JP2012232920A - HEXOSAMINIDASE RELEASE INHIBITOR, STEM CELL GROWTH FACTOR mRNA EXPRESSION ELEVATION INHIBITOR, PROPHYLACTIC AND AMELIORATING AGENT OF HYDROGEN-PEROXIDE CYTOPATHY AND GLUTATHIONE PRODUCTION PROMOTOR - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a hexosaminidase release inhibitor, a stem cell growth factor mRNA expression elevation inhibitor, a prophylactic and ameliorating agent of hydrogen peroxide cytopathy, glutathione production promotor, elastase activity inhibitor, IV type collagen production promotor, hyaluronic acid production promotor, profilagrin production promotor, filagrin production promotor or involucrin production promotor, each from among highly safe natural products.SOLUTION: This hexosaminidase release inhibitor, stem cell growth factor mRNA expression elevation inhibitor, prophylactic and ameliorating agent of hydrogen peroxide cytopathy, glutathione production promotor, elastase activity inhibitor, IV type collagen production promotor, hyaluronic acid production promotor, profilagrin production promotor, filagrin production promotor or involucrin production promotor is incorporated with an extract from Piper longum as an effective component.

Description

  The present invention relates to a hexosaminidase release inhibitor, a stem cell growth factor mRNA expression increase inhibitor, a hydrogen peroxide cell disorder preventive / ameliorating agent, a glutathione production promoter, an elastase activity inhibitor, a type IV collagen production promoter, a hyaluron The present invention relates to an acid production promoter, a profilagrin production promoter, a filaggrin production promoter and an involucrin production promoter.

  Histamine release is a phenomenon in which histamine in mast cells is released extracellularly, and the released histamine causes an inflammatory reaction. Therefore, attempts have been made to prevent or treat allergic diseases and inflammatory diseases with substances that inhibit or suppress histamine release. However, it is difficult to directly evaluate the release of histamine, and the release of histamine can be evaluated using as an index the release of hexosaminidase that has been confirmed to be released simultaneously with the release of histamine. Therefore, by inhibiting the release of hexosaminidase, it is also possible to inhibit the release of histamine at the same time, thereby causing various skin diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other rough skin. It is considered effective for prevention, treatment or improvement of inflammatory diseases.

  It is also known that histamine mediates information transmission between cells as a local transmitter, enhances gastric acid secretion in the digestive tract, functions as a neurotransmitter in the central nervous system, and contributes to the maintenance of wakefulness. It has been. Here, excessive release of histamine causes ulcers due to excessive gastric acidity in the digestive tract, and contributes to sleep disorders in the central nervous system. As described above, by inhibiting the release of hexosaminidase, the release of histamine can also be inhibited at the same time, so that it is thought that this can prevent, treat or improve gastric ulcers, sleep disorders, etc. caused by excessive gastric acidity.

  As a plant extract having such a hexosaminidase release inhibitory effect, an extract from Fuji tea (see Patent Document 1) and the like are known.

  Stem cell factor (SCF), also called Mast 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. Stem cell growth factor is known to have an action of promoting proliferation and differentiation of pluripotent hematopoietic stem cells, germ cells, mast cells, megakaryocyte precursor cells, granulocyte / macrophage precursor cells, pigment cells, etc. . In addition, it is known that the expression of stem cell growth factor is enhanced by a spot site or ultraviolet irradiation (see Non-Patent Document 1).

  As the stem cell growth factor, a membrane-bound stem cell growth factor comprising 273 amino acid residues and a secretory stem cell growth factor that is cleaved by the action of proteolytic enzyme and released from the membrane are known. The membrane-bound stem cell growth factor binds to the stem cell growth factor receptor of the pigment cell while being bound to the keratinocytes and promotes the proliferation of the pigment cell. The secretory stem cell growth factor is cleaved at the binding site, released from the cell membrane, and bound to the stem cell growth factor receptor of the pigment cell, thereby promoting the proliferation of the pigment cell. Furthermore, stem cell growth factor is present in patients with acute myeloid leukemia in the presence of interleukin-3 (IL-3) and granulocyte macrophage colony stimulating factor (GM-CSF). It is known to promote the proliferation of myeloblasts (see Non-Patent Document 2).

  For this reason, abnormal production of stem cell growth factor leads to abnormal proliferation of pigment cells, which promotes melanin production and may cause spots, freckles, dullness, and the like. Abnormal production of stem cell growth factor 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 the expression of stem cell growth factor mRNA suppresses the proliferation of pigment cells, suppresses the excessive production of melanin in the skin, and is useful for the prevention or suppression of pigmentation, stains, freckles, etc. after sunburn. It is believed that there is. In addition, suppressing the increase in the expression of stem cell growth factor suppresses abnormal growth of myeloblasts and is considered useful for the prevention or treatment of myelodysplastic syndrome, acute myeloid leukemia and the like.

  Based on this idea, for example, extracts from Arnica (see Patent Document 2), amperopsin (see Patent Document 3) and the like are known as having an action of suppressing the production / release of stem cell growth factor. .

In recent years, active oxygen has attracted attention as a factor that oxidizes biological components, and adverse effects on living organisms have become a problem. Active oxygen is mainly generated in the process of energy metabolism in living cells, and is superoxide (that is, superoxide anion generated by one-electron reduction of oxygen molecules: .O ₂ ⁻ ), hydrogen peroxide (H ₂ O ₂ ). , Hydroxy radical (.OH) and singlet oxygen ( ¹ O ₂ ). These active oxygens are involved in the intracellular sterilization mechanism by phagocytic cells such as neutrophils and macrophages and play an important role in the removal of viruses and cancer cells, but excessive oxygen is generated Then, active oxygen may attack in vivo molecules constituting cell membranes and tissues and induce various diseases.

  For example, hydrogen peroxide is known to cause inflammation, production of lipid peroxides, denaturation / inactivation of various proteins, and DNA damage. Pulmonary diseases, cataracts, various arteriosclerosis (ischemic heart disease, myocardial infarction, cerebral ischemia, cerebral infarction, etc.), age-related aging, pigmentation of skin spots, neurodegenerative diseases ( Alzheimer's disease, Parkinson's disease, Huntington's chorea, etc.) and canceration are known.

  Therefore, if the cell damage caused by hydrogen peroxide can be prevented / improved and the homeostasis of the cells can be increased, the above-mentioned diseases caused by oxidative stress caused by hydrogen peroxide can be prevented / improved it is conceivable that. As an agent having such an action of preventing and improving hydrogen peroxide cell damage, an extract of a quince flower part (see Patent Document 4) and the like are known.

