JP2012116765A - Cgrp responsiveness promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a material for promoting CGRP (calcitonin gene-related peptide) responsiveness of cells.SOLUTION: There is provided the CGRP responsiveness promoter of cells containing at least one substance selected from the group consisting of butcher's broom, Amur corktree, Hemerocallis fulva, rose balsam, fucoid, birch, rice protein, rice oil, Aloe vera, root sheath of Paeonia moutan, laminaria, star fruit, cornflower, lemon grass, apricot kernel, lemon, sweet marjoram, marronnier, royal jelly, rose hip and their extracts as an active component.

Description

  The present invention relates to a CGRP (calcitonin gene-related peptide) response promoter.

  In the skin, a vascular network composed of three layers of vascular beds parallel to the skin surface is formed. The primary function of skin circulation is body temperature regulation. When cold or thermal stimulation is applied, the skin blood vessels contract or expand or relax to change the blood flow of the skin circulation, thereby suppressing or promoting body heat dissipation. To maintain body temperature. Skin circulation also plays an important role in regulating the distribution of blood flow throughout the body and is caused by information and movement from pressure, volume or chemoreceptors, as well as information from central and peripheral thermal receptors. Also affected by vasomotor reflexes.

  Skin circulation undergoes changes with age. It is known that the structure of the skin vascular network and the blood flow volume change with aging, and the width of the blood flow change with respect to cold stimulation and thermal stimulation is reduced. As a result, there may be a problem that the blood circulation of the skin deteriorates with aging, thereby further reducing the metabolism of the skin.

  Skin circulation is subject to dual regulation, a systemic mechanism via the sympathetic nervous system and a local mechanism by local regulators. One locality mechanism is axon reflection. This is because impulses generated in the sensory nerves of the skin by stimulation are transmitted retrogradely to other branches in the middle of conduction to the center, releasing neuropeptides from the nerve endings and dilating skin blood vessels It is a phenomenon. The neuropeptide released at this time is substance P (SP) or calitonin gene-related peptide (CGRP).

  CGRP is a 37-amino acid neuropeptide that is biosynthesized by tissue-specific alternative splicing of the calcitonin gene. CGRP is widely distributed in the central nervous system and peripheral sensory nervous system, and exerts its action through specific receptors. The relationship between CGRP and blood flow is well known, and skin blood flow is increased by administration of CGRP in rats (Non-patent document 1), blood pressure of CGRP knockout mice is increased (Non-patent document 2), Rats administered with CGRP neutralizing antibody have reduced skin blood flow (Non-patent Document 3), and patients with Raynaud's disease, which are thought to be caused by excessive contraction of peripheral blood vessels, lack CGRP-containing nerves in the fingers. It is known that non-patent document 4 and the skin blood flow of the arm is increased by injecting CGRP intravenously to a Raynaud disease patient (non-patent document 5).

In addition to vasodilatory effects, CGRP has physiological effects such as regulation of substance P in inflammation, vascular permeability enhancement, regulation of nicotinic receptors at the neuromuscular junction, inhibition of gluconeogenesis and promotion of glycolysis, pancreatic enzyme Secretion promotion, gastric acid secretion suppression, heart rate promotion, nerve activity regulation, calcium metabolism regulation, bone formation promotion, insulin secretion, body temperature rise, food intake decline. It also has anti-inflammatory effects through prostaglandin synthesis, suppression of H ₂ O ₂ production and antigen presentation in macrophages, dentinal blast cell division, preconditioning for ischemia-reperfusion injury, and myometrium and uterus. Involvement in pregnancy and parturition is known because the receptor is expressed in the placenta.

  Because of the physiological effects described above, CGRP is used for the treatment of blood circulation, wound healing, and hypertension, heart failure, liver damage caused by ischemic reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage Pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor, and the like. Further, the use of CGRP receptor antagonists includes treatment of migraine, hypersensitivity, and the like. In fact, many CGRP receptor antagonist compounds have been developed as anti-migraine agents.

  A substance capable of promoting or suppressing the responsiveness of skin circulation to CGRP is useful for improving the change in skin circulation due to aging, and for preventing or treating the above diseases and symptoms. So far, as a substance capable of suppressing CGRP responsiveness, a CGRP receptor antagonist, an anti-CGRP antibody, and an iris extract (Patent Document 1) are known. However, it is desired to develop further substances that can promote or suppress CGRP responsiveness and are useful for regulating skin circulation or improving various diseases.

