition, it can be formulated and provided in an arbitrary dosage form such as liquid. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent and the like can be used. Also, quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, barley extract, sensu extract, tonin extract and gentian extract One or more plant extracts selected from the group consisting of can be used in combination with other compositions, and can also be used as an ointment, an external liquid, a patch, and the like.

  It should be noted that the SCF expression increase inhibitor of the present invention includes, as necessary, other natural extracts having an SCF expression increase inhibitory effect, such as a quince extract, licorice leaf part extract, oni strawberry extract, and loquat extract. , Effective in combination with one or more plant extracts selected from the group consisting of: Matils extract, Melissa extract, Moon peach extract, pearl barley extract, Sensu extract, Tonin extract and Gentian extract It can be used as a component.

  Examples of the method for administering the SCF expression increase inhibitor of the present invention to a patient include subcutaneous tissue administration, intramuscular administration, intravenous administration, oral administration, transdermal administration, and the like. -A method suitable for treatment or the like may be appropriately selected. The dose of the SCF expression increase inhibitor of the present invention may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.

  The SCF expression increase inhibitor of the present invention is a quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, senkyu extract The increase in SCF expression can be suppressed through the SCF expression increase-inhibiting action of one or more plant extracts selected from the group consisting of Tonin extract and Gentian extract. Proliferation and production of melanin can be suppressed, spots, buckwheat, skin pigmentation and the like can be prevented or improved, and a whitening effect can be obtained.

  Also, quince extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, barley extract, sensu extract, tonin extract and gentian extract The abnormal growth of myeloblasts can be suppressed through the SCF expression increase inhibitory action of one or more plant extracts selected from the group consisting of: myelodysplastic syndrome, acute myeloid leukemia, etc. Can be prevented, treated or ameliorated.

  However, the SCF expression increase inhibitor of the present invention can be used for all purposes that are meaningful for exhibiting the SCF expression increase suppression action in addition to these applications.

[Skin cosmetic]
Penta extract of the present invention, licorice leaf extract, oni strawberry extract, loquat extract, matilus extract, melissa extract, moon peach extract, barley extract, sensu extract, tonin extract and gentian extract An SCF expression increase inhibitor containing one or more plant extracts selected from the group consisting of substances as an active ingredient can be used by blending it into a skin cosmetic.

  The blending amount of the SCF expression increase inhibitor in skin cosmetics can be adjusted as appropriate depending on the type of skin cosmetics, physiological activity of the extract, etc., but a suitable blending rate is 0 in terms of a standard extract. 0.001 to 10% by mass.

  The skin cosmetics that can be blended with the SCF expression increase inhibitor of the present invention are various main ingredients that are usually used in the production of skin cosmetics as needed, as long as the effects of the SCF expression increase inhibitor of the present invention are not impaired. And auxiliary agents can be used in combination.

  Examples of components that can be used in combination with the SCF expression increase inhibitor of the present invention as a skin cosmetic component include glycerin, collagen, hyaluronic acid and its salts, chondroitinic acid and its salts, chitin, chitosan and other moisturizing agents; UV absorbers such as amyl aminobenzoate; complex lipids such as glycerophospholipid and sphingophospholipid; β-carotene, oil-soluble licorice extract, lycochalcone A, baicalin, baicalein, and other substances having an active oxygen scavenging action; azulene, glycyrrhizin Acids and salts thereof, glycyrrhetinic acid and derivatives thereof, anti-inflammatory substances such as zinc oxide; vitamins and derivatives thereof such as riboflavin, tropherol, ascorbic acid, folic acid; jojoba oil, lanolin, liquid paraffin, squalane, isostearyl alcohol Oil etc. Ingredients: surfactants such as sodium stearyl sulfate, diethanolamine cesyl sulfate, glyceryl stearate; antioxidants such as sodium erythorbate; preservatives such as ethyl paraben; plant sterols; lipoproteins; bifidobacteria cultures, lactic acid bacteria cultures , Components such as extract of yeast, yeast extract, algal extract such as brown algae extract, red algae extract; blood circulation promoter such as γ-oryzanol; anti-seborrheic agent such as sulfur; fragrance; alcohol; Thickeners such as carboxy polymers; titanium yellow, safflower, and other colorants. When used in this way, it becomes a more general product, and thereby a synergistic effect with other active ingredients used in combination may lead to a superior effect than would normally be expected.

  The above-mentioned skin cosmetics suppress the growth of pigment cells and the production of melanin through the SCF expression increase inhibitory action of the SCF expression increase inhibitor as a constituent component, thereby preventing or improving spots, buckwheat, skin pigmentation, etc. And a whitening effect can be obtained.

