JP5612267B2 - Skin cosmetics - Google Patents

Description

  The present invention relates to an antioxidant, an anti-inflammatory agent, a whitening agent, an anti-aging agent, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a skin cosmetic, and a food and drink.

In recent years, active oxygen has attracted attention as a factor that particularly oxidizes biological components, and its adverse effect on living organisms has become a problem. Active oxygen is generated in the process of energy metabolism in living cells. Examples of active oxygen include superoxide (that is, superoxide anion generated by one-electron reduction of oxygen molecules: .O ₂ ⁻ ), hydrogen peroxide (H ₂ O ₂ ), hydroxy radical (.OH), singlet oxygen ( ¹ O ₂ ) and the like. These active oxygens are essential for the phagocytic sterilization mechanism and play an important role in removing viruses and cancer cells.

  However, excessive production of active oxygen attacks in vivo molecules constituting membranes and tissues in the living body and induces various diseases. Normally, superoxide produced in vivo and used as a starting material for other active oxygen is sequentially eliminated by the catalytic action of superoxide dismutase (SOD) contained in the cells. Is excessive or when the SOD action is reduced, superoxide elimination is insufficient, and the superoxide concentration increases, which may cause tissue damage such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke Cause cataracts, wrinkles, diabetes, arteriosclerosis, stiff shoulders, coldness.

  In particular, since the skin is directly affected by environmental factors such as ultraviolet rays, it is an organ that easily generates superoxide, and therefore, by increasing the concentration of superoxide, for example, the biological tissue such as collagen is decomposed and denatured or It is thought to crosslink and oxidize fats and oils to produce lipid peroxides that damage cells, and the damage caused by active oxygen is the cause of aging such as skin wrinkle formation and skin elasticity reduction (See Non-Patent Document 1). Therefore, by inhibiting and suppressing the generation of active oxygen and in vivo radicals, skin aging such as wrinkle formation and reduced elasticity, tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataracts, diabetes, arteries It is considered that various disorders involving active oxygen such as hardening, stiff shoulders, and coldness can be prevented, treated or improved.

  Therefore, attempts have been made to obtain active oxygen scavenging substances, radical scavenging substances, etc. from natural products advantageous in terms of safety. Extracts from Brassica family Brassica (Patent Documents) 1) and the like are known.

  There are various causes and onset mechanisms of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and various other skin diseases with rough skin. Known causes include tumor necrosis factor (hereinafter also referred to as “TNF-α”) produced mainly by macrophages, hyaluronidase activity enhancement, and histamine release. Yes.

  TNF-α was found as a factor that necrotizes tumors, but is recently considered to be a cytokine that plays a mediator role not only for tumors but also for regulating the function of normal cells. TNF-α plays an important role in the process from the onset to the end of inflammation, but its persistent and excessive production causes damage to tissues including the skin, and causes systemic fever and cachycecia. And cause worsening of inflammation. As such inflammation, for example, chronic inflammatory diseases such as rheumatoid arthritis and osteoarthritis are representative. Therefore, in pathological inflammation, it is important to suppress excessive production of TNF-α. Known examples of such a TNF-α production inhibitory effect include perilla extract (see Non-Patent Document 2), Amaryllidaceae alkaloids ricolin and licocidinol (see Non-Patent Document 3).

  Hyaluronidase is a hydrolase of hyaluronic acid. Hyaluronate that maintains affinity for body tissues is decomposed by ultraviolet rays, oxygen, and the like in a water-containing system, and the water retention effect decreases with a decrease in molecular weight. In addition, hyaluronic acid exists as an intercellular tissue in vivo and is also involved in blood vessel permeability. Furthermore, although hyaluronidase exists in mast cells, it is released by degranulation caused by its activation and acts as an inflammatory chemical mediator. Therefore, by inhibiting the activity of hyaluronidase, enhancement of moisture retention and prevention / reduction of inflammation are expected. As what has such a hyaluronidase activity inhibitory effect, the extract (refer patent document 2), the Fuji tea extract (refer patent document 3), etc. of the genus Osbeckia are known, for example.

  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, it is considered that by suppressing the release of hexosaminidase, the release of histamine can also be suppressed at the same time, which is effective for preventing, treating or improving inflammatory diseases. As a plant extract having such a hexosaminidase release inhibitory effect, an extract from the above-mentioned Fuji tea (see Patent Document 3) and the like are known.

  Melanin also plays a role in protecting the body from ultraviolet rays in the skin, but overproduction and uneven accumulation cause skin darkening and spots. In general, melanin is converted from tyrosine to dopa and from dopa to dopaquinone by the action of the enzyme tyrosinase biosynthesized in pigment cells, and then formed through intermediates such as 5,6-dihydroxyindophenol. ing. Therefore, to prevent, treat, or improve skin darkness (skin pigmentation), spots, buckwheat, etc., inhibit the activity of tyrosinase involved in the production of melanin, or suppress the production of melanin. Can be considered.

  Conventionally, for the prevention, treatment or improvement of skin pigmentation, stains, buckwheat, etc., a treatment using a whitening agent containing a chemically synthesized product such as hydroquinone as an active ingredient has been performed. However, chemically synthesized products such as hydroquinone may cause side effects such as skin irritation and allergies. Therefore, development of a whitening agent comprising a highly safe natural raw material as an active ingredient is desired. Examples of those having a tyrosinase activity inhibitory action include wisteria tea extract (see Patent Document 4), Yanagitade extract ( Patent Document 5) is known. Moreover, as what has a melanin production inhibitory effect, the extract (refer patent document 6) from the root part of castor bean (refer patent document 6), the extract (refer patent document 7) etc. which belong to the south urea (Saussurea) genus are known, for example. Yes.

  Conventional whitening agent development has been focused on tyrosinase, which is the rate-limiting enzyme for melanin production. Recently, as a cytokine that is produced from epidermal keratinocytes after ultraviolet (UV-B) irradiation and activates melanocytes, -Melanocyte-stimulating hormone (α-MSH), endothelin-1 (ET-1), nitric oxide (NO), etc. are known, and the production of melanin is suppressed by blocking the signal transduction system involved. Various agents that lead to whitening effects have been actively developed.

  In patients with cardiovascular diseases (for example, myocardial infarction, hypertension, ischemic heart disease, etc.) or renal diseases (for example, acute renal failure, etc.), blood endothelin-1 (ET-1) concentration increases. Since it is known that, by inhibiting excessive secretion of endothelin-1 (ET-1), that is, by suppressing the increase in the expression of endothelin-1 mRNA, the prevention / treatment effect of these diseases can be expected. Conceivable.

  Based on this idea, chamomile extract, artea extract (see Non-Patent Document 4) and the like are known as herbal medicines that inhibit the action of endothelin-1 (ET-1) on melanocytes, and epidermal keratinocytes are known. As herbal medicines that suppress endothelin-1 (ET-1) production from cucumber, diu extract (see non-patent document 4), β-glycyrrhetinic acid stearyl (see patent document 8) and the like are known.

  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 precursor cells, granulocyte / macrophage precursor cells, pigment cells and the like. In addition, it is known that expression of SCF is enhanced by a spot site, ultraviolet irradiation, or the like (see Non-Patent Document 5).

  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 the binding site, released from the cell membrane, and bound to the SCF receptor of the pigment cell, thereby promoting proliferation of the pigment cell. Furthermore, SCF is a bone marrow 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 blasts (see Non-Patent Document 6).

  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 the expression of SCF mRNA 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 mRNA 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 such an idea, for example, an extract from Arnica is known as one having an action of suppressing the production / release of SCF (see Patent Document 9).