Patent No. 3093154

Nature, 1988, 335: 164-167 Hypertension, 2000, 35: 470-475 Br J Pharmacol, 2008, 155: 1093-1103 Lancet, 1990, 336: 1530-1533 Lancet, 1989, 2 (8676): 1354-1357

  The present invention relates to the provision of a CGRP responsive promoter for cells.

  The present inventors searched for a material that can promote CGRP responsiveness of cells.

That is, the present invention provides the following.
(1) Butcher Bloom, Yellowfin, Hemerocallis Fulva, Buttercup, Hibamata, Birch, Rice Protein, Rice Oil, Aloe Vera, Buttonpi, Laminaria, Star Fruit, Cornflower, Lemongrass, Kyonin, Lemon, Sweet Marjoram, Maronier, Royal Jelly, Rosehip And a CGRP responsive promoter for cells, comprising as an active ingredient at least one of materials selected from the group consisting of these and extracts thereof.
(2) The promoter according to (1), wherein the cell is a vascular smooth muscle cell.
(3) A method for promoting CGRP responsiveness of a cell, the cell having a CGRP receptor and wishing to promote CGRP responsiveness, butcher bloom, yellowfin, hemerocallis fluva, spinach, hibermata, birch, rice protein, Existence of at least one material selected from the group consisting of rice oil, aloe vera, buttonpi, laminaria, star fruit, cornflower, lemongrass, kyounin, lemon, sweet marjoram, maronier, royal jelly, rosehip and extracts thereof A method comprising the step of culturing below.
(4) The method according to (3), wherein the cell is a vascular smooth muscle cell.

  ADVANTAGE OF THE INVENTION According to this invention, the CGRP responsiveness promoter which can promote CGRP responsiveness of a cell is provided. The CGRP response promoter is useful for improving changes in skin circulation due to aging and the like, and for preventing or treating various diseases and symptoms related to CGRP.

  In the present specification, “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human body by therapy.

  In the present specification, “improvement” refers to improvement of a disease or symptom, prevention or delay of worsening of the disease or symptom, or reversal, prevention or delay of progression of the disease or symptom.

  As used herein, “prevention” refers to preventing or delaying the onset of a disease or symptom in an individual, or reducing the risk of developing an individual's disease or symptom.

In the present specification, “promoting CGRP responsiveness of cells” refers to promoting cell activity induced by CGRP. When CGRP binds to a cellular CGRP receptor, an increase in intracellular cAMP occurs through activation of adenylate cyclase via the G protein. Thereby, for example, in vascular smooth muscle cells, K ⁺ channels are opened through protein kinase A activation. Alternatively, in vascular endothelial cells, eNOS is activated through protein kinase A activation, and NO production is promoted. Furthermore, the produced NO acts on vascular smooth muscle cells, and K ⁺ channels are opened through intracellular cGMP activation via NO.
The cellular CGRP responsive promoters according to the present invention may promote a series of these processes in cells triggered by CGRP. For example, the action of the CGRP responsive promoter of the present invention includes promotion of binding between CGRP and its receptor, enhancement of G protein activity, enhancement of adenylate cyclase activity, increase of intracellular cAMP, protein kinase A activity Enhancement, NO production or cGMP activity enhancement, and K ⁺ channel opening promotion. Preferably, the “cell CGRP responsiveness” can be determined using the degree of increase in intracellular cAMP caused by CGRP as an index. Therefore, the CGRP responsive promoter of the present invention preferably promotes an increase in intracellular cAMP induced by CGRP.

In the present invention, the “cell” that promotes CGRP responsiveness is not particularly limited as long as it is a cell that expresses a CGRP receptor or a cell that has a CGRP receptor. Preferably, the cells include vascular smooth muscle cells, vascular endothelial cells, fibroblasts, osteoblasts, enamel blasts, dentin blasts, and more preferably vascular smooth muscle cells and vascular endothelial cells. .
Alternatively, the “cell” may be a cell fragment or a cell fraction of the cells listed above, or may be a tissue containing the cells listed above or a culture derived from the cells listed above. Good.