  Glutathione is a tripeptide composed of three amino acids, glutamic acid, cysteine and glycine, and is a compound having a major cysteine residue in the cell. Glutathione in cells functions as a radical donor, regulation of cell function by redox, foreign body metabolism, SH donor of various enzymes, and is also known as an antioxidant component. Its action expression is thought to be derived from cysteine residues. However, it has been reported that the amount of glutathione in cells is deficient or decreased due to excessive oxidative stress, addition of foreign substances, aging, etc., and this reduces the protective ability of cells against oxidative stress, and the cellular DNA In addition, it is considered to be a cause of damaging components such as proteins.

  In addition to the above-mentioned diseases that are induced by oxidative stress, diseases such as a decrease or deficiency in the amount of intracellular glutathione are known to be associated with the disease state. It is caused by drinking or ingestion of foreign substances such as heavy metals and chemical substances).

  That is, it is considered that promoting the production of glutathione can enhance the ability of cells to protect against oxidative stress, and can prevent and treat the above-mentioned disease group caused by the decrease or lack of intracellular glutathione level. . As those having such a glutathione production promoting action, a tenninka extract (see Patent Document 5), an extract of a gardenia plant (see Patent Document 6), and the like are known.

  The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as elastin, collagen and hyaluronic acid that are outside these cells and support the skin structure. In young skin, fibroblasts are actively proliferating, and the interaction between skin tissues such as fibroblasts and collagen keeps their homeostasis, ensuring moisture retention, flexibility, elasticity, etc. It is also kept fresh with tension and gloss.

  However, elastin, collagen, and hyaluron, which are the main components of the extracellular matrix, are affected by certain external factors such as UV irradiation, significant drying of air, excessive skin cleansing, and so on. As acid production decreases, it causes degradation and alteration. As a result, the moisturizing function and elasticity of the skin are lowered and abnormal exfoliation of the keratin occurs, so that the skin loses its tension and gloss, and exhibits aging symptoms such as rough skin and wrinkles. As described above, changes accompanying aging of the skin, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like are associated with reduction and degeneration of dermal matrix components such as elastin, collagen and hyaluronic acid. .

  Of the extracellular matrix components described above, elastin is a fiber that gives elasticity to the skin tissue. When elastin is decomposed with aging or the like, the elasticity of the skin is lost and the elasticity is lowered. Elastase, which is an enzyme that degrades elastin, is activated by irradiation with ultraviolet rays, thereby accelerating the degradation of elastin. Therefore, it is considered that by inhibiting the activity of elastase, the degradation of elastin can be suppressed, and skin aging symptoms such as loss of tension and reduced elasticity can be prevented and improved.

  In addition, it is known that elastase activity in vivo is enhanced by smoking or the like. Since the lung substrate is composed of collagen and elastin, it is considered that if the elastase activity is increased by smoking or the like, the alveolar wall may be destroyed to cause emphysema and the like. Furthermore, it is considered that the lung capillaries are destroyed by the increase in the activity of elastase, which may lead to acute respiratory distress syndrome (ARDS) such as pulmonary edema. Therefore, if the activity of elastase in vivo can be inhibited, it is considered that respiratory diseases such as emphysema and pulmonary edema can be prevented and treated.

  Conventionally, a star fruit fruit extract etc. are known as what has an elastase activity inhibitory effect (refer patent document 7).

  On the other hand, among the extracellular matrix components described above, type IV collagen is a main component of the basement membrane present at the boundary between the epidermis and dermis. The functions of the basement membrane are diverse, involved in the adhesion of the epidermis to the dermis, the determination of the polarity of the epidermis, the control of epidermal differentiation and proliferation, and the regulation of nutrients derived from factors and blood components produced by the dermal cells Yes. Therefore, the basement membrane plays an extremely important role in maintaining the structure and homeostasis of the skin.

  However, the basement membrane changes in structure or decreases in function due to aging, and thus exhibits skin aging symptoms such as wrinkles and sagging. In particular, if the production of type IV collagen, the main component of the basement membrane, decreases, the structure of the basement membrane changes and skin aging symptoms such as wrinkles and sagging appear. By doing so, it is considered that these skin aging symptoms and the like can be prevented and improved.

  Known examples of such a type IV collagen production promoting action include phlorizin and phloretin (see Patent Document 8), an extract from black-spotted azuki bean (see Patent Document 9), and the like.

  On the other hand, among the above-mentioned extracellular matrix components, hyaluronic acid is a kind of mucopolysaccharide, and when hyaluronic acid is reduced or denatured with aging or irradiation with ultraviolet rays, skin roughness, wrinkles, dullness, and disappearance of texture are lost. In addition, aging symptoms such as a decrease in elasticity and a decrease in moisture retention function are exhibited. Therefore, it is considered that aging symptoms of skin can be prevented and improved by promoting the production of hyaluronic acid.

  In addition to skin tissue, hyaluronic acid is present in cartilage, joint fluid, umbilical cord, ophthalmic vitreous, and other connective tissues. Hyaluronic acid has a function of retaining cells by filling the gaps between cells, and also retains moisture in the cell gaps, imparts lubricity and flexibility to tissues, and resists external forces such as mechanical damage. It has many functions such as resistance.

  Among these, the hyaluronic acid contained in the joint fluid covers the surface of the articular cartilage and is useful for the smooth operation of the joint due to the lubricating function of the hyaluronic acid, the cartilage covering / protecting function, and the like. On the other hand, in arthritis such as rheumatoid arthritis, it is known that the concentration of hyaluronic acid in joint fluid is reduced. Therefore, it is considered that by promoting the production of hyaluronic acid, arthritis such as rheumatoid arthritis, osteoarthritis, septic arthritis, gouty arthritis, traumatic arthritis, or osteoarthritis can be prevented or treated.

  Furthermore, in the healing process of wounds or burns, granulation (tissue) is formed, but it is known that hyaluronic acid significantly increases in the granulation. Therefore, it is considered that healing of wounds or burns can be promoted by promoting production of hyaluronic acid.

  Conventionally known extracts having hyaluronic acid production promoting action include an extract from Kusunohagashi (see Patent Document 10), an extract from the fruit of a star fruit (see Patent Document 11), and the like.

  Filaggrin is a structural component of the skin, is involved in the barrier function in the skin, and is considered to have a function of preventing the entry of allergens, toxins and infectious organisms. Reduced function due to filaggrin gene mutation is associated with the risk of developing atopic diseases including atopic dermatitis (eczema, skin inflammation, skin itch, etc.), allergy, asthma, etc. In some cases, it is known to lead to skin diseases such as ichthyosis vulgaris (see Non-Patent Document 3).

  On the other hand, amino acids which are the main components of natural moisturizing factors (NMF) are produced by the degradation of filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilagrin in epidermal keratinocytes present in the granule layer immediately below the stratum corneum. It is then phosphorylated immediately, accumulated in keratohyalin granules, decomposed to filaggrin through dephosphorylation and hydrolysis, transferred to the stratum corneum, increased the efficiency of keratin filament aggregation, and is involved in the internal construction of keratinocytes It is known (see Non-Patent Document 4). In recent years, it has been known that this filaggrin is very important and indispensable for moisture retention of the skin, and that the synthetic power of filaggrin is reduced by conditions such as drying, and the amount of amino acids in the stratum corneum is reduced ( Non-patent document 5).