  It is known that α-MSH as a cytokine that activates ACTH and melanocytes, which are secreted from skin epidermis cells and are involved in the production of melanin, is produced using proopiomelanocortin (POMC) as a precursor. . Therefore, by suppressing the production of POMC, that is, by suppressing the increase in the expression of POMC mRNA, it is possible to suppress the production of melanin as a result, and to prevent or improve pigmentation, blemishes, freckles, etc. after sunburn It is thought that you can.

  Conventionally, what has the effect | action which suppresses the expression of POMC is known, for example, panthenol, pyridoxine hydrochloride, nicotinamide, and the like (see Patent Document 10).

  Various factors are involved in complex skin aging. For the purpose of preventing or delaying skin aging, for example, active research has been conducted on the inhibition of the activity of matrix proteases present in the skin and involved in the degradation of the skin.

  The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as elastin and collagen that are outside these cells and support the skin structure. In young skin, moisture retention, flexibility, elasticity and the like are ensured by maintaining the constancy of the interaction between these skin tissues, and the skin is maintained in a fresh and fresh state.

  However, elastin, which is a major component of the extracellular matrix, degrades and deteriorates when affected by certain external factors such as UV irradiation, drastic air drying, excessive skin washing, and so on. In addition, the production amount of collagen is reduced, and the elasticity is lowered due to crosslinking. As a result, the moisturizing function and elasticity of the skin are deteriorated, 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 associated with skin aging, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like, are associated with a decrease and degeneration of dermal matrix components such as collagen and elastin.

  In recent years, the involvement of matrix metalloproteinases (MMPs) has been pointed out as factors that induce changes associated with skin aging. Among these MMPs, matrix metalloproteinase-1 (MMP-1) is known as an enzyme that degrades collagen, which is a major component of the dermal extracellular matrix of skin, but its expression is greatly increased by irradiation with ultraviolet rays. However, it is thought to contribute to the decrease / denaturation of collagen, and to be a major factor such as the formation of wrinkles on the skin and the decrease in elasticity. Therefore, inhibiting the activity of MMP-1 is important in preventing and improving skin aging symptoms. As what has such an MMP-1 inhibitory action, for example, an extract from Himalayan cherry (see Patent Document 11), an extract from Ginger family Genghis Kaba Sumner or Mulberry family Ficus nerifolia (see Patent Document 12) Etc. are known.

  Skin structure is roughly divided into epidermis, basement membrane, dermis, and subcutaneous tissue, and basement membrane plays an extremely important role in maintaining skin structure and homeostasis. Therefore, when the structure of the basement membrane changes, skin aging symptoms such as wrinkles and sagging appear. In particular, when the production amount of type IV collagen, which is a main component of the basement membrane, decreases, the structure of the basement membrane changes, resulting in skin aging symptoms such as wrinkles and sagging.

  The dermis of the skin is composed of fibroblasts and extracellular matrices such as elastin, hyaluronic acid, and type I collagen that are outside these cells and support the skin structure. For this reason, the dermis plays an important role in ensuring moisture retention, flexibility, and elasticity necessary for skin tension and gloss. Therefore, when the production of type I collagen decreases due to the influence of certain external factors such as ultraviolet irradiation and drying, aging, etc., the structure of the dermis changes, the moisturizing function decreases, and the elasticity of the skin It begins to show skin aging symptoms such as reduction, reduction in tension and gloss.

  Conventionally, what has a collagen production promoting action is a plant selected from the group consisting of a quince extract (see Patent Document 4), hydrolyzed casein, beech buds, erythrina, soluble eggshell membranes, cuckoo, western ivy and Animal-derived extracts and the like are known (see Patent Document 13).

  When the skin is irradiated with ultraviolet rays, the cells of the skin are damaged or cell death is caused, the skin loses its elasticity and elasticity, and exhibits aging symptoms such as rough skin and wrinkles. Therefore, prevention / improvement of skin aging can be expected by suppressing / recovering damage (for example, cell damage, cell death, etc.) caused by ultraviolet irradiation. For example, an oil-soluble licorice extract (see Patent Document 14) is known as one having a damage recovery effect by ultraviolet irradiation.

  In recent years, body fat has increased due to lifestyle habits such as satiety and lack of exercise, and obesity has increased. Such an increase in obesity is seen not only in humans but also in pets and livestock. Obesity is a cause of adult diseases such as hyperlipidemia and arteriosclerosis, which is not only a cosmetic problem but also a major problem in health.

  In order to break down fat in the living body, the role of cyclic AMP is important. Cyclic AMP activates lipase present in the living body, and fat is decomposed into fatty acid and glycerol by the activated lipase. However, when cyclic AMP phosphodiesterase is activated, degradation of cyclic AMP is induced and lipase activation is inhibited. Therefore, it is considered that by inhibiting the activity of cyclic AMP phosphodiesterase, the amount of cyclic AMP in the cell can be increased and the decomposition of fat can be promoted. A peanut astringent skin extract (see Patent Document 15) or the like is known as having such a cyclic AMP phosphodiesterase activity inhibitory action.

JP 2003-81848 A Japanese Patent Laid-Open No. 2003-55242 JP 2003-12532 A JP 2002-370962 A JP 2004-83488 A JP 2001-213757 A JP 2002-201222 A JP 2004-300048 A JP 2008-169118 A JP 2007-176810 A JP 2003-176232 A JP 2003-176230 A JP 2004-18471 A JP 2004-250368 A JP 2004-26719 A

"Fragrance Journal Extra Number", 1995, No.14, p.156 “Inflammation”, 1993, Vol. 13, No. 4, p.337-340 "Pharmaceutical Journal", 2001, Vol.121, No.2, p.167-171 “Fragrance Journal”, 2000, Vol. 28, No. 9, p. 65-71 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

  The present invention relates to an antioxidant, an anti-inflammatory agent, a whitening agent, an anti-aging agent, an anti-obesity agent, and a cyclic AMP phosphodiesterase activity inhibitor comprising a highly safe natural extract and / or a component derived from a natural extract as an active ingredient. It is another object of the present invention to provide a skin cosmetic and a food or drink containing the natural extract and / or the natural extract-derived component.

  In order to achieve the above object, an antioxidant, an anti-inflammatory agent, a whitening agent, an anti-aging agent, an anti-obesity agent or a cyclic AMP phosphodiesterase activity inhibitor contains an extract from loquat flower as an active ingredient It is characterized by that. Moreover, the skin cosmetics or food / beverage products of this invention mix | blended the extract from the flower part of a loquat.

  In the antioxidant of the present invention, the extract from the loquat flower part preferably has an active oxygen scavenging action and / or a radical scavenging action. In the anti-inflammatory agent of the present invention, the loquat flower part It is preferable that the extract from Tumor Necrosis Factor (TNF-α) production suppressive action, hyaluronidase activity inhibitory action and hexosaminidase release inhibitory action have one or more actions selected from the group consisting of In the whitening agent of the present invention, the extract from the flower part of loquat has a tyrosinase activity inhibitory action, a melanin production inhibitory action, an endothelin-1 mRNA expression rise inhibitory action, a stem cell growth factor (SCF) mRNA expression rise inhibitory action and a pro It preferably has one or more actions selected from the group consisting of opiomelanocortin (POMC) mRNA expression increase inhibitory action. In addition, in the anti-aging agent of the present invention, the extract from the flower part of loquat has a matrix metalloproteinase-1 (MMP-1) activity inhibitory action, a type I collagen production promoting action, a type IV collagen production promoting action, and It preferably has one or more actions selected from the group consisting of damage recovery action by ultraviolet irradiation.

  According to the present invention, an extract from loquat that is a natural product is contained as an active ingredient, and a highly safe antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitors, skin cosmetics and foods and drinks can be provided.