In this specification, “Butcher Bloom” refers to Ruscus aculeatus of the lily family, “Kihada” refers to Phellodendron amurense of the citrus family, “ Hemerocallis fulva ” refers to Hemerocallis fulva of the lily family, `` Hempensis '' refers to the Impatiens balsamina of the scorpionaceae , `` Hibamata '' refers to the Fucus vesiculosus of the Asteraceae , `` Birch '' refers to the birch plant, preferably the European birch Betula pendula , `` Aloe vera ”Refers to the Aloe vera of the aloe family,“ button pi ”refers to the root bark of Paeonia moutan of the button family,“ laminaria ”refers to the Laminaria genus algae,“ star fruit ”refers to the Oxalis family refers to the Averrhoa carambola, is a "cornflower", refers to the centaurea cyanus of the Asteraceae, is a "lemon grass", refers to the Cymbopogon citratus of the grass family, "yellow The Unin ", refers to the seeds of the Rosaceae Prunus plant, a" lemon ", refers to the Citrus limon of the citrus family, the" sweet marjoram ", refers to the Origanum majorana of the mint family, and" horse chestnut "is, Aesculus hippocastanum of the aceae family, and “rose hip” refers to the fruit of the Rosaceae, preferably Rosa canina . In the present invention, “rice” from which “rice protein” and “rice oil” are derived means the seeds of Gramineae, and “royal jelly” means a secretory gland (large gland) in the head of a worker bee. This refers to substances secreted by the urinary glands and hypopharyngeal glands.

  When the above-mentioned material is a plant, the extract is not particularly limited, and any part of the plant, for example, whole grass, leaves, stems, buds, flowers, buds, wood parts, bark, lichens, roots, What is necessary is just an extract from a rhizome, a pseudo corm, a corm, a tuber, a seed, a fruit, a mycorrhiza, a resin, etc., or those combinations. Preferred sites for obtaining the extract are rhizomes for butcher bloom, bark for yellowfin, flowers for Hemerocallis fulva, flowers or aerial parts for spinach, algae for hibermata, for birch Bark, leaves for aloe vera, root bark for buttonpi, whole plant for laminaria, leaves for star fruit, flowers for cornflower, leaves for lemongrass, seeds for ginger, lemon In the case of sweet, the leaves in the case of sweet marjoram, the fruit in the case of maronier, and the fruit in the case of rosehip. The parts listed above may be subjected to the extraction step as they are, or may be subjected to the extraction step after being pulverized, cut or dried.

  As the above extract, commercially available ones may be used, or various solvent extracts obtained by a conventional method, or diluted solutions thereof, concentrated solutions thereof, dried powders thereof, pastes or activated carbon treatments thereof. There may be. As an example, the extract in the present invention can be obtained by extracting these plants at room temperature or under heating, or by using an extraction device such as a Soxhlet extractor.

  As the solvent for extraction, either a polar solvent or a nonpolar solvent can be used. Specific examples of the solvent include water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; methyl acetate and ethyl acetate Esters; linear and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as squalane, hexane, cyclohexane and petroleum ether; aromatic hydrocarbons such as toluene; dichloromethane and chloroform And halogenated hydrocarbons such as dichloroethane; and supercritical carbon dioxide; pyridines; organic solvents such as oils and fats, other oils such as wax; and mixtures thereof. Preferable examples include water, alcohols and aqueous solutions thereof, and ethanol is preferable as the alcohols. More preferred solvents are water and an aqueous ethanol solution.