  Therefore, by promoting filaggrin production in epidermal keratinocytes, it is possible to prevent, treat or ameliorate atopic diseases including atopic dermatitis (eczema, skin inflammation, skin itch, etc.), allergies, asthma, etc. Conceivable. In addition, it is expected that the water environment of the stratum corneum can be essentially improved by promoting the production of filaggrin and thereby increasing the amount of amino acids in the stratum corneum.

  Examples of such profilagrin production promoter or filaggrin production promoter include licorice extract (see Patent Document 12), liquiritin known as flavanone glycoside contained in natural plants (see Patent Document 13), Citrus, and the like. Plant extracts or yeast extracts belonging to the genus (see Patent Document 14) and the like are known.

  Corneocytes are composed of keratinized fibers (cornified envelope, CE) containing keratin fibers as the main component and enveloping them. This CE is formed by cross-linking and insolubilizing a plurality of CE precursor proteins produced in accordance with the differentiation of epidermal keratinocytes by the enzyme transglutaminase. Furthermore, ceramide and the like are covalently bonded to form a hydrophobic structure, whereby a foundation of a lamellar structure of intercellular lipid is supplied to a part of CE, thereby forming the basis of the stratum corneum barrier function.

  As one of the CE precursor proteins, involucrin is known. By promoting the production of such involucrin, the stratum corneum barrier function is improved, skin aging symptoms such as rough skin and dry skin, atopic dermatitis It is thought that dry skin diseases such as psoriasis, ichthyosis and psoriasis can be prevented, treated or ameliorated. Based on such an idea, conventionally, a seed extract of Ceylon tetsuboku has been known as having an involucrin production promoting action (see Patent Document 15).

JP 2003-12532 A JP 2008-169118 A JP 2009-209050 A JP 2006-8571 A JP 2008-285422 A JP 2006-347934 A Japanese Patent Laid-Open No. 2003-300893 JP 2007-106712 A JP 2007-217352 A JP 2003-146837 A Japanese Patent Laid-Open No. 2003-300893 JP 2002-363054 A JP 2003-146886 A JP 2001-261568 A JP 2005-213187 A

"J. Invest. Dermatol.", 2001, Vol.116, Issue 4, p.578-586 "Blood", 1992, Vol.80, No.1, p.60-67 "Nat Genet.", 2006, Vol.38, No.4, p.441-446 "Fragrance Journal Extra Edition", 2000, Vol.17, p.14-19 "Arch. Dermatol. Res.", 1996, Vol.288, p.442-446

  The present invention is an excellent hexosaminidase release inhibitory action, stem cell growth factor mRNA expression increase inhibitory action, hydrogen peroxide cell damage prevention / amelioration action, glutathione production promoting action, elastase activity from among highly safe natural products Hexosaminidase release inhibitor containing an inhibitory action, type IV collagen production promoting action, hyaluronic acid production promoting action, profilagulin production promoting action, filaggrin production promoting action or involucrin production promoting action , Stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluronic acid production promoter, profilagrin production promoter, Filaggrin production promoter or involucrin production promoter It aims to provide.

  In order to solve the above-mentioned problems, the hexosaminidase release inhibitor, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, IV Type collagen production promoter, hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter is characterized by containing an extract from hihatsu as an active ingredient.

  According to the present invention, a hexosaminidase release inhibitor having an excellent hexosaminidase release inhibitory action and a high safety, a stem cell growth factor mRNA expression rise inhibitory action having an excellent inhibitory action on stem cell growth factor mRNA expression and high safety. Agent, hydrogen peroxide cell disorder preventive / ameliorating action that is excellent and safe, hydrogen peroxide cell disorder preventing / ameliorating action, glutathione production promoting action that is safe and highly safe, glutathione production promoter, elastase activity inhibitory action Excellent and safe elastase activity inhibitor, excellent type IV collagen production promoting action and highly safe type IV collagen production promoting agent, hyaluronic acid production promoting action and safe hyaluronic acid production promoting agent, Profilagrin production promoter, filaggrin production, which has excellent profilaggrin production promoting action and high safety It can be provided to promote good and high safety filaggrin production accelerator to the action, or superior to involucrin production promoting action and high safety involucrin production accelerator.

Embodiments of the present invention will be described below.
Hexosaminidase release inhibitor, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder prevention / amelioration agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluron The acid production promoter, profilagrin production promoter, filaggrin production promoter, or involucrin production promoter contains an extract from hihitsu as an active ingredient.

  Here, in the present embodiment, the “extract from baboon” includes an extract obtained from baboon as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or Any of these crudely purified products or purified products is included.

The extraction raw material used in this embodiment is Hihatsu (scientific name: Piper longum Linn.).
Gibbon (Piper longum Linn.) Is an evergreen climbing plant belonging to the genus Pepperaceae, and is widely distributed in Southeast Asia and can be easily obtained from these regions. In addition, Hibachi's fruit spikes are fleshy and thick cylindrical, and are used as spices.

  Examples of the constituent parts of the rabbit that can be used as the extraction raw material include fruit ears, roots, leaves, stems, flowers, etc., and one or more of these can be used as the extraction raw material. It is particularly preferable to use fruit spikes.

  The extract from Hihatsu can be obtained by drying the raw material for extraction and then pulverizing the raw material as it is or using a crusher and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or 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 a pretreatment such as degreasing, extraction with a polar solvent of hihatsu can be performed efficiently.

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

  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 this embodiment includes purified water, hot water, ion exchange 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 mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by volume of a lower aliphatic alcohol with respect to 10 parts by volume of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by volume of a lower aliphatic ketone with 10 parts by volume of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by volume of a polyhydric alcohol with respect to the volume 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 an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  The extract obtained as described above can be used as it is, a hexosaminidase release inhibitor, a stem cell growth factor mRNA expression increase inhibitor, a hydrogen peroxide cell disorder prevention / amelioration agent, a glutathione production promoter, an elastase activity. It can be used as an active ingredient of inhibitors, type IV collagen production promoters, hyaluronic acid production promoters, profilagrin production promoters, filaggrin production promoters or involucrin production promoters, It is easier to use.

  In addition, since the extract from Hihatsu has a peculiar odor, it can be purified for the purpose of decoloring, deodorizing, etc. within a range that does not cause a decrease in its physiological activity. Since it is not used in a large amount when it is blended in, there is no practical problem even if it is not purified.