Hereinafter, an embodiment of the present invention will be described.
[Antioxidants, anti-inflammatory agents, whitening agents, anti-aging agents, anti-obesity agents, cyclic AMP phosphodiesterase activity inhibitors]
The antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention contains an extract from loquat flower as an active ingredient.

  Here, in the present invention, the “extract” is obtained by drying an extract obtained from a flower part of loquat (scientific name: Eriobotrya japonica), a diluted or concentrated solution of the extract, and the extract. Or any of these crude or purified products.

  Biwa (Eriobotrya japonica) is an evergreen tree belonging to the genus Biwa, and is native to the southwestern part of China. It is cultivated in Nagasaki, Chiba, Kagoshima, etc., and can be easily obtained from these areas. The constituent part of loquat used as an extraction raw material is a flower part. Here, the "flower" generally refers to the whole organ involved in sexual reproduction of a seed plant, and is composed of a flower leaf that is a leaf deformation and a flower axis that is a stem deformation. This includes organs such as pupae, petals, stamens, and heart skin. The “floral part” used as an extraction raw material in the present invention includes all of the organs involved in sexual reproduction of seed plants, as well as parts thereof, such as flower leaves, flower coats (buds and corolla), corolla, petals, etc. Is also included.

  The extract from the flower part of loquat 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. At this time, the extraction raw material may be dried in the sun or by 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, the extraction raw material can be efficiently extracted with a polar solvent.

  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 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 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. 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 an active ingredient of an antioxidant, an anti-inflammatory agent, a whitening agent, an anti-aging agent, an anti-obesity agent or a cyclic AMP phosphodiesterase activity inhibitor as it is, but a concentrated solution or a dried product. It is easier to use.

  The extract from the loquat flower part has a unique odor and can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. Since it is not used in a large amount when blended into foods or foods, there is no practical problem even if it is not purified. In addition, since the extract from the loquat flower part has a light brown color, and its color tone is thin, even if the amount of the extract from the loquat flower part is increased in the skin cosmetics or food and drink, the skin There is no risk of affecting the color of cosmetics or food and drink.

  Since the extract from the flower part of loquat obtained as described above has an antioxidant action, anti-inflammatory action, whitening action, anti-aging action, anti-obesity action or cyclic AMP phosphodiesterase activity inhibitory action, Each action can be used as an active ingredient of an antioxidant, an anti-inflammatory agent, a whitening agent, an anti-aging agent, an anti-obesity agent or a cyclic AMP phosphodiesterase inhibitor.

  The antioxidant effect of the extract from the flower part of loquat is exhibited based on, for example, an active oxygen scavenging action and / or a radical scavenging action. However, the antioxidant action which the extract from the flower part of loquat has is not limited to the antioxidant action exhibited based on these actions.

  The anti-inflammatory action of the extract from the flower part of loquat is, for example, one kind selected from the group consisting of tumor necrosis factor (TNF-α) production inhibitory action, hyaluronidase activity inhibitory action, and hexosaminidase release inhibitory action, or It is demonstrated based on two or more actions. However, the anti-inflammatory action which the extract from the flower part of loquat has is not limited to the anti-inflammatory action exhibited based on these actions.

  The whitening action of the loquat extract has, for example, tyrosinase activity inhibitory action, melanin production inhibitory action, endothelin-1 mRNA expression rise inhibitory action, stem cell growth factor (SCF) mRNA expression rise inhibitory action, and proopiomelanocortin ( POMC) It is exhibited based on one or more actions selected from the group consisting of the suppressive action on the increase in mRNA expression. However, the whitening action of the extract from the loquat flower part is not limited to the whitening action exhibited based on these actions.

  Anti-aging effects of extracts from loquat flowers include, for example, matrix metalloproteinase-1 (MMP-1) activity inhibition action, type I collagen production promotion action, type IV collagen production promotion action, and damage recovery by ultraviolet irradiation. It is exhibited based on one or more actions selected from the group consisting of actions. However, the anti-aging action which the extract from a loquat flower part has is not limited to the anti-aging action exhibited based on these actions.

  The anti-obesity effect of the extract from the flower part of loquat is exhibited based on, for example, a cyclic AMP phosphodiesterase activity inhibitory action. However, the anti-obesity action that the extract from the flower part of loquat has is not limited to the anti-obesity action exhibited based on this action.

  In addition, the extract from the flower part of loquat is an active oxygen scavenging action, radical scavenging action, tumor necrosis factor (TNF-α) production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, tyrosinase activity inhibitory action. , Melanin production inhibitory action, endothelin-1 mRNA expression increase inhibitory action, stem cell growth factor (SCF) mRNA expression increase inhibitory action, proopiomelanocortin (POMC) mRNA expression increase inhibitory action, matrix metalloproteinase-1 (MMP-1) activity inhibitory action Since it has an action, a type I collagen production promotion action, a type IV collagen production promotion action or a damage recovery action by ultraviolet irradiation, the active oxygen scavenger, radical scavenger, tumor necrosis factor (TNF-α) ) Production inhibitor, Hyaluronidase activity inhibitor, Hexo Saminidase release inhibitor, tyrosinase activity inhibitor, melanin production inhibitor, endothelin-1 mRNA expression increase inhibitor, stem cell growth factor (SCF) mRNA expression increase inhibitor, proopiomelanocortin (POMC) mRNA expression increase inhibitor, matrix metalloproteinase -1 (MMP-1) activity inhibitor, type I collagen production promoter, type IV collagen production promoter or an active ingredient of a damage recovery agent by ultraviolet irradiation.

  In addition, the extract from the flower part of loquat is used to prevent or treat diseases caused by increased expression of endothelin-1 mRNA (for example, circulation of myocardial infarction, hypertension, etc.) using its endothelin-1 mRNA expression increase inhibitory action. It can also be used as an active ingredient of a prophylactic / therapeutic agent for systemic diseases.

  Furthermore, the extract from the flower part of loquat is used for the preventive / therapeutic agent for diseases caused by the increased expression of SCF mRNA (for example, the preventive / therapeutic agent for myelodysplastic syndrome, It can also be used as an active ingredient of prophylactic leukemia preventive / therapeutic agents, antitumor agents and the like.

  Furthermore, the extract from the flower part of loquat is effective as a prophylactic / therapeutic agent for diseases caused by increased expression of POMC mRNA (for example, stress pruritus etc.) using its inhibitory effect on the increase in POMC mRNA expression. It can also be used as a component.

  The antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention may consist only of an extract from the flower part of loquat, It may be a formulation of an extract from the flower part.

  Extracts from loquat flowers can be used in any form of powder, granules, tablets, liquids, etc. according to conventional methods using pharmaceutically acceptable carriers such as dextrin and cyclodextrin. It can be formulated into a dosage form. 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. Moreover, the extract from a flower part of a loquat can be mix | blended and used for other compositions (for example, skin cosmetics, food-drinks, etc.).

  The antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention may be antioxidant, anti-inflammatory, whitening, Other natural extracts having an aging action, an anti-obesity action or a cyclic AMP phosphodiesterase activity inhibiting action can be blended and used as an active ingredient.

  The administration method of the antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention generally includes transdermal administration, oral administration, etc. A suitable method for the prevention / treatment or the like may be appropriately selected depending on the type.

  In addition, the dosage of the antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention is also the type of disease, severity, individual differences among patients, administration method The dose may be appropriately increased or decreased depending on the administration period.

  Antioxidant of the present invention, through the active oxygen scavenging action and / or radical scavenging action of extracts from loquat flowers, skin aging such as wrinkle formation and reduced elasticity, tissues such as rheumatoid arthritis and Behcet's disease Various disorders involving active oxygen such as disorder, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, and coldness can be prevented and improved. However, the antioxidant of the present invention can be used for all purposes that are meaningful for exhibiting an active oxygen scavenging action and / or a radical scavenging action in addition to these uses.