As a compounding ratio (volume ratio) of alcohol and water, 0.001-100: 99.999-0 are preferable, 5-95: 95-5 are more preferable, 20-80: 80-20 are more preferable. 30-70: 70-30 is still more preferable, and 40-60: 60-40 is still more preferable. In the case of an ethanol aqueous solution, the ethanol concentration is preferably 40 to 60% by volume.
As a usage-amount of a solvent, 1-100 mL is preferable with respect to 1 g of raw materials (dry mass conversion). The extraction conditions are not particularly limited as long as extraction is performed for a time that allows sufficient extraction at an extraction temperature between 0 ° C. and the boiling point of the solvent. Usually, extraction is performed for a long time at a low temperature and for a short time at a high temperature.
Specific examples of the extraction means for obtaining the extract include solid-liquid extraction, liquid-liquid extraction, immersion, decoction, leaching, reflux extraction, ultrasonic extraction, microwave extraction, and stirring.
Examples of extraction conditions include 15 to 25 ° C. for 7 to 14 days, about 70 ° C. for 5 hours, and the like.
Moreover, when shortening extraction time, solid-liquid extraction with stirring is desirable. An example of suitable conditions for this solid-liquid extraction is stirring at 40 to 100 ° C. (preferably 50 to 70 ° C.) at 100 to 1000 rpm for 30 to 300 minutes.
In order to prevent the oxidation of the extract, a means for extracting under a so-called non-oxidizing atmosphere while removing dissolved oxygen by bubbling degassing or inert gas such as nitrogen gas may be used in combination.

  As shown in the examples below, butcher bloom, yellowfin, hemerocallis fluva, spinach, hibermata, birch, rice protein, rice oil, aloe vera, buttonpi, laminaria, star fruit, cornflower, lemongrass, kyounin, lemon, sweet marjoram, maronier In addition, a material selected from the group consisting of royal jelly, rosehip and extracts thereof has a function of significantly promoting CGRP responsiveness of cells. Therefore, these materials are useful as cell CGRP responsiveness promoters. In addition, these materials can exert an effect of improving skin circulation through the CGRP responsiveness promoting action, and as a result, skin metabolism improvement, blood circulation promotion, wound healing promotion, high blood pressure, heart failure, false Prevention and / or prevention of diseases such as liver damage due to blood reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor Or it is useful for improvement.

  That is, the above-mentioned material is used for promoting CGRP responsiveness of cells, for improving skin circulation or skin metabolism, for promoting blood circulation, for promoting wound healing, or for liver damage caused by hypertension, heart failure, ischemic reperfusion. , Renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, premature labor, etc. can do. The use can be in humans or non-human animals, or tissues, organs, cells derived therefrom, and can be therapeutic or non-therapeutic. Each of the above materials may be used alone or in any combination of two or more.

  Accordingly, as one aspect, the present invention is a butcher bloom, yellowfin, hemerocallis fulva, spinach, hibermata, birch, rice protein, rice oil, aloe vera, buttonpi, laminaria, star fruit, cornflower, lemongrass, kyonin, lemon, sweet marjoram And a CGRP responsive promoter for cells containing, as an active ingredient, at least one material selected from the group consisting of Maronnier, Royal Jelly, Rosehip and extracts thereof. The CGRP responsive promoter is used for promoting CGRP responsiveness of cells, improving skin circulation or skin metabolism, promoting blood circulation, promoting wound healing, or by hypertension, heart failure, ischemic reperfusion To prevent and / or improve diseases such as liver damage, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor Can be used for In one embodiment, the CGRP responsiveness promoter of the present invention consists essentially of at least one of the above materials.

  As another aspect, the present invention relates to butcher bloom, yellowfin, hemerocallis fulva, spinach, hibermata, birch, rice protein, rice oil, aloe vera, botpi, laminaria, star fruit, cornflower, lemongrass, kyounin, lemon, sweet marjoram, Provided is a skin circulation improving agent containing, as an active ingredient, at least one of materials selected from the group consisting of maronier, royal jelly, rosehip and extracts thereof. As another aspect, the present invention provides a skin metabolism improving agent containing at least one of the above materials as an active ingredient. In one embodiment, these agents consist essentially of at least one of the above materials. Preferably, the agent for improving skin metabolism is an agent for improving reduction in skin metabolism due to aging or the like.

  As another aspect, the present invention relates to butcher bloom, yellowfin, hemerocallis fulba, spinach, hibermata, birch, rice protein, rice oil, aloe vera, buttonpi, laminaria, star fruit, cornflower, lemongrass, kyonin, lemon, sweet marjoram There is provided a blood circulation promoter containing, as an active ingredient, at least one material selected from the group consisting of Maronnier, Royal Jelly, Rosehip, and extracts thereof. As another aspect, the present invention provides a wound healing promoter containing at least one of the above materials as an active ingredient. As yet another aspect, the present invention relates to hypertension, heart failure, ischemic reperfusion-induced liver damage, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoids containing at least one of the above materials as an active ingredient. Provided is a preventive and / or ameliorating agent for diseases such as bleeding, pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia and premature labor. In one embodiment, these agents consist essentially of at least one of the above materials.