  The SCF expression increase inhibitor of the present invention is suitably applied to humans, but can also be applied to animals other than humans as long as each effect is exhibited.

  Hereinafter, although a manufacture example, a test example, and a compounding example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all.

[Production Example 1] Manufacture of quince extract To 100 g of the crushed material of the coconut leaves, 1000 mL of 50% by mass ethanol (mass ratio of water to ethanol = 1: 1) was added and kept at 70 ° C. After gently stirring for 2 hours, the mixture was filtered using a filter paper to obtain an extract from the leaves of the quince. The obtained extract was concentrated under reduced pressure at 40 ° C., and then dried with a vacuum drier to obtain a pentagonal extract (sample 1). The yield of the obtained pentagonal extract was 28.7% by mass.

[Production Example 2] Production of licorice leaf extract 1000 g of 50% ethanol (mass ratio of water to ethanol = 1: 1) is added to 100 g of crushed licorice leaf, and the mixture is kept at 70 ° C. for 2 hours. After gently stirring, the mixture was filtered using a filter paper to obtain an extract from the leaves of licorice. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum dryer to obtain a licorice leaf part extract (sample 2). The yield of the obtained licorice leaf part extract was 21.7 mass%.

[Production Example 3] Production of Onion Strawberry Extract To 300 g of onion strawberries, add 2000 mL of 50 mass% ethanol (mass ratio of water to ethanol = 1: 1) and gently agitate for 3 hours while keeping at 80 ° C. Then, it filtered using the filter paper and obtained the extract from the root part of Oni strawberry. The obtained extract was concentrated under reduced pressure at 40 ° C., and then dried with a vacuum dryer to obtain an onion strawberry extract (sample 3). The yield of the onion strawberry extract obtained was 12.1% by mass.

[Production Example 4] Production of loquat extract To 100 g of loquat leaves, 2000 mL of 50 mass% ethanol (mass ratio of water and ethanol = 1: 1) was added and stirred gently for 3 hours while keeping at 70 ° C. Then, it was filtered using a filter paper to obtain an extract from loquat leaves. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum dryer to obtain a loquat extract (sample 4). The yield of the obtained loquat extract was 12.9% by mass.

[Production Example 5] Manufacture of Matylus extract To 200 g of crude crushed bark of Matylus, 2000 mL of 50 mass% ethanol (mass ratio of water to ethanol = 1: 1) was added and stirred gently for 2 hours while keeping at 80 ° C. After that, it was filtered using a filter paper to obtain an extract from the bark portion of Matylus. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum drier to obtain a Mathilus extract (sample 5). The yield of the resulting Matylus extract was 6.2% by mass.

[Production Example 6] Production of Melissa extract Add 50 ml of 50% ethanol (mass ratio of water to ethanol = 1: 1) to 50 g of Melissa whole plant and gently stir for 2 hours while keeping at 70 ° C. After that, it was filtered using a filter paper to obtain an extract from Melissa whole plant. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum dryer to obtain a Melissa extract (sample 6). The yield of the obtained Melissa extract was 30.7% by mass.

[Production Example 7] Manufacture of moon peach extract To 100 g of the crushed material of the moon peach leaf, 1000 mL of 50 mass% ethanol (mass ratio of water to ethanol = 1: 1) was added and kept at 70 ° C for 2 hours. After gently stirring, the mixture was filtered using a filter paper to obtain an extract from the leaves of moon peach. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum dryer to obtain a moon peach extract (sample 7). The yield of the obtained moon peach extract was 16.0 mass%.

[Production Example 8] Manufacture of pearl barley extract To 300 g of crushed seeds of pearl barley, 3000 mL of 50 mass% ethanol (mass ratio of water to ethanol = 1: 1) was added and gently stirred for 2 hours while keeping at 80 ° C. Then, the mixture was filtered using filter paper to obtain an extract from the seed part of pearl barley. The obtained extract was concentrated under reduced pressure at 40 ° C., and then dried with a vacuum dryer to obtain a pearl barley extract (sample 8). The yield of the obtained pearl barley extract was 2.0% by mass.

[Manufacturing Example 9] Manufacture of Senkyu Extract To 100 g of the crushed rhizome part (excluding root part), 500 mL of 90% by mass ethanol (mass ratio of water to ethanol = 1: 9) was added and kept at 80 ° C. Then, after gently stirring for 2 hours, the mixture was filtered using a filter paper to obtain an extract from the rhizome part (excluding the root part) of senkyu. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum drier to obtain a senkyu extract (sample 9). The yield of the resulting Senkyu extract was 10.0% by mass.