  The anti-inflammatory agent of the present invention is one selected from the group consisting of a tumor necrosis factor (TNF-α) production inhibitory action, a hyaluronidase activity inhibitory action, and a hexosaminidase release inhibitory action possessed by an extract from the flower part of loquat Alternatively, through two or more kinds of actions, contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other various inflammatory skin diseases associated with rough skin can be prevented and improved. However, the anti-inflammatory agent of the present invention is one or two selected from the group consisting of tumor necrosis factor (TNF-α) production inhibitory action, hyaluronidase activity inhibitory action, and hexosaminidase release inhibitory action in addition to these uses. It can be used for all purposes that are meaningful for exerting more than a kind of action.

  The whitening agent of the present invention comprises tyrosinase activity inhibitory action, melanin production inhibitory action, endothelin-1 mRNA expression rise inhibitory action, stem cell growth factor (SCF) mRNA expression rise inhibitory action and proopiomelanocortin possessed by extracts from loquat flowers. (POMC) It is possible to prevent or improve pigmentation such as darkening of the skin, spots and freckles through one or more actions selected from the group consisting of the action of suppressing the increase in mRNA expression. However, the whitening agent of the present invention has a tyrosinase activity inhibitory action, a melanin production inhibitory action, an endothelin-1 mRNA expression rise inhibitory action, a stem cell growth factor (SCF) mRNA expression rise inhibitory action, and proopiomelanocortin (POMC) in addition to these uses. ) It can be used for all purposes that are meaningful in exerting one or more kinds of actions selected from the group consisting of the action of suppressing the increase in mRNA expression.

  The anti-aging agent of the present invention is a matrix metalloproteinase-1 (MMP-1) activity inhibitory action, an action to promote type I collagen production, an action to promote type IV collagen production, and damage caused by ultraviolet irradiation. Skin aging symptoms and the like can be prevented and ameliorated through one or more actions selected from the group consisting of recovery actions. However, the anti-aging agent of the present invention has a matrix metalloproteinase-1 (MMP-1) activity inhibitory action, a type I collagen production promoting action, a type IV collagen production promoting action and a damage recovery action by ultraviolet irradiation in addition to these uses. It can be used for all applications that are meaningful for exhibiting one or more actions selected from the group consisting of:

  The anti-obesity agent of the present invention can promote the production of cyclic AMP through the cyclic AMP phosphodiesterase inhibitory action possessed by the extract from the flower of loquat and can decompose adipocytes. As a result, obesity In addition, various diseases such as arteriosclerosis, diabetes, and metabolic syndrome can be prevented and improved. However, the anti-obesity agent of the present invention can be used for all purposes other than these uses that are meaningful for exerting a cyclic AMP phosphodiesterase inhibitory action.

  The cyclic AMP phosphodiesterase inhibitor of the present invention can promote the production of cyclic AMP and promote the degradation of adipocytes through the cyclic AMP phosphodiesterase inhibitory action of the extract from the flower part of loquat. As a result, various diseases such as obesity, accompanying arteriosclerosis, diabetes, and metabolic syndrome can be prevented and improved. However, the cyclic AMP phosphodiesterase inhibitor of the present invention can be used for all purposes other than these uses that are meaningful for exerting a cyclic AMP phosphodiesterase inhibitory action.

[Skin cosmetic]
Extracts from loquat flowers have anti-oxidant, anti-inflammatory, whitening, anti-aging, anti-obesity, or cyclic AMP phosphodiesterase activity inhibitory effects. Or since it is excellent in safety | security, it is suitable for mix | blending with skin cosmetics.

  In this case, the skin cosmetic may be blended with the extract from the loquat flower part as it is, or an antioxidant, an anti-inflammatory agent, a whitening agent formulated from the extract from the loquat flower part, You may mix | blend an anti-aging agent, an anti-obesity agent, or a cyclic AMP phosphodiesterase activity inhibitor.

  Extract from loquat flower part, or antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor formulated from the extract from loquat flower part By blending, it is possible to impart an antioxidant action, an anti-inflammatory action, a whitening action, an anti-aging action, an anti-obesity action or a cyclic AMP phosphodiesterase activity inhibiting action to the skin cosmetic.

  There are no particular limitations on the type of skin cosmetic that can be blended with the extract from the flower part of loquat, and examples thereof include ointments, creams, emulsions, lotions, packs, foundations and the like.

  When blending extracts from loquat flowers into skin cosmetics, the blending amount can be adjusted as appropriate according to the type of skin cosmetics. It is about 0.0001 to 10% by mass in terms of conversion, and a particularly suitable blending ratio is about 0.001 to 1% by mass in terms of a standard extract.

  The skin cosmetic of the present invention is usually used as long as it does not interfere with the antioxidant, anti-inflammatory, whitening, anti-aging, anti-obesity or cyclic AMP phosphodiesterase activity inhibiting action of the extract from the loquat flower. Main ingredients, auxiliaries or other components used in the manufacture of skin cosmetics such as astringents, bactericides / antibacterial agents, UV absorbers, moisturizers, cell activators, anti-inflammatory / anti-allergic agents, antioxidant / active oxygen A remover, fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances and the like can be used in combination. By using together, it becomes a more general product, and the synergistic action between the above-mentioned components used in combination may bring about a superior use effect than would normally be expected.

  The skin cosmetic of the present invention has high safety and is selected from the group consisting of an antioxidant action, an anti-inflammatory action, a whitening action, an anti-aging action, an anti-obesity action and a cyclic AMP phosphodiesterase activity inhibiting action. Through one or more actions, skin aging such as wrinkle formation and reduced elasticity, tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, coldness, etc. Various disorders involving active oxygen; contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other inflammatory skin diseases with rough skin; pigmentation of skin darkening, spots, freckles, etc .; skin aging symptoms Obesity, arteriosclerosis associated with obesity, diabetes, metabolic syndrome and the like can be prevented, treated or improved.

[Food and Drink]
The extract from loquat has antioxidant, anti-inflammatory, whitening, anti-aging, anti-obesity, or cyclic AMP phosphodiesterase activity inhibitory effects, such as being digested in the digestive tract It is confirmed that it is not a thing, and since it is excellent also in safety, it is suitable for blending in food and drink.

  In this case, the extract from loquat flower part may be blended as it is, or the antioxidant, anti-inflammatory agent, whitening agent, anti-aging agent, anti-obesity formulated from the extract from loquat flower part An agent or a cyclic AMP phosphodiesterase activity inhibitor may be added.

  Eating or drinking antioxidants, anti-inflammatory agents, whitening agents, anti-aging agents, anti-obesity agents or cyclic AMP phosphodiesterase activity inhibitors formulated from loquat flower parts or loquat flower parts extracts When blended into products, the amount of active ingredients in them can be changed as appropriate in consideration of the purpose of use, symptoms, gender, etc. It is preferable that the daily intake of the extract is about 1 to 1000 mg.

  The food / beverage product of the present invention may be a blend of an extract from the loquat flower part in any food / beverage product that does not interfere with its activity, and the extract from the loquat flower part as a main component. It may be a dietary supplement.

  In producing the food and drink of the present invention, for example, sugars such as dextrin and starch; proteins such as gelatin, soy protein and corn protein; amino acids such as alanine, glutamine and isoleucine; polysaccharides such as cellulose and gum arabic Any food and drink of any shape can be obtained by adding any auxiliary agent such as oils and fats such as soybean oil and medium chain fatty acid triglycerides.