  The extract from baboon obtained as described above has excellent hexosaminidase release inhibitory action, stem cell growth factor mRNA expression increase inhibitory action, hydrogen peroxide cell damage preventive / ameliorating action, glutathione production promoting action, elastase Activity inhibitory action, type IV collagen production promoting action, hyaluronic acid production promoting action, profilagulin production promoting action, filaggrin production promoting action and involucrin production promoting action, hexosaminidase release inhibitor, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter And as an active ingredient of involucrin production promoter It is possible to have.

  Hexosaminidase release inhibitor of this embodiment, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluron The acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter may be composed only of an extract from baboon, or may be formulated from an extract from baboon.

  Hexosaminidase release inhibitor of this embodiment, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluron The acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter is powdered in a conventional manner using a pharmaceutically acceptable carrier such as dextrin or cyclodextrin or any other auxiliary agent. It can be formulated into any dosage form such as granules, tablets and liquids. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. Hexosaminidase release inhibitor, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder prevention / amelioration agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluronic acid production promoter , Profilagrin production promoter, filaggrin production promoter or involucrin production promoter can be used in combination with other compositions (for example, topical skin preparations, cosmetic foods and drinks), ointments, external use It can be used as a liquid, a patch or the like.

  Hexosaminidase release inhibitor of this embodiment, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluron When formulating an acid production promoter, profilaggrin production promoter, filaggrin production promoter, or involucrin production promoter, the content of the extract from baboon is not particularly limited and is appropriately set according to the purpose. can do.

  In addition, the hexosaminidase release inhibitor, the stem cell growth factor mRNA expression increase inhibitor, the hydrogen peroxide cell disorder preventive / improving agent, the glutathione production promoter, the elastase activity inhibitor, the type IV collagen production promoter of the present embodiment , Hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter, if necessary, hexosaminidase release inhibitory action, stem cell growth factor mRNA expression increase inhibitory action, hydrogen peroxide cells Other naturals that have the effect of preventing or improving disorders, glutathione production promoting action, elastase activity inhibiting action, type IV collagen production promoting action, hyaluronic acid production promoting action, profilaggrin production promoting action, filaggrin production promoting action or involucrin production promoting action Extracts, etc., with extracts from Hihatsu It can be used as an active ingredient by blending the.

  Hexosaminidase release inhibitor of this embodiment, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluron Examples of the method of administering an acid production promoter, profilagrin production promoter, filaggrin production promoter or involucrin production promoter to a patient include transdermal administration, oral administration, etc., depending on the type of the disease, A method suitable for treatment or the like may be selected as appropriate.

  Further, the hexosaminidase release inhibitor, the stem cell growth factor mRNA expression increase inhibitor, the hydrogen peroxide cell disorder preventive / improving agent, the glutathione production promoter, the elastase activity inhibitor, the type IV collagen production promoter of the present embodiment The dosage of hyaluronic acid production promoter, profilagrin production promoter, filaggrin production promoter or involucrin production promoter should also be increased or decreased appropriately depending on the type of disease, severity, individual differences of patients, administration method, administration period, etc. Good.

  The hexosaminidase release inhibitor of the present embodiment is a contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, etc., through the hexosaminidase release inhibitory action of the extract from Hihatsu. Various skin diseases associated with rough skin, rheumatoid arthritis, osteoarthritis, asthma and the like can be prevented, treated or improved. Further, the hexosaminidase release inhibitor of the present embodiment prevents, treats or improves gastric ulcers, sleep disorders, etc. caused by excessive gastric acid through a hexosaminidase release inhibitory action of an extract from Japanese cherries. Can do. However, the hexosaminidase release inhibitor of the present embodiment can be used for all uses other than these uses that are meaningful for exerting the hexosaminidase release inhibitory action.

  The stem cell growth factor mRNA expression increase inhibitor of the present embodiment can suppress the increase in expression of stem cell growth factor through the action of suppressing the increase in stem cell growth factor mRNA expression possessed by the extract from Japanese red pepper. 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. In addition, the stem cell growth factor mRNA expression increase inhibitor of the present embodiment can suppress the abnormal growth of myeloblasts through the suppressive action on the increase in stem cell growth factor mRNA expression of the extract from chickpeas, and thereby, Diseases such as formation syndrome and acute myeloid leukemia can be prevented, treated or ameliorated. However, the stem cell growth factor mRNA expression increase suppressant of the present embodiment can be used for all purposes that are meaningful for exhibiting the stem cell growth factor mRNA expression increase suppression effect in addition to these uses.

  The agent for preventing / ameliorating hydrogen peroxide cell damage according to the present embodiment is that the excessive production of hydrogen peroxide is associated with the pathological condition through the action of preventing / ameliorating hydrogen peroxide cell damage of the extract from Hihatsu. It can be used for treatment / prevention of known diseases. Such diseases include pulmonary diseases caused by smoking, cataracts, various arteriosclerosis (ischemic heart disease, myocardial infarction, cerebral ischemia, cerebral infarction, etc.), age-related aging phenomenon, skin spots, etc. Examples include pigmentation, neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, Huntington's chorea, etc.), and canceration. However, the hydrogen peroxide cell disorder preventive / ameliorating agent of the present embodiment can be used for all purposes that are meaningful for exerting the hydrogen peroxide cell disorder preventive / ameliorating action in addition to these uses. it can.

  The glutathione production promoter according to the present embodiment prevents, treats, or improves a disease or the like in which a decrease or deficiency in intracellular glutathione is known to be associated with a pathological condition through a glutathione production promoting action of an extract from Hihatsu. can do. Such diseases include pulmonary diseases caused by smoking, cataracts, various arteriosclerosis (ischemic heart disease, myocardial infarction, cerebral ischemia, cerebral infarction, etc.), age-related aging phenomenon, skin spots, etc. Pigmentation, neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, Huntington's chorea, etc.), canceration, liver damage (caused by excessive drinking of alcohol or ingestion of foreign substances such as heavy metals and chemical substances), and the like. However, the glutathione production promoter of the present embodiment can be used for all purposes other than these uses that are meaningful for exerting the glutathione production promoting action.

  The elastase activity inhibitor of the present embodiment suppresses elastin degradation through the elastase activity inhibitory action of the extract from baboon, and prevents, treats or improves aging symptoms such as disappearance of skin tension and decreased elasticity. can do. Moreover, the elastase activity inhibitor of this embodiment can be used as an active ingredient of the pharmaceutical for prevention and treatment of the disease resulting from the elastase activity enhancement, or a quasi-drug. Examples of such diseases include respiratory diseases such as emphysema and pulmonary edema. However, the elastase activity inhibitor of the present embodiment can be used for all uses other than these uses, which are meaningful for exerting an elastase activity inhibitory action.

  The type IV collagen production promoter of the present embodiment can promote type IV collagen production in the epidermis and promote remodeling of the epidermis basement membrane through the type IV collagen production promotion effect of the extract from Hihatsu. Thus, aging of the skin such as wrinkles and sagging can be prevented or improved. However, the type IV collagen production promoter of the present embodiment can be used for all purposes that are meaningful in exerting the type IV collagen production promotion effect in addition to these uses.