  Butcher Bloom, Yellowfin, Hemerocallis Fulva, Buttercup, Hibermata, Birch, Rice Protein, Rice Oil, Aloe Vera, Buttonpi, Laminaria, Star Fruit, Cornflower, Lemongrass, Kyonin, Lemon, Sweet Marjoram, Maronier, Royal Jelly, Rosehip and Their The material selected from the group consisting of extracts is for promoting CGRP responsiveness of cells, for improving skin circulation or skin metabolism, for promoting blood circulation, for promoting wound healing, or for hypertension Diseases such as heart failure, liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, premature labor Of compositions, medicines, quasi-drugs, cosmetics, foods and drinks, etc. It can be used for. Such compositions, pharmaceuticals, quasi drugs, cosmetics, foods and drinks and the like are also within the scope of the present invention.

  The above composition, medicine, quasi drug, cosmetic, food and drink, etc. can be produced or used for human or non-human animals. The above-mentioned materials or the CGRP responsiveness promoter of the present invention is blended in the composition, medicine, quasi-drug, cosmetics, food and drink, etc. To improve or improve skin metabolism, promote blood circulation, promote wound healing, or hypertension, heart failure, liver damage due to ischemic reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, It may be an active ingredient for preventing and / or improving diseases such as pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, premature labor.

  The pharmaceutical or quasi-drug contains at least one of the above-mentioned materials or the CGRP responsiveness promoter of the present invention as an active ingredient. The pharmaceutical or quasi drug can be administered in any dosage form. Administration may be oral or parenteral. Dosage forms for oral administration include, for example, solid dosage forms such as tablets, coated tablets, granules, powders, capsules, and liquid dosage forms such as elixirol, syrups and suspensions, Examples of the dosage form for parenteral administration include injection, infusion, transdermal, transmucosal, nasal, enteral, inhalation, suppository, bolus, patch and the like.

  The medicine or quasi-drug may contain the above-mentioned materials, or the CGRP responsiveness promoter of the present invention, alone or in combination of any two or more, or further pharmaceutically acceptable. You may contain in combination with a support | carrier. Examples of such carriers include excipients, coating agents, binders, extenders, disintegrating agents, lubricants, diluents, osmotic pressure adjusting agents, pH adjusting agents, dispersing agents, emulsifiers, preservatives, and stabilizers. , Antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, lubricants, activity enhancers, anti-inflammatory agents, bactericides, fragrances, flavoring agents, flavoring agents and the like. Moreover, the said pharmaceutical and quasi-drug may contain another active ingredient and pharmacological component, as long as the CGRP responsiveness promotion effect of the said raw material is not lost.

  The above-mentioned medicine or quasi-drug is prepared from the above-mentioned materials or the CGRP responsive promoter of the present invention, or in combination with the above carriers and / or other active ingredients and pharmacological ingredients as necessary. Can be manufactured. The content of the above-mentioned raw materials in the medicine or quasi-drug is preferably 0.000001 to 100% by mass, more preferably 0.000002 to 100% by mass, as a dry weight. More preferably, it is 0.0000 to 100% by mass, even more preferably 0.00001 to 100% by mass, and still more preferably 0.0001 to 100% by mass.

  The said food / beverage products contain the raw material raised above or the CGRP responsiveness promoter of this invention as an active ingredient. The said food / beverage products can be made into the food / beverage products, the functional food, the food for sick people, and the food for specific health which displayed the effect as needed on the concept of CGRP responsiveness promotion.

  As the form of the food, in addition to various foods such as breads, cakes, noodles, confectionery, jelly, frozen food, ice cream, dairy products, soups, edible oil, seasonings and the like, oral administration described above Forms similar to the formulation (tablets, capsules, granules, powders, syrups, etc.), livestock feeds, pet foods, etc. can be mentioned. Examples of beverage forms include fruit juice drinks, carbonated drinks, tea-based drinks, Examples include near water, sports drinks, milk drinks, alcoholic drinks, and soft drinks. Moreover, a drink can be used as the container-packed drink with which the container was filled.