  The foods and drinks that can be blended with the extract from the flower portion of loquat are not particularly limited, but specific examples thereof include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (concentrated stock solutions of these drinks) And ice cream, ice sherbet, shaved ice, etc .; noodles such as buckwheat, udon, harusame, gyoza skin, cucumber skin, Chinese noodles, instant noodles; rice cake, chewing gum, candy, gum, Chocolate, tablet confectionery, snack confectionery, biscuits, jelly, jam, cream, baked confectionery and other confectionery; fish and livestock processed foods such as kamaboko, ham, sausage; dairy products such as processed milk and fermented milk; salad oil, tempura oil, Fats and oils processed foods such as margarine, mayonnaise, shortening, whipped cream, dressing; seasonings such as sauces and sauces; Various types of health / nutritional supplements; tablets, capsules, drinks, etc., and these foods and drinks are mixed with extracts from the flower parts of loquat Sometimes, auxiliary materials and additives that are usually used can be used in combination.

  The antioxidants, anti-inflammatory agents, whitening agents, anti-aging agents, anti-obesity agents, cyclic AMP phosphodiesterase activity inhibitors, skin cosmetics or foods and beverages of the present invention are suitably applied to humans. However, it can be applied to animals other than humans as long as the respective effects are 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] Production of loquat flower part water extract 300 ml of water was added to 30 g of dried loquat flower part, and the mixture was heated and extracted at 80 ° C for 2 hours with a reflux extractor and filtered while hot. The residue was further extracted in the same manner. The obtained extracts were combined, concentrated under reduced pressure, and further dried to obtain 6.2 g of loquat flower part water extract (Sample 1).

[Production Example 2] Production of 30% ethanol extract of loquat flower part In the same manner as in Production Example 1 except that 300 mL of 30% ethanol (volume ratio of water to ethanol = 7: 3) was used as the extraction solvent, 5.6 g of 30% ethanol extract of flower part was obtained (sample 2).

[Production Example 3] Production of 80% ethanol extract of loquat flower part In the same manner as in Production Example 1, except that 300 mL of 80% ethanol (volume ratio of water to ethanol = 1: 4) was used as the extraction solvent. The flower part 80% ethanol extract 4.4g was obtained (sample 3).

[Test Example 1] Superoxide scavenging action test (NBT method)
About the loquat flower part extract (samples 1-3) obtained by manufacture examples 1-3, the superoxide elimination action was tested as follows.

In a test tube, 2.4 mL of 0.05 M Na ₂ CO ₃ buffer (pH 10.2), and 3 mM xanthine, 3 mM EDTA, bovine serum albumin solution and 0.75 mM NBT (nitroblue tetrazolium) were added. 1 mL each was added, 0.1 mL of the sample solution (Samples 1 to 3) was added thereto, and the mixture was allowed to stand at 25 ° C. for 10 minutes. After standing, a xanthine oxidase solution as an enzyme solution was added, stirred rapidly, and allowed to react at 25 ° C. for 20 minutes. Thereafter, 0.1 mL of 6 mM copper chloride was added to stop the reaction, and the absorbance at a wavelength of 560 nm was measured.

  Even when the enzyme solution was not added, the same operation and absorbance were measured, and the same measurement was performed when distilled water was added without adding the sample solution. From the obtained results, the superoxide elimination rate (%) was calculated by the following formula.

Superoxide erasure rate (%) = {1- (AB) / (CD)} × 100
In the formula, A represents “absorbance when enzyme solution is added / sample solution added”, B represents “absorbance when enzyme solution is not added / sample solution is added”, and C is “when enzyme solution is added / sample solution is not added” "D" represents "absorbance when no enzyme solution is added / sample solution is not added".

The concentration of the sample solution was decreased stepwise to measure the superoxide elimination rate, and the sample concentration IC ₅₀ (μg / mL) at which the superoxide elimination rate was 50% was determined by interpolation.
The results are shown in Table 1.

  As shown in Table 1, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent superoxide scavenging action (reactive oxygen scavenging action). It was also confirmed that the degree of superoxide scavenging action can be adjusted by the concentration of loquat flower part extract.

[Test Example 2] Radical scavenging action test The loquat flower part extracts (Samples 1 to 3) obtained in Production Examples 1 to 3 were tested for radical scavenging action as follows.

After 3 mL of the sample solution (Samples 1 to 3) was added to 3 mL of 1.5 × 10 ⁻⁴ M DPPH (diphenyl-p-picrylhydrazyl) ethanol solution, the solution was tightly stoppered and shaken and allowed to stand for 30 minutes. After standing, the absorbance at a wavelength of 520 nm was measured. As a control, the same operation was performed using only the solvent in which the sample was dissolved instead of the sample solution, and the absorbance at a wavelength of 520 nm was measured. Further, as a blank, after adding 3 mL of the sample solution to ethanol, the absorbance at a wavelength of 520 nm was measured immediately. From the obtained results, the radical scavenging rate (%) was calculated by the following formula.

Radical scavenging rate (%) = {1- (BC) / A} × 100
In the formula, A represents “absorbance of control”, B represents “absorbance upon addition of sample solution”, and C represents “absorbance of blank”.

The radical scavenging rate was measured by gradually decreasing the concentration of the sample solution, and the sample concentration IC ₅₀ (μg / mL) at which the radical scavenging rate was 50% was determined by interpolation.
The results are shown in Table 2.

  As shown in Table 2, the loquat flower part extract (samples 1 to 3) was confirmed to have an excellent radical scavenging action. Moreover, it was confirmed that the degree of radical scavenging action can be adjusted by the concentration of loquat flower part extract.

[Test Example 3] TNF-α production inhibitory action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for TNF-α production inhibitory action as follows.

After culturing mouse macrophage cells (RAW264.7) using Dulbecco's MEM medium containing 10% FBS, the cells were collected with a cell scraper. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS to a cell density of 1.0 × 10 ⁶ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured for 4 hours.

  After completion of the culture, the medium was removed, and 100 μL of a sample solution dissolved in Dulbecco MEM containing 10% FBS containing a final concentration of 2% DMSO (Samples 1 to 3, see Table 3 below for the sample concentration) was added to each well. 100 μL of lipopolysaccharide (LPS, E. coli 0111; B4, manufactured by DIFCO) dissolved in Dulbecco MEM containing 10% FBS at a final concentration of 1 μg / mL was added and cultured for 24 hours. After completion of the culture, the amount of TNF-α in the culture supernatant of each well was measured using a sandwich ELISA method, and the TNF-α production inhibition rate (%) was calculated from the measurement result according to the following formula.

TNF-α production inhibition rate (%) = {(B−A) / B} × 100
In the formula, A represents “a TNF-α amount when a sample solution is added”, and B represents a “a TNF-α amount when a sample solution is not added”.
The results are shown in Table 3.

  As shown in Table 3, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent TNF-α production inhibitory action.

[Test Example 4] Hyaluronidase activity inhibitory action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for hyaluronidase activity inhibitory action as follows.

  A hyaluronidase solution (Type IV-S (from bovine testis)) was added to 0.2 mL of 0.1 mol / L acetate buffer (pH 3.5) in which the sample (samples 1 to 3) was dissolved (see Table 4 below for the sample concentration). , SIGMA, 400 NFunits / mL) 0.1 mL was added and reacted at 37 ° C. for 20 minutes. Furthermore, 0.2 mL of 2.5 mmol / L calcium chloride was added as an activator and reacted at 37 ° C. for 20 minutes. To this, 0.5 mL of 0.4 mg / mL potassium hyaluronate solution (from Rooster Comb) was added and reacted at 37 ° C. for 40 minutes.