[Production Example 10] Production of tonin extract 500 g of 50 mass% ethanol (mass ratio of water to ethanol = 1: 1) was added to 100 g of crude tonin, and gently stirred for 2 hours while keeping at 80 ° C. Filtration was performed using filter paper to obtain an extract from tonin. The obtained extract was concentrated under reduced pressure at 40 ° C., and then dried with a vacuum dryer to obtain a tonin extract (sample 10). The yield of the obtained tonin extract was 13.3 mass%.

[Production Example 11] Production of gentian extract To 100 g of gentian roots and rhizomes, 2500 mL of ethanol was added and stirred gently for 2 hours while being kept at 80 ° C., followed by filtration using filter paper. An extract from the rhizome was obtained. The obtained extract was concentrated under reduced pressure at 40 ° C. and then dried with a vacuum dryer to obtain a gentian extract (sample 11). The yield of the obtained gentian extract was 14.8% by mass.

[Test Example 1] SCF mRNA expression increase inhibitory action test Each plant extract (samples 1 to 11) obtained in Production Examples 1 to 11 was tested for SCF mRNA expression increase suppressive action as follows.

(1) Preparation of single-stranded DNA Normal neonatal epidermal keratinocytes (NHEK) were seeded in a 35 mm dish and cultured under conditions of 37 ° C., 5% CO 2 -95% air for 24 hours. After completion of the culture, it was washed with 1 mL of a phosphate physiological buffer and replaced with 1 mL of Hanks' solution. Using an ultraviolet irradiation device equipped with four TOREX FL20SE-30 / DMRs as a light source, 50 mJ / cm ^{2 of} ultraviolet rays UV-B was irradiated. Immediately after the irradiation, the medium was replaced with a medium supplemented with a sample (samples 1 to 11) having a predetermined concentration (see Table 1) and cultured for 24 hours. After completion of the culture, total RNA was prepared by a conventional method. In addition, total RNA was similarly prepared for cells cultured with “no sample added / no UV irradiation” and “no sample added / with UV irradiation”. Total RNA was prepared using the following method.

  Dissolve the cells in 1 mL of RNA extraction reagent (product name: ISOGEN, Nippon Gene), add 200 μL of chloroform, isolate the upper RNA layer by centrifugation (12,000 rpm, 4 ° C., 15 minutes), and further isopropanol. Concentrated with. Concentrated and precipitated total RNA was dissolved in TE solution (10 mM Tris-HCl / 1 mM EDTA, pH = 8.0) to prepare a total RNA preparation, and a PCR device (product name: TaKaRa PCR Themal Cycler MP, manufactured by Takara Bio Inc.) ) And a real-time PCR dedicated reverse transcription kit (product name: TaKaRa ExScript RT reagent Kit, manufactured by Takara Bio Inc.), single-stranded DNA used as a template for measuring the expression level of SCF mRNA was synthesized.

(2) Real-time-PCR reaction using the cyber green method A sense primer and an antisense primer having the following sequences were prepared as primers for SCF gene amplification (manufactured by Takara Bio Inc.).
Sense primer: 5'-cccttaggaatgacagcagtagca-3 '
Antisense primer: 5'-gcccttgtaagacttggctgtctc-3 '

Moreover, the sense primer and antisense primer which have the following arrangement | sequence were produced as a G3PDH gene amplification primer as an internal standard (made by Takara Bio Inc.).
Sense primer: 5'-gcaccgtcaaggctgagaac-3 '
Antisense primer: 5'-atggtggtgaagacgccagt-3 '

  Single-stranded DNA prepared from total RNA preparations prepared from cells cultured in “No sample added / No UV irradiation”, “No sample added / With UV irradiation” and “Sample added / With UV irradiation”, respectively. Real-time PCR device (Product name: Real Time PCR System Smart Cycler II, manufactured by Cepheid) and Real-time PCR kit (Product name: SYBR Premix Ex Taq, manufactured by Takara Bio Inc.) A real-time PCR reaction was performed. In the single-stranded DNA solution for preparing a calibration curve, the relative value of the stock solution is set to “100,000” for convenience, and the concentration values “100,000”, “10000”, “1000”, “100” are repeatedly diluted 10 times thereafter. ”And“ 10 ”. The reaction was held at 95 ° C. for 10 seconds, followed by 45 cycles of 95 ° C. for 5 seconds and 57 ° C. for 20 seconds, and the amount of luminescence of the cyber green dye was measured for each cycle.