In order to prepare the above-mentioned various forms of food and drink, the above-described materials or the CGRP responsive accelerator of the present invention alone or other food and drink materials, solvents, softeners, oils, emulsifiers, preservatives , Fragrances, stabilizers, colorants, antioxidants, humectants, thickeners and the like can be used in appropriate combinations.
The content of the material raised above in the food or drink is preferably 0.000001 to 100% by mass, more preferably 0.000002 to 75% by mass, as dry weight, and 0.000005 to The content is more preferably 50% by mass, even more preferably 0.00001-30% by mass, and still more preferably 0.0001-20% by mass.

  The cosmetic contains the above-mentioned materials or the CGRP responsiveness promoter of the present invention as an active ingredient. The cosmetic may contain the above-mentioned materials, or the CGRP responsiveness promoter of the present invention, alone or in combination of any two or more, or further in combination with a carrier acceptable as a cosmetic. You may contain. Examples of such carriers include excipients, coating agents, binders, extenders, disintegrating agents, lubricants, diluents, osmotic pressure adjusting agents, pH adjusting agents, dispersing agents, emulsifiers, preservatives, and stabilizers. , Antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, lubricants, activity enhancers, anti-inflammatory agents, bactericides, fragrances, flavoring agents, flavoring agents and the like. In addition, as long as the CGRP responsiveness promoting action of the above materials is not lost, the cosmetics are other active ingredients and cosmetic ingredients such as moisturizers, whitening agents, UV protection agents, cell activators, detergents, keratolytics. Agents, make-up ingredients (for example, makeup bases, foundations, glistening, powder, teak, lipstick, eye makeup, eyebrow, mascara, etc.) may be contained. Examples of the form of the cosmetic include creams, emulsions, lotions, suspensions, gels, powders, packs, sheets, patches, sticks, cakes, and the like, which can be used for cosmetics.

  The cosmetic can be produced from the above-mentioned materials or the CGRP responsive promoter of the present invention or, if necessary, by combining the carrier and / or other active ingredients and cosmetic ingredients according to a conventional method. it can. The content of the material mentioned above in the cosmetic is preferably 0.000001 to 100% by mass, more preferably 0.000002 to 50% by mass, as dry weight, and 0.000005 to 30%. It is more preferable to set it as the mass%, It is still more preferable to set it as 0.00001-20 mass%, It is still more preferable to set it as 0.0001-10 weight%.

  The present invention also provides a method for promoting CGRP responsiveness of cells. The method includes a step of adding at least one of the above-mentioned materials to a cell having a CGRP receptor and in which CGRP responsiveness is to be promoted. When the cell is a cell culture, the concentration of the above-mentioned raw material added is 0.000001 to 2% (w / v), preferably 0.00001 to 1% (w / v) as a dry weight. And more preferably 0.001 to 0.8% (w / v). The timing which adds the raw material mentioned above should just be before CGRP stimulation and / or at the time of CGRP stimulation.

The above-mentioned materials or the CGRP responsive promoter of the present invention is used for promoting CGRP responsiveness, improving skin circulation or improving skin metabolism, promoting blood circulation, promoting wound healing, or hypertension, Diseases such as heart failure, liver damage caused by ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor For prevention and / or amelioration, it can be administered in an effective amount to a subject in need thereof.
In one embodiment, the administration is performed non-therapeutically with the aim of improving skin circulation, improving skin metabolism, promoting blood circulation for health promotion or cosmetic purposes.

Examples of the administration target include animals that need to improve skin circulation or skin metabolism, and preferably animals that need to improve reduction of skin metabolism due to aging and the like. Alternatively, the subjects of administration include animals that need to promote blood circulation or wound healing, as well as hypertension, heart failure, liver damage due to ischemic reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage , Pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, animals suffering from diseases such as premature labor, animals suspected to be, or animals at high risk. The animal is preferably a human or non-human mammal, more preferably a human.
Alternatively, the administration subject can be an animal-derived tissue, organ, cell, or fraction thereof. The tissue, organ, cell, or fraction thereof, is a naturally-derived or biologically or bioengineered tissue, organ, cell, or a CGRP receptor that expresses or has a CGRP receptor, or These fractions.