  Thereafter, 0.2 mL of 0.4 mol / L sodium hydroxide was added to stop the reaction, and after cooling, 0.2 mL of boric acid solution was added to each reaction solution and boiled for 3 minutes. After cooling with ice, 6 mL of p-DABA reagent (p-Dimethylaminobenzaldehyde, manufactured by Wako Pure Chemical Industries, Ltd.) was added and reacted at 37 ° C. for 20 minutes. Thereafter, the absorbance at a wavelength of 585 nm was measured. A blank test was performed and corrected in the same manner. From the obtained measurement results, the hyaluronidase activity inhibition rate (%) was calculated by the following formula.

Hyaluronidase activity inhibition rate (%) = {1− (A−B) / (C−D)} × 100
In the formula, A represents “absorbance at a wavelength of 585 nm when a sample is added / enzyme solution added”, B represents “absorbance at a wavelength of 585 nm when a sample is added / no enzyme solution is added”, and C is “no sample added / enzyme” D represents the “absorbance at a wavelength of 585 nm when no sample was added and no enzyme solution was added”.
The results are shown in Table 4.

  As shown in Table 4, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent hyaluronidase activity inhibitory action.

[Test Example 5] Hexosaminidase release inhibitory action test The loquat flower part extracts (Samples 1 to 3) obtained in Production Examples 1 to 3 were 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 are diluted to a cell density of 4.0 × 10 ⁵ cells / mL, DNP-specific IgE is added to a final concentration of 0.5 μg / 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 washed twice with 500 μL of Siligalian buffer. Next, 30 μL of the same buffer solution and 10 μL of a sample solution prepared with the same buffer solution (samples 1 to 3, see the following Table 5 for the sample concentration) were added, and the mixture was allowed to stand at 37 ° C. 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 cell supernatant in each well and 10 μL of 1 mmol / L p-NAG (p-nitrophenyl-N-acetyl-β-D-glucoside) solution were added to a new 96-well plate and incubated at 37 ° C. for 1 hour. Reacted.

After completion of the reaction, 250 μL of 0.1 mol / L Na ₂ CO ₃ / NaHCO ₃ was added to each well, and the absorbance at a wavelength of 415 nm was measured. As a blank test, the absorbance at a wavelength of 415 nm of a mixed solution of 10 μL of cell supernatant and 250 μL of 0.1 mol / L Na ₂ CO ₃ / NaHCO ₃ was measured and corrected. 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 “absorbance at a wavelength of 415 nm without addition of a sample”, B represents “absorbance at a wavelength of 415 nm with addition of a sample”, and C represents “at a wavelength of 415 nm with addition of a sample and no p-NAG added”. Absorbance ".
The results are shown in Table 5.

  From the results shown in Table 5, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent hexosaminidase release inhibitory action.

[Test Example 6] Tyrosinase activity inhibitory action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for tyrosinase activity inhibitory action as follows.

  In a 48-well plate, McClvaine buffer solution (pH 6.8) 0.2 mL, 0.3 mg / mL tyrosine solution 0.06 mL, 25% DMSO solution of sample (samples 1 to 3, sample concentration shown in Table 6 below) 0.18 mL was added and allowed to stand at 37 ° C. for 10 minutes. To this, 0.02 mL of 800 units / mL tyrosinase solution was added, followed by reaction at 37 ° C. for 15 minutes. After completion of the reaction, the absorbance at a wavelength of 475 nm was measured. A blank test was conducted in the same manner. From the obtained measurement results, the tyrosinase activity inhibition rate (%) was calculated by the following formula.

Tyrosinase activity inhibition rate (%) = {1− (St−Sb) / (Ct−Cb)} × 100
In the formula, St is “absorbance when enzyme solution is added / sample solution is added”, Sb is “absorbance when enzyme solution is not added / sample solution is added”, and Ct is “absorbance when enzyme solution is added / sample solution is not added”. Cb indicates “absorbance when no enzyme solution is added / sample solution is not added”.
The results are shown in Table 6.

  As shown in Table 6, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent tyrosinase activity inhibitory action.

[Test Example 7] Melanin production inhibitory action test The loquat flower part extracts (Samples 1 to 3) obtained in Production Examples 1 to 3 were tested for melanin production inhibitory action as follows.

B16 melanoma cells were cultured using Dulbecco MEM containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco MEM containing 10% FBS and 1 mmol / L theophylline to a cell density of 25.0 × 10 ⁴ cells / mL, then seeded on a 48-well plate at 300 μL per well and cultured for 6 hours.

  After completion of the culture, 300 μL of a sample solution dissolved in Dulbecco's MEM containing 10% FBS and 1 mmol / L theophylline (Samples 1 to 3; see Table 7 below for the sample concentration) was added to each well and cultured for 4 days. After completion of the culture, the medium was removed from each well, 200 μL of 2 mol / L NaOH solution was added, the cells were disrupted with an ultrasonic crusher, and the absorbance at a wavelength of 475 nm was measured to obtain the amount of melanin produced.

  Moreover, in order to evaluate the melanin production inhibitory action per unit cell, after culture | cultivating similarly to the above, the culture solution was removed, and neutral red dissolved in Dulbecco's MEM containing 10% FBS at a final concentration of 0.05 mg / mL Was added to each well and cultured for 2.5 hours. After incubation, the neutral red solution was discarded, and 200 μL of ethanol / acetic acid solution (ethanol: acetic acid: water = 50: 1: 49) was added to each well to extract the dye. After extraction, the absorbance at a wavelength of 540 nm was measured.

  Furthermore, as a blank test, the absorbance at a wavelength of 475 nm and the absorbance at 540 nm were measured for cells cultured in the same manner as described above without adding a sample. From the obtained results, the melanin production inhibition rate (%) per unit cell was calculated by the following formula.

Melanin production inhibition rate (%) = {1− (B / D) / (A / C)} × 100
In the formula, A represents “absorbance at 475 nm when no sample is added”, B represents “absorbance at 475 nm when no sample is added”, C represents “absorbance at 540 nm when no sample is added”, and D represents “ The absorbance at 540 nm at the time of sample addition is expressed.
The results are shown in Table 7.

  As shown in Table 7, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent melanin production inhibitory action.

[Test Example 8] Endothelin-1 mRNA expression increase inhibitory action test With respect to loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3, the endothelin-1 mRNA expression increase suppressive action was tested as follows. .

Normal human epidermis keratinocyte (NHEK) in normal human epidermis keratinocyte growth medium (EpiLife-KG2) in an 80 cm ² flask at 37 ° C., 5% CO ₂ -95% air The cells were precultured under the above conditions and cells were collected by trypsin treatment.

EpiLife-KG2 was used to seed 40 × 10 ⁴ cells / 2 mL / dish in 35 mm dishes (FALCON) and cultured overnight at 37 ° C. and 5% CO ₂ -95% air. After 24 hours, the culture solution was discarded, 1 mL of HEPES buffer was added, UV-B irradiation (50 mJ / cm ² ) was performed, and then the test samples dissolved in EpiLife-KG2 to the required concentrations (samples 1 to 3, sample concentrations are as follows) 2 mL was added to each petri dish, and the cells were cultured for 24 hours under conditions of 37 ° C. and 5% CO ₂ -95% air. After culturing, the culture solution is discarded, 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 levels of mRNA for endothelin-1 and GAPDH as an internal standard were measured. 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 endothelin-1 mRNA is the value of GAPDH 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. Then, the correction values were calculated, and the correction values for UV irradiation / no sample addition and UV irradiation / sample addition when the correction value for UV non-irradiation / no sample addition was 100 were calculated. From the obtained results, the endothelin-1 mRNA expression inhibition rate (%) was calculated by the following formula.

mRNA expression 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 8.