  The hyaluronic acid production promoter of the present embodiment promotes hyaluronic acid production through the hyaluronic acid production promoting action of the extract from hihatsu, roughening, wrinkles, dullness, disappearance of texture, reduced elasticity and moisturizing function It is possible to prevent or ameliorate skin aging symptoms such as lowering of the skin. Further, the hyaluronic acid production promoter of the present embodiment can prevent or treat arthritis such as rheumatoid arthritis, osteoarthritis, septic arthritis, gouty arthritis, traumatic arthritis, or osteoarthritis, and wounds. Or it can promote the healing of burns. However, the hyaluronic acid production promoter of the present embodiment can be used for all purposes that are meaningful for exerting hyaluronic acid production promoting action in addition to these uses.

  The profilaggrin production promoter of this embodiment promotes the production of profilaggrin in cells through the profilaggrin production promoting action possessed by the extract from Hihatsu, and increases the amount of filaggrin obtained by hydrolysis of profilagrin. The skin moisturizing ability can be improved, thereby maintaining the elasticity of the skin and preventing, treating or improving skin aging, rough skin and the like. In addition, the profilaggrin production promoter of the present embodiment promotes the production of profilagrin in the cell and increases the amount of filaggrin through the profilaggrin production promoting action of the extract from Japanese chickweed, and thereby the skin barrier function. Atopic dermatitis (eczema, skin inflammation, skin itch, etc.), allergy, asthma and the like can be prevented, treated or improved. However, the profilaggrin production promoter of this embodiment can be used for all the uses meaningful in exhibiting the profilaggrin production promotion effect besides these uses.

  The filaggrin production promoter of the present embodiment can promote the production of filaggrin in cells through the filaggrin production promoting action of the extract from chickweed, and can improve the skin moisturizing ability. The elasticity can be maintained, and skin aging, rough skin, etc. can be prevented, treated or improved. Moreover, the filaggrin production promoter of this embodiment promotes the production of filaggrin in the cell through the filaggrin production promoting action of the extract from Hihatsu and enhances the barrier function of the skin, thereby atopic dermatitis ( Eczema, skin inflammation, skin itch, etc.), allergies, asthma and the like can be prevented, treated or ameliorated. However, the filaggrin production promoter of the present embodiment can be used for all purposes that are meaningful for exerting a filaggrin production promoting action in addition to these uses.

  The involucrin production promoter of the present embodiment, through the involucrin production promoting action of the extract from baboon, aging symptoms such as rough skin, dry skin, ichthyosis, psoriasis, atopic dermatitis, psoriasis, etc. It can prevent, treat or ameliorate dry skin diseases and the like. However, the involucrin production promoter of the present embodiment can be used for all purposes that are meaningful for exerting the involucrin production promoting action in addition to these uses.

  Further, the hexosaminidase release inhibitor, the stem cell growth factor mRNA expression increase inhibitor, the hydrogen peroxide cell disorder preventive / improving agent, the glutathione production promoter, the elastase activity inhibitor, the type IV collagen production promoter of the present embodiment , Hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter is an excellent hexosaminidase release inhibitory effect, stem cell growth factor mRNA expression increase inhibitory effect, prevention of hydrogen peroxide cell damage・ Since it has an improving action, glutathione production promoting action, elastase activity inhibiting action, type IV collagen production promoting action, hyaluronic acid production promoting action, profilagrin production promoting action, filaggrin production promoting action or involucrin production promoting action, for example, for external use on the skin Good for blending in preparations or foods It is. In this case, an extract from baboon may be added as it is, or a hexosaminidase release inhibitor, a stem cell growth factor mRNA expression increase inhibitor formulated from an extract from baboon, hydrogen peroxide cell damage A prophylactic / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter may be added.

  Here, the topical skin preparation is not limited in its category, and includes a wide range of skin cosmetics, quasi-drugs, pharmaceuticals, and the like used transdermally. Creams, milky lotions, beauty essences, lotions, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  The food and drink is not limited in its category, and includes a wide variety of foods such as general foods, health foods, health functional foods, quasi drugs, and pharmaceuticals that are taken orally.

  Further, the hexosaminidase release inhibitor, the stem cell growth factor mRNA expression increase inhibitor, the hydrogen peroxide cell disorder preventive / improving agent, the glutathione production promoter, the elastase activity inhibitor, the type IV collagen production promoter of the present embodiment , Hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter is an excellent hexosaminidase release inhibitory effect, stem cell growth factor mRNA expression increase inhibitory effect, prevention of hydrogen peroxide cell damage・ Histamine release mechanism because it has an improving action, glutathione production promoting action, elastase activity inhibiting action, type IV collagen production promoting action, hyaluronic acid production promoting action, profilagline production promoting action, filaggrin production promoting action or involucrin production promoting action , Stem cell growth factor expression machine , Molecular mechanism of cell damage caused by hydrogen peroxide, glutathione production mechanism, elastase activity control mechanism, type IV collagen production mechanism, hyaluronic acid production mechanism, profilaggrin / filaggrin production mechanism or involucrin production mechanism It can be suitably used as a reagent for research.

  In addition, the hexosaminidase release inhibitor, the stem cell growth factor mRNA expression increase inhibitor, the hydrogen peroxide cell disorder preventive / improving agent, the glutathione production promoter, the elastase activity inhibitor, the type IV collagen production promoter of the present embodiment , Hyaluronic acid production promoter, profilaggrin production promoter, filaggrin production promoter or involucrin production promoter is preferably applied to humans, but as long as each effect is exerted, other than human The present invention can also be applied to other animals (eg, mouse, rat, hamster, dog, cat, cow, pig, monkey, etc.).

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In this test example, a freeze-dried product of Hihatsu extract BG (manufactured by Maruzen Pharmaceutical, sample 1) was used as an extract from Hihatsu.

[Test Example 1] Hexosaminidase release inhibitory action test The above-mentioned Hihatsu extract (sample 1) was tested for hexosaminidase release inhibitory action as follows.

Rat basophil leukemia cells (RBL-2H3) were cultured in S-MEM medium supplemented with 15% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with S-MEM medium supplemented with 15% FBS to a cell density of 4.0 × 10 ⁵ cells / mL, and DNP-specific IgE was added to a final concentration of 0.5 μL / mL. Thereafter, 100 μL per well was seeded in a 96-well plate and cultured overnight.

  After completion of the culture, the medium was removed, and washing was performed twice with 100 μL of Silligalian buffer. Next, 10 μL of the same buffer solution and 30 μL of the same buffer solution to which the test sample (sample 1, sample concentration is shown in Table 1 below) was added, or 40 μL of the same buffer solution without addition of the sample were added to each well, and the temperature was changed to 37 ° C. Left for 10 minutes. Thereafter, 10 μL of a 100 ng / mL DNP-BSA solution was added, and the mixture was allowed to stand at 37 ° C. for 15 minutes to release hexosaminidase.