(3) Analysis An amplification curve of a DNA fragment encoding each of SCF and G3PDH was prepared from the amount of luminescence of the cyber green dye in each cycle. A calibration curve was prepared from the amplification curve of a dilution series of a single-stranded DNA solution for preparing a calibration curve, with the horizontal axis representing the concentration and the vertical axis representing the number of cycles that maximized the second derivative of the amplification curve. For each expression quantification sample, the number of cycles that maximized the second derivative of the amplification curve was plotted on a calibration curve, and the relative expression level was calculated. After correcting the expression level of SCF with the value of the expression level of G3PDH in the same sample, when the correction value of “no sample added / no UV irradiation” is set to 100, “no sample added / UV irradiation” and “ It was calculated as a correction value for “sample addition / with UV irradiation”.

The SCF mRNA expression increase rate (%) was calculated by the following formula.
mRNA expression increase suppression rate (%) = {(AB) − (AC)} / (AB) × 100
In the formula, A represents “correction value without sample addition / ultraviolet irradiation”, B represents “correction value without sample addition / ultraviolet irradiation”, and C represents “correction value with sample addition / ultraviolet irradiation”. Value ".
The results of the SCF mRNA expression increase inhibitory action test are shown in Table 1.

  As shown in Table 1, five coconut extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl extract, sensu extract, tonin extract It was confirmed that the product and the gentian extract have an excellent inhibitory effect on the increase in SCF mRNA expression.

[Formulation Example 1]
An emulsion having the following composition was produced by a conventional method.
Pentagram 50% by weight ethanol extract (Production Example 1) 0.01 g
Jojoba oil 4.0g
Placenta extract 0.1g
Olive oil 2.0g
Squalane 2.0g
Cetanol 2.0g
Glyceryl monostearate 2.0g
2.5 g of polyoxyethylene cetyl ether (20E.0)
Oleic acid polyoxyethylene sorbitan (20E.0) 2.0 g
Stearyl glycyrrhetinate 0.1g
1,3-butylene glycol 3.0 g
Hinokitiol 0.15g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

[Formulation Example 2]
A lotion having the following composition was produced by a conventional method.
Loquat leaf 50% by weight ethanol extract (Production Example 1) 2g
Glycerin 3g
1,3-butylene glycol 3g
Oleic acid polyoxyethylene sorbitan (20E.0) 0.5g
Methyl paraoxybenzoate 0.15g
Citric acid 0.1g
Sodium citrate 0.1g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

[Composition Example 3]
A cream having the following composition was produced by a conventional method.
50% by weight ethanol extract of Matylus bark (Production Example 5) 0.05g
Aloe extract 0.1g
Liquid paraffin 5.0g
Salami beeswax 4.0g
Squalane 10.0g
Cetanol 3.0g
Lanolin 2.0g
Stearic acid 1.0g
Oleic acid polyoxyethylene sorbitan (20E.0) 1.5g
3.0 g glyceryl monostearate
Oil soluble licorice extract 0.1g
1,3-butylene glycol 6.0 g
1.5 g of methyl paraoxybenzoate
Fragrance 0.1g
Purified water remainder (total amount is 100 g)

[Formulation Example 4]
A pack having the following composition was produced by a conventional method.
Melissa whole plant 50 mass% ethanol extract (Production Example 6) 0.05 g
Yokuinin extract 0.1g
Polyvinyl alcohol 15g
Polyethylene glycol 3g
7g of propylene glycol
Ethanol 10g
0.05 g of methyl paraoxybenzoate
0.1g dipotassium glycyrrhizinate
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

  The stem cell growth factor (SCF) expression increase inhibitor of the present invention is useful for the prevention or improvement of skin pigmentation, sun spots, buckwheat and the like after sunburn. The stem cell growth factor (SCF) expression increase inhibitor of the present invention is useful for the prevention or treatment of myelodysplastic syndrome, acute myeloid leukemia and the like.

Claims (2)

  Consists of five lion extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, sensu extract, tonin extract and gentian extract A stem cell growth factor (SCF) expression increase inhibitor comprising one or more plant extracts selected from the group as an active ingredient.   Consists of five lion extract, licorice leaf extract, oni strawberry extract, loquat extract, matylus extract, melissa extract, moon peach extract, pearl barley extract, sensu extract, tonin extract and gentian extract A prophylactic / therapeutic agent for diseases caused by increased expression of stem cell growth factor (SCF), comprising one or more plant extracts selected from the group as an active ingredient.