  As shown in Table 8, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent endothelin-1 mRNA expression increase suppressing action.

[Test Example 9] SCF mRNA expression increase inhibitory action test The loquat flower part extracts (Samples 1 to 3) obtained in Production Examples 1 to 3 were tested for SCF mRNA expression increase suppressive action as follows.

Normal human epidermis keratinocyte (NHEK) in normal human epidermis keratinocyte growth medium (EpiLife-KG2) in an 80 cm ² flask at 37 ° C., 5% CO ₂ -95% air The cells were precultured under the above conditions and cells were collected by trypsin treatment.

EpiLife-KG2 was used to seed 40 × 10 ⁴ cells / 2 mL / dish in 35 mm dishes (FALCON) and cultured overnight at 37 ° C. and 5% CO ₂ -95% air. After 24 hours, the culture solution was discarded, 1 mL of HEPES buffer was added, UV-B irradiation (50 mJ / cm ² ) was performed, and then the test samples dissolved in EpiLife-KG2 to the required concentrations (samples 1 to 3, sample concentrations are as follows) 2 mL was added to each petri dish, and cultured under conditions of 37 ° C., 5% CO ₂ -95% air for 24 hours. After culturing, the culture solution is discarded, 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. 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 endothelin-1 mRNA is the value of GAPDH 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. Then, the correction values were calculated, and the correction values for UV irradiation / no sample addition and UV irradiation / sample addition when the correction value for UV non-irradiation / no sample addition was 100 were calculated. From the obtained results, the SCF mRNA expression inhibition rate (%) was calculated by the following formula.

mRNA expression 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 9.

  As shown in Table 9, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent SCF mRNA expression increase inhibitory action.

[Test Example 10] POMC mRNA expression increase suppression action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for POMC mRNA expression increase suppression action as follows.

Normal human epidermis keratinocyte (NHEK) in normal human epidermis keratinocyte growth medium (EpiLife-KG2) in an 80 cm ² flask at 37 ° C., 5% CO ₂ -95% air The cells were precultured under the above conditions and cells were collected by trypsin treatment.

EpiLife-KG2 was used to seed 40 × 10 ⁴ cells / 2 mL / dish in 35 mm dishes (FALCON) and cultured overnight at 37 ° C. and 5% CO ₂ -95% air. After 24 hours, the culture solution was discarded, 1 mL of HEPES buffer was added, UV-B irradiation (50 mJ / cm ² ) was performed, and then the test samples dissolved in EpiLife-KG2 to the required concentrations (samples 1 to 3, sample concentrations are as follows) 2 mL was added to each petri dish, and cultured at 37 ° C. and 5% CO ₂ -95% air for 24 hours. After culturing, the culture solution is discarded, 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 POMC and the internal standard GAPDH mRNA was measured. 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 endothelin-1 mRNA is the value of GAPDH 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. Then, the correction values were calculated, and the correction values for UV irradiation / no sample addition and UV irradiation / sample addition when the correction value for UV non-irradiation / no sample addition was 100 were calculated. From the obtained results, the POMC mRNA expression inhibition rate (%) was calculated by the following formula.

mRNA expression 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 10.

  As shown in Table 10, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent POMC mRNA expression increase inhibitory action.

[Test Example 11] MMP-1 activity inhibitory action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for MMP-1 activity inhibitory action as follows.

  Sample solution dissolved in 0.1 mol / L Tris-HCL buffer (pH 7.1) containing 20 mmol / L calcium chloride in a test tube with a lid (samples 1 to 3, see Table 11 below for sample concentration) ) 50 μL, MMP-1 solution (COLLAGENASE Type IV from Clostridium histolyticum, Sigma) 50 μL and Pz-peptide solution (Pz-Pro-Leu-Gly-Pro-D-Arg-OH, BACHEM Feinchemikalien AG) 400 μL After mixing and reacting at 37 ° C. for 30 minutes, 1 mL of a 25 mmol / L citric acid solution was added to stop the reaction.

  Thereafter, 5 mL of ethyl acetate was added and shaken vigorously. This was centrifuged (1600 × g, 10 minutes), and the absorbance of the ethyl acetate layer at a wavelength of 320 nm was measured. Similarly, a blank test was performed and corrected. From the obtained results, the MMP-1 activity inhibition rate (%) was calculated by the following formula.

MMP-1 activity inhibition rate (%) = {1− (C−D) / (A−B)} × 100
In the formula, A represents “absorbance at a wavelength of 320 nm when no sample is added and an enzyme is added”, B represents “absorbance at a wavelength of 320 nm when no sample is added and an enzyme is not added”, and C is “addition of sample and enzyme is added” And “D” represents “absorbance at a wavelength of 320 nm when a sample is added and no enzyme is added”.
The results are shown in Table 11.

  As shown in Table 11, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent MMP-1 activity inhibitory action.

[Test Example 12] Type I collagen production promoting action test The loquat flower part extracts (Samples 1 to 3) obtained in Production Examples 1 to 3 were tested for type I collagen production promoting action as follows.

Human normal fibroblasts (Detroit 551) were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsinization. The collected cells were diluted with the above medium to a cell density of 1.6 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well microplate and cultured overnight. After completion of the culture, the medium was removed, and 150 μL of a sample solution (samples 1 to 3, sample concentration shown in Table 12 below) dissolved in 0.25% FBS-containing Dulbecco MEM medium was added to each well for 3 days. Cultured. After completion of the culture, the amount of type I collagen in the medium of each well was measured by ELISA. Similarly, the amount of type I collagen was measured without adding the sample solution. From the obtained results, the type I collagen production promotion rate (%) when the sample solution was added was calculated according to the following formula.

Type I collagen production promotion rate (%) = A / B × 100
In the formula, A indicates “the amount of type I collagen when the sample solution is added”, and B indicates “the amount of type I collagen when no sample solution is added”.
The results are shown in Table 12.

  As shown in Table 12, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent type I collagen production promoting action.

[Test Example 13] Type IV collagen production promoting action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for type IV collagen production promoting action as follows.

Human normal fibroblasts (Detroit 551) were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsinization. The collected cells were diluted with the above medium to a cell density of 1.6 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well microplate and cultured overnight. After completion of the culture, the medium was removed, and 150 μL of a sample solution (samples 1 to 3, sample concentration shown in Table 13 below) dissolved in 0.25% FBS-containing Dulbecco MEM medium was added to each well for 3 days. Cultured. After completion of the culture, the amount of type IV collagen in the medium of each well was measured by ELISA. Similarly, the amount of type IV collagen was measured without adding the sample solution. 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 the sample solution is added”, and B indicates “amount of type IV collagen when no sample solution is added”.
The results are shown in Table 13.

  As shown in Table 13, the loquat flower part extract (samples 1 to 3) was confirmed to have an excellent type IV collagen production promoting action.

[Test Example 14] Damage recovery action test by ultraviolet (UV-B) irradiation About loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3, ultraviolet (UV-B) was as follows. The damage recovery effect by irradiation was tested.

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 α-MEM medium to a cell density of 2.0 × 10 ⁵ cells / mL, and then seeded at 200 μL per well in a 48-well plate. After culturing for 24 hours, the medium was replaced with 100 μL of PBS (−) and irradiated with 1.0 J / cm ² of UV-B. Immediately after irradiation, PBS (-) was removed, and 400 μL of sample solution dissolved in D-MEM containing 10% FBS (Samples 1 to 3, sample concentration shown in Table 14 below) was added to each well for 24 hours. Cultured.