  Thereafter, the release was stopped by allowing the 96-well plate to stand on ice. 10 μL of the cell supernatant of each well was collected in a new 96-well plate, 10 μL of 1 mM p-NAG (p-nitrophenyl-N-acetyl-β-D-glucosaminide) solution was added to each well, and the mixture was incubated at 37 ° C. The reaction was carried out for 1 hour.

After completion of the reaction, 250 μL of 0.1 M Na ₂ CO ₃ / NaHCO ₃ was added to each well, the absorbance at wavelengths 415 nm and 650 nm was measured, and the value obtained by subtracting the absorbance at 650 nm from the absorbance at 415 nm was used as the correction value. In addition, as a blank test, absorbance at wavelengths of 415 nm and 650 nm of a mixed solution of 10 μL of cell supernatant and 250 μL of 0.1 M Na ₂ CO ₃ / NaHCO ₃ was measured, and a correction value was calculated. From the obtained measurement results, the hexosaminidase release inhibition rate (%) was calculated by the following formula.

Inhibition rate of hexosaminidase release (%) = {1− (B−C) / A} × 100
In the formula, A represents “correction value without addition of sample”, B represents “correction value with addition of test sample”, and C represents “correction value with addition of test sample / no p-NAG”. .
The results are shown in Table 1.

  As shown in Table 1, it was confirmed that the Hihatsu extract (Sample 1) has an excellent hexosaminidase release inhibitory action.

[Test Example 2] Stem cell growth factor (SCF) mRNA expression increase inhibitory action test The above-mentioned Hihatsu extract (Sample 1) was tested for SCF mRNA expression increase suppressive action as follows.

Normal human epidermal keratinocyte (NHEK) in an 80 cm ² flask using a growth medium (EpiLife-KG2) for long-term culture of human normal epidermal keratinocytes at 37 ° C., 5% CO ₂ -95% air The cells were precultured under the above conditions and cells were collected by trypsin treatment. The collected cells are diluted with Epilife-KG2 medium so as to have a cell density of 20 × 10 ⁴ cells / mL, and then seeded in 2 mL each in a 35 mm petri dish (manufactured by FALCON) (40 × 10 ⁴ cells / petri dish). ° _C., and cultured for 24 hours under the conditions of 5% CO 2 -95% air.

After culture, the medium was removed, 1 mL of HEPES buffer was added, UV-B irradiation (50 mJ / cm ² ) was performed, and then the test life (sample 1, sample concentration: see Table 2 below) was added to Epilife-KG2 medium Alternatively, 2 mL of Epilife-KG2 medium without any sample was added to each dish, and cultured for 24 hours under conditions of 37 ° C. and 5% CO ₂ -95% air. After culturing, the medium is removed, and total RNA is extracted with ISOGEN (Nippon Gene, Cat. No. 311-02501), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, the expression level of mRNA of SCF and GAPDH as an internal standard was measured. The detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR PrimeScript RT-PCR Kit (Perfect Real Time, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). The expression level of SCF mRNA is corrected by the value of GAPDH mRNA based on the total RNA preparation prepared from cells cultured without UV irradiation / sample addition, UV irradiation / sample addition and UV irradiation / sample addition, respectively. Further, the correction values of ultraviolet irradiation / no sample addition and ultraviolet irradiation / sample addition were calculated when the correction value of UV non-irradiation / no sample addition was 100. From the obtained results, the SCF mRNA expression increase suppression rate (%) was calculated by the following formula.

SCF mRNA expression increase suppression rate (%) = {(A−B) − (A−C)} / (A−B) × 100
In the formula, A represents “correction value when no UV irradiation is performed and no sample is added”, B represents “correction value when UV irradiation is performed and no sample is added”, and C is “correction value when UV irradiation is performed and sample is not added”. Represents.
The results are shown in Table 2.

  As shown in Table 2, it was confirmed that the Hihatsu extract (Sample 1) has an excellent inhibitory effect on the increase in SCF mRNA expression.

[Test Example 3] Cytotoxicity-inhibiting action test against hydrogen peroxide The above-mentioned extract of baboon (sample 1) was tested for its cytotoxicity-inhibiting action against hydrogen peroxide by the following test method.

Human normal skin fibroblasts (NB1RGB) were cultured using α-MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with 5% FBS-containing α-MEM medium to a cell density of 2.5 × 10 ⁵ cells / mL, then seeded at 200 μL per well in a 48-well plate, and cultured overnight.

  After culturing, the medium was removed, and 1% FBS-containing α-MEM medium to which a test sample (sample 1, sample concentration is shown in Table 3 below) was added or 1% FBS-containing α-MEM medium to which no sample was added was added to each well. 200 μL was added to each and cultured for 24 hours. After completion of the culture, the medium is removed from each well, washed with 400 μL of PBS buffer, Hank's buffer in which hydrogen peroxide is dissolved (hydrogen peroxide final concentration: 1 mM), or Hank 'without hydrogen peroxide. 200 μL of s buffer was added to each well and incubated for 2 hours.

  After culturing, the plate was washed with 400 μL of PBS buffer, 200 μL of neutral red solution dissolved in 1% FBS-containing α-MEM medium at a final concentration of 0.05 mg / mL was added and cultured for 2.5 hours. After incubation, the neutral red solution was removed, and 300 μL of ethanol / acetic acid solution (ethanol: acetic acid: water = 50: 1: 49) was added to each well to extract the dye. The absorbance at 540 nm of the obtained extract was measured. From the measurement results, the hydrogen peroxide damage inhibition rate (%) was calculated by the following formula.

Hydrogen peroxide damage inhibition rate (%) = {1- (C−A)} / (C−B)} × 100
In the above formula, A represents “absorbance at the time of test sample addition / hydrogen peroxide treatment”, B represents “absorbance at the time of no sample addition / hydrogen peroxide treatment”, and C represents “absorbance at no sample addition / hydrogen peroxide treatment”. It represents “absorbance without treatment”.
The results of the above test are shown in Table 3.

  As shown in Table 3, it was confirmed that the Hihatsu extract (Sample 1) has an excellent hydrogen peroxide disorder inhibiting action.

[Test Example 4] Glutathione production promoting test-1
With respect to the above-mentioned extract of baboon (sample 1), the glutathione production promoting effect on B16 melanoma cells was tested as follows.

B16 melanoma cells were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS to a cell density of 10 × 10 ⁴ cells / mL, and then seeded at 200 μL per well in a 48-well plate and cultured overnight.

  After the culture, the medium was removed, and 200 μL of 10% FBS-containing Dulbecco MEM medium to which the test sample (sample 1, sample concentration is shown in Table 4) or 10% FBS-containing Dulbecco MEM medium without the sample added was added to each well. Added and incubated for 24 hours. After completion of the culture, the medium was removed from each well, washed with 400 μL of PBS buffer, and cells were lysed using 150 μL of M-PER (manufactured by PIERCE).

  Of these, 100 μL was used to quantify total glutathione. That is, after adding 100 μL of cell extract dissolved in 96-well plate, 50 μL of 0.1 M phosphate buffer, 25 μL of 2 mM NADPH and 25 μL of glutathione reductase (final concentration 17.5 units / mL), the mixture was heated at 37 ° C. for 10 minutes. 25 μL of 10 mM 5,5′-dithiobis (2-nitrobenzoic acid) was added, and the absorbance at a wavelength of 412 nm until 5 minutes was measured to determine ΔOD / min. The total glutathione concentration was calculated based on a calibration curve prepared using oxidized glutathione (manufactured by Wako Pure Chemical Industries, Ltd.). After correcting the obtained value to the amount of glutathione per total protein amount, the glutathione production promotion rate (%) was calculated by the following formula.

Glutathione production promotion rate (%) = B / A × 100
In the formula, A represents “the amount of glutathione per total protein in the cell when no sample is added (control)”, and B represents “the amount of glutathione per total amount of protein in the cell when added to the test sample”.
The results are shown in Table 4.

  As shown in Table 4, it was confirmed that the Hihatsu extract (Sample 1) has an excellent glutathione production promoting action on B16 melanoma cells.

[Test Example 5] Glutathione production promoting action test-2
With respect to the above extract of the cherries (Sample 1), the glutathione production promoting effect on epidermal keratinocytes was tested as follows.

Human normal neonatal epidermal keratinocytes (NHEK) were cultured using a growth medium for long-term culture of human normal epidermal keratinocytes (EpiLife-KG2), and then cells were collected by trypsin treatment. The collected cells were diluted with EpiLife-KG2 medium to a cell density of 1.0 × 10 ⁵ cells / mL, then seeded at 500 μL per well in a collagen-coated 24-well plate and cultured overnight.

  After culturing, the medium is removed, and 500 μL of EpiLife-KG2 medium to which the test sample (sample 1, sample concentration is shown below) is added or EpiLife-KG2 medium to which no sample is added is added to each well, and further 24 hours Cultured. After completion of the culture, the medium was removed from each well, washed with 1 mL of PBS buffer, and cells were lysed using 150 μL of M-PER (PIERCE).

Of these, 100 μL was used, and the total glutathione was quantified in the same manner as in Test Example 4 described above, and the glutathione production promotion rate (%) was calculated.
The results are shown in Table 5.

  As shown in Table 5, it was confirmed that the Hihatsu extract (Sample 1) has an excellent glutathione production promoting action also on epidermal keratinocytes.

[Test Example 6] Elastase Activity Inhibitory Action Test The ejaculation activity (sample 1) was tested for elastase activity inhibitory action as follows.

  A 0.2 mol / L Tris-HCl (pH 8.0) buffer solution in which a test sample (sample 1, sample concentration is shown in Table 6 below) was dissolved, or 0.2 mol / L Tris-HCl (No sample added) ( (pH 8.0) 50 μL of buffer solution and 50 μL of elastase solution (manufactured by Sigma) were mixed in a 96-well plate. Thereafter, a substrate solution (SIGMA, N-succinyl-Ala-Ala-Ala-p-nitroanilide, dissolved in 0.2 mol / L Tris-HCl (pH 8.0) buffer to a concentration of 0.4514 mg / mL) ) 100 μL was added and allowed to react at 25 ° C. for 15 minutes. After completion of the reaction, absorbance at a wavelength of 415 nm was measured using a spectrophotometer (BIO-TEK INSTRUMENTS, INC). Similarly, a blank test with no enzyme added was corrected. From the obtained results, the elastase activity inhibition rate (%) was calculated by the following formula.

Elastase activity inhibition rate (%) = {1− (A−B) / (C−D)} × 100
In the formula, A represents “absorbance at a wavelength of 415 nm when a test sample is added / enzyme added”, B represents “absorbance at a wavelength of 415 nm when a test sample is added / no enzyme is added”, and C is “no sample added / enzyme” D represents the “absorbance at a wavelength of 415 nm when no sample is added and no enzyme is added”.
The results are shown in Table 6.

  As shown in Table 6, it was confirmed that the Hihatsu extract (sample 1) has an excellent elastase activity inhibitory action.

Test Example 7 Type IV Collagen Production Promoting Action Test The above type of chickpea extract (Sample 1) was tested for type IV collagen production promoting action as follows.

Human normal skin fibroblasts (NB1RGB) were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS to a cell density of 1.6 × 10 ⁵ cells / mL, and then seeded at 100 μL per well in a 96-well plate and cultured overnight.

  After completion of the culture, the medium was removed, and 0.25% FBS-containing Dulbecco MEM medium to which a test sample (sample 1, sample concentration see Table 7 below) was added or 0.25% FBS-containing Dulbecco MEM medium to which no sample was added 150 μL was added to each well and cultured for 3 days. After completion of the culture, the amount of type IV collagen in the medium of each well was measured by ELISA using a monoclonal anti-type IV collagen antibody (mouse IgG, manufactured by Sigma). From the obtained results, the rate of production of type IV collagen (%) when the sample solution was added was calculated according to the following formula.

Type IV collagen production promotion rate (%) = A / B × 100
In the formula, A indicates “amount of type IV collagen when a test sample is added” and B indicates “amount of type IV collagen when no sample is added”.
The results are shown in Table 7.

  As shown in Table 7, it was confirmed that the Hihatsu extract (Sample 1) has an excellent type IV collagen production promoting action.

[Test Example 8] Epidermal hyaluronic acid production promoting action test The epidermis hyaluronic acid production promoting action was tested as follows for the above-mentioned Hihatsu extract (sample 1).

Human normal newborn skin epidermal keratinocytes (NHEK) were cultured using a growth medium for long-term culture of human normal epidermal keratinocytes (EpiLife-KG2), and then cells were collected by trypsin treatment. The collected cells were diluted with Epilife-KG2 medium to a cell density of 1 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured for 24 hours.

  After completion of the culture, 100 μL of Epilife-KG2 medium to which the test sample (sample 1, sample concentration is shown in Table 8 below) or the sample-free Epilife-KG2 medium was added was added to each well and cultured for 7 days. After the culture, the amount of hyaluronic acid in the medium of each well was measured by a sandwich method using hyaluronic acid binding protein (HABP). From the measurement results, the hyaluronic acid production promotion rate (%) was calculated by the following formula.

Hyaluronic acid production promotion rate (%) = A / B × 100
In the above formula, A represents “the amount of hyaluronic acid when the test sample is added”, and B represents “the amount of hyaluronic acid when no sample is added”.
The results are shown in Table 8.