  The damage recovery effect by ultraviolet (UV-B) irradiation was measured using an MTT assay. After completion of the culture, the medium was removed, and 200 μL of MTT dissolved at a final concentration of 0.4 mg / mL was added to each well. After culturing for 2 hours, blue formazan produced in the cells was extracted with 200 μL of 2-propanol, and after extraction, absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. Similarly, after seeding the cells, the cells not irradiated with UV-B and the cells irradiated with UV-B but not added with the sample solution were also measured in the same manner, and were set as a non-irradiated group and an irradiated group, respectively. From the obtained results, the damage recovery rate (%) by ultraviolet (UV-B) irradiation was calculated by the following formula.

Damage recovery rate (%) = {(Nt−C) − (Nt−Sa)} / (Nt−C) × 100
In the formula, Nt represents “absorbance in cells not irradiated with UV-B”, C represents “absorbance in cells irradiated with UV-B but not added with a sample solution”, and Sa represents “UV -Absorbance in cells irradiated with B and added with sample solution ".
The results are shown in Table 14.

  As shown in Table 14, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent damage recovery effect by ultraviolet (UV-B) irradiation.

[Test Example 15] Cyclic AMP phosphodiesterase activity inhibitory action test The loquat flower part extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for cyclic AMP phosphodiesterase activity inhibitory action as follows. .

  0.1 mL of a 2.5 mg / mL bovine serum albumin solution, 0.1 mg / mL cyclic AMP phosphodiesterase solution in 0.2 mL of 50 mM Tris-HCl buffer (pH 7.5) containing 5 mM magnesium chloride 0.05 mL of a sample solution dissolved in 1 mL and 50% DMSO (Samples 1 to 3, see sample table below for sample concentration) was added, and incubated at 37 ° C. for 5 minutes.

  To this reaction solution, 0.05 mL of a 0.5 mg / mL cyclic AMP solution was added and incubated at 37 ° C. for 60 minutes. The reaction was stopped by boiling on a boiling water bath for 3 minutes, this was centrifuged (2260 × g, 10 minutes, 4 ° C.), and cyclic AMP, which is a reaction substrate in the supernatant, was subjected to a high-speed liquid under the following conditions. Analyzed using chromatography. In addition, a blank test was performed in the same manner without adding the sample solution.

<High performance liquid chromatography conditions>
Product name: Chromatocorder 12 (manufactured by SYSTEM INSTRUMENTS)
Stationary phase: Wakosil 5C ₁₈ (Wako Pure Chemical Industries, Ltd.)
Column diameter: 4.6 mm
Column length: 250mm
Mobile phase: 1 mM TBAP in 25 mM KH ₂ PO ₄ : CH ₃ CN = 90: 10
Mobile phase flow rate: 1.0 mL / min
Detection: UV, 260nm

  Next, the peak area (A) of the cyclic AMP standard product, the peak area (B1) of the supernatant of the reaction solution of the cyclic AMP standard product and cyclic AMP phosphodiesterase when no sample is added, and the cyclic when the sample is added The peak area (B2) of the supernatant of the reaction solution of AMP standard product and cyclic AMP phosphodiesterase was determined. From the obtained results, the decomposition rate (C) of the cyclic AMP standard product when no sample was added and the decomposition rate (D) of the cyclic AMP standard product when the sample was added were calculated according to the following equations.

Standard product decomposition rate when no sample is added (C,%) = (1−B1 / A) × 100
Decomposition rate of standard product at the time of sample addition (D,%) = (1−B2 / A) × 100

Thereafter, the cyclic AMP phosphodiesterase activity inhibition rate (%) was calculated by the following formula based on the respective degradation rates (C, D) calculated by the above formula.
Inhibition rate (%) = (1-D / C) × 100
The results are shown in Table 15.

  As shown in Table 15, it was confirmed that the loquat flower part extract (samples 1 to 3) has an excellent cyclic AMP phosphodiesterase activity inhibitory action.

[Formulation Example 1]
An emulsion having the following composition was produced by a conventional method.
Biwa flower part extract (any one of Production Examples 1 to 3) 0.1 g
Jojoba oil 4.0g
Olive oil 2.0g
Squalane 2.0g
Cetanol 2.0g
Glyceryl monostearate 2.0g
Polyoxyethylene cetyl ether (20E.O.) 2.5g
Oleic acid polyoxyethylene sorbitan (20E.O.) 2.0g
Twilight extract 0.1g
0.1g dipotassium glycyrrhizinate
Ginkgo biloba extract 0.1g
Conchiolin 0.1g
Oat extract 0.1g
Chamomile extract 0.1g
1,3-butylene glycol 3.0 g
Methyl paraoxybenzoate 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.
Biwa flower part extract (any one of Production Examples 1 to 3) 0.1 g
Glycerin 3.0g
1,3-butylene glycol 3.0 g
Oleic acid polyoxyethylene sorbitan (20E.O.) 0.5g
Methyl paraoxybenzoate 0.15g
Citric acid 0.1g
Sodium citrate 0.1g
Oil soluble licorice extract 0.1g
Seaweed extract 0.1g
Xylobiose Mixture 0.5g
Kujin extract 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.
Biwa flower part extract (any one of Production Examples 1 to 3) 1.1 g
Liquid paraffin 5.0g
Salami beeswax 4.0g
Cetanol 3.0g
Squalane 10.0g
Lanolin 2.0g
Stearic acid 1.0g
Oleic acid polyoxyethylene sorbitan (20E.O.) 1.5g
3.0 g glyceryl monostearate
1,3-butylene glycol 6.0 g
Fragrance 0.1g
Purified water remainder (total amount is 100 g)

[Formulation Example 4]
The following mixture was tableted to produce a tablet-shaped dietary supplement.
Loquat flower part extract (any one of Production Examples 1 to 3) 50 parts by weight Powdered sugar (sucrose) 188 parts by weight Glycerin fatty acid ester 12 parts by weight

[Formulation Example 5]
The following mixture was granulated to produce a dietary supplement.
Loquat flower part extract (any one of Production Examples 1 to 3) 50 parts by weight Beet oligosaccharide 1000 parts by weight Vitamin C 167 parts by weight Stevia extract 10 parts by weight

  The antioxidant of the present invention is used to prevent and improve various disorders caused by reactive oxygen species, the anti-inflammatory agent of the present invention is used to prevent and improve various inflammatory diseases, and the whitening agent of the present invention is stains and buckwheat. The anti-aging agent of the present invention is useful for the prevention and improvement of skin aging, the anti-obesity agent or the cyclic AMP phosphodiesterase activity inhibitor of the present invention is used for the prevention and improvement of skin pigmentation, etc.・ Contribute greatly to improvement.

Claims (7)

  A whitening agent comprising an extract from loquat flower as an active ingredient. The extract from the flower part of the loquat is a tyrosinase activity inhibitory action, a melanin production inhibitory action, an endothelin-1 mRNA expression rise inhibitory action, a stem cell growth factor (SCF) mRNA expression rise inhibitory action, and a proopiomelanocortin (POMC) mRNA expression rise. The whitening agent according to claim 1 , wherein the whitening agent has one or more actions selected from the group consisting of inhibitory actions.   An anti-aging agent comprising an extract from loquat flower as an active ingredient. The extract from the flower part of the loquat is selected from the group consisting of matrix metalloproteinase-1 (MMP-1) activity inhibitory action, type I collagen production promoting action, type IV collagen production promoting action and damage recovery action by ultraviolet irradiation The anti-aging agent according to claim 3 , which has one or more actions.   An anti-obesity agent comprising an extract from loquat flower as an active ingredient.   A cyclic AMP phosphodiesterase activity inhibitor comprising an extract from loquat flower as an active ingredient. A skin cosmetic comprising an extract from a flower part of loquat and used for one or more uses selected from the group consisting of whitening, anti-aging and anti-obesity .