  As shown in Table 8, it was confirmed that the Hihatsu extract (Sample 1) has an excellent hyaluronic acid production promoting action. Moreover, it was confirmed that the hyaluronic acid production promoting action of the hihatsu extract depends on the concentration of the extract.

[Test Example 9] Profilaggrin / Filagulin Production Promoting Action Test The prophylagrin / filaggulin production promoting action of the above-mentioned Hihatsu extract (Sample 1) was tested as follows.

Normal human neonatal epidermal keratinocytes (NHEK) were cultured at 37 ° C., 5% CO ₂ -95% air using a growth medium (EpiLife-KG2) for long-term culture of human normal epidermal keratinocytes in a 75 cm ² flask. The cells were collected by a conventional method. The obtained cells are diluted in Epilife-KG2 medium so as to have a cell density of 1.5 × 10 ⁵ cells / mL, then seeded at 2 mL each on a 6-well collagen-coated plate, and incubated at 37 ° C., 5% CO ₂ -95. The cells were cultured for 3 days under the condition of% air. After culturing, the medium was removed, and the Epilife-KG2 medium to which the test sample dissolved in 0.25% DMSO (sample 1, sample concentration as shown in Table 9 below) was added or the sample-free Epilife-KG2 medium was added to each well. 2 mL was added to each, and cultured for 5 days under the conditions of 37 ° C., 5% CO ₂ -95% air. After completion of the culture, total protein was prepared by a conventional method.

  The sample adjusted to 10 μg / row was developed by SDS-PAGE and transferred to a PVDF membrane. After blocking with PBS buffer containing 5% skim milk, anti-human filaggrin monoclonal antibody (Santa Cruz Biotechnology), biotin-labeled anti-mouse IgG (Amersham Biosciences, Whole Ab) and streptavidin-peroxidase complex (CALBIOCHEM) Made of 0.1% Tween 20 and 0.3% skimmed milk in PBS buffer, diluted 1000-fold, and sequentially reacted, and profilagline and filaggrin by luminescence of ECL Western blotting detection reagents (Amersham Biosciences). Was detected using an image photographing device (Bio-Rad Laboratories, ChemiDoc XRS Plus). The detected band was quantitatively measured with Image Lab Software version 2.0 (manufactured by Bio-Rad Laboratories).

  As a result, the profilaggrin production promoting effect of the sample was evaluated by using the value obtained by adding the net intensity (band intensity) of profilagrin and filaggrin in 10 μg of the protein prepared from the cells cultured with and without the sample added. The profilaggrin production promotion rate (%) was calculated based on the following formula.

Profilaggrin / filaggrin production promotion rate (%) = A / B × 100
In the formula, A represents “Net intensity at the time of sample addition (total value of profilagrin and filaggrin)”, and B represents “Net intensity at the time of no sample addition (control)”.
The results are shown in Table 9.

  As shown in Table 9, it was confirmed that the Hihatsu extract (Sample 1) has an excellent profilaggrin / filaggrin production promoting action. In addition, since filaggrin is produced by hydrolysis of profilagrin in the living body, the extract of baboon that has profilagrin production promoting action promotes the production of profilagrin and reduces the amount of profilagrin in the cell. By increasing it, the amount of filaggrin can also be increased, and as a result, it is considered to have an action of promoting filaggrin production.

[Test Example 10] Involucrin production promoting action test The incubulin production promoting action was tested as follows for the above-mentioned Hihatsu extract (Sample 1).

Human normal neonatal epidermal keratinocytes (NHEK) were used in a culture medium for long-term culture of human normal epidermal keratinocytes (EpiLife-KG2) in an 80 cm ² flask under conditions of 37 ° C. and 5% CO ₂ -95% air. And the cells were recovered by trypsin treatment. The collected cells are diluted with Epilife-KG2 medium to a cell density of 1.0 × 10 ⁵ cells / mL, and then seeded at 200 μL per well in a 48-well plate at 37 ° C., 5% CO ₂ -95. The cells were cultured overnight under the condition of% air.

After completion of the culture, the medium was removed, and 200 μL of Epilife-KG2 medium to which the test sample (sample 1, sample concentration is shown in Table 10 below) or the sample-free Epilife-KG2 medium was added was added to each well. ° _C., and cultured for 48 hours under the conditions of 5% CO 2 -95% air. After completion of the culture, the medium was removed, and the amount of involucrin expressed on the cell surface of the cells fixed on the plate was measured by ELISA using a monoclonal anti-human involucrin antibody (mouse IgG, manufactured by Sigma). From the obtained measurement results, the involucrin production promotion rate (%) was calculated by the following formula.

Involucrin production promotion rate (%) = A / B × 100
In the formula, A represents “absorbance at a wavelength of 405 nm when a test sample is added”, and B represents “absorbance at a wavelength of 405 nm when no sample is added”.
The results of the above test are shown in Table 10.

  As shown in Table 10, it was confirmed that the Hihatsu extract (Sample 1) has an excellent involucrin production promoting action.

  Hexosaminidase release inhibitor of the present invention, stem cell growth factor mRNA expression increase inhibitor, hydrogen peroxide cell disorder preventive / ameliorating agent, glutathione production promoter, elastase activity inhibitor, type IV collagen production promoter, hyaluronic acid Production promoters, profilagrin production promoters, filaggrin production promoters and involucrin production promoters are inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, various skin diseases associated with rough skin; Buckwheat, skin pigmentation; lung disease caused by smoking, cataract, various arteriosclerosis (ischemic heart disease, myocardial infarction, cerebral ischemia, cerebral infarction, etc.), aging with age, skin spots, etc. Pigmentation, neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, Huntington's chorea, etc.), diseases caused by oxidative stress such as canceration; Dull, texture loss, prevention of aging skin conditions such as reduced elasticity of the reduction and moisturizing function, can greatly contribute to the treatment or amelioration.

Claims (10)

  An inhibitor of hexosaminidase release, comprising an extract from Hihatsu as an active ingredient.   An inhibitor of stem cell growth factor mRNA expression increase, comprising an extract from baboon as an active ingredient.   A prophylactic / ameliorating agent for hydrogen peroxide cell damage, characterized by containing an extract from baboon as an active ingredient.   Glutathione production promoter characterized by containing an extract from Hihatsu as an active ingredient.   An elastase activity inhibitor characterized by containing an extract from baboon as an active ingredient.   A type IV collagen production promoter characterized by containing an extract from Hihatsu as an active ingredient.   A hyaluronic acid production promoter characterized by containing an extract from hihitsu as an active ingredient.   A profilaggrin production promoter characterized by containing an extract from Hihatsu as an active ingredient.   A filaggrin production promoter characterized by containing an extract from Hihatsu as an active ingredient.   An involucrin production promoter characterized by containing an extract from baboon as an active ingredient.