  The preferred dosage may vary according to the subject's species, weight, sex, age, condition or other factors. The dose, route, interval, and amount and interval of intake can be determined appropriately by those skilled in the art. For example, when administered orally to humans, the dosage is Butcher Bloom, Yellowfin, Hemerocallis Fulva, Buttercup, Hibermata, Birch, Rice Protein, Rice Oil, Aloe Vera, Buttonpi, Laminaria, Star Fruit, Cornflower, Lemongrass, Kyonin, Lemon The dry weight of sweet marjoram, maronnier, royal jelly, rosehip or extracts thereof is preferably 10 μg to 50000 mg / day, more preferably 100 μg to 30000 mg / day, and 1000 μg to 20000 mg / day per adult. More preferably, 5000 μg to 10000 mg / day is still more preferable.

  EXAMPLES Hereinafter, an Example is shown and this invention is demonstrated more concretely.

Example 1
The effect of promoting CGRP responsiveness of cells by various substances was examined.
(1) Method 1. Cell culture Normal human coronary artery smooth muscle cells (HCASMC; Kurabo Corp., cell lot # 625723) growth medium (basic medium HuMedia-SB2 500ml FBS 25ml, hEGF 0.5ml, hFGF-B 0.5ml, insulin 0.5ml, antibacterial Cultivated with a supplement of 0.5 ml of an agent (Kurabo). For passage, 0.025% trypsin / 0.01% EDTA was used.

2. Preparation of cells Normal human coronary artery smooth muscle cells were seeded in a 48-well plate using a growth medium at a density of 4000 cells / cm ² , and the next day differentiation medium (basic medium HuMedia-SB2 500 ml FBS 5 ml, heparin 0.5 ml The antibacterial agent 0.5ml was added; Kurabo Industries Co., Ltd.). The medium was changed every other day and cultured for 7 days. During the last 2 days, the test substances listed in Table 1 were cultured in a differentiation medium containing 0.1% (v / v), and then the cells were subjected to measurement of CGRP responsiveness.

3. Measurement of CGRP responsiveness Cells were phosphodiesterase inhibitor (PDI) IBMX (3-Isobutyl-1-methylxanthine, 500 μM; Sigma) and Ro-20-1724 (100 μM; Wako Pure Chemical Industries, Ltd.), 0.1% (v / v) was washed twice with a basal medium (Humedia-SB2; Kurabo Industries) containing the test substance shown in Table 1, and 200 μl of the same medium was further added and cultured at 37 ° C. for 60 minutes. Inhibitors were infiltrated. The medium was removed and a basal medium containing 0.1% (v / v) of the test substance described in Table 1, 0.5 nM CGRP, and a phosphodiesterase inhibitor was added and incubated at 37 ° C. for 15 minutes. The reaction is stopped by adding the cell lysate attached to the cAMP measurement EIA kit (cAMP EIA (non-acetylation); GE Healthcare Bioscience Co., Ltd.), and intracellular cAMP according to the method described in the instruction manual. Concentration ([cAMP] i) was measured. The ability of the test substance to promote CGRP responsiveness was represented by [cAMP] i when stimulated with 0.5 nM CGRP alone as a reference (control), and the percentage was expressed as Improve Index. The CGRP responsiveness promoting ability of the test substance was tested for a significant difference from the control [cAMP] i by t-test.

(2) Results Table 2 shows the measurement results of CGRP responsiveness.

Example 2
The extract concentration dependency of CGRP responsiveness was examined.
Cells were prepared by the same procedure as in Example 1 except that the test substances shown in Table 1 were used in the amounts shown in Table 3, and CGRP responsiveness was measured. Table 3 shows the measurement results. The ability of the test substance to promote CGRP responsiveness was tested for significance against the control [cAMP] i by Dunnett's test.

Claims (4)

  Butcher Bloom, Yellowfin, Hemerocallis Fulva, Buttercup, Hibermata, Birch, Rice Protein, Rice Oil, Aloe Vera, Buttonpi, Laminaria, Star Fruit, Cornflower, Lemongrass, Kyonin, Lemon, Sweet Marjoram, Maronier, Royal Jelly, Rosehip and Their A cell CGRP responsive promoter comprising at least one of materials selected from the group consisting of extracts as an active ingredient.   The promoter according to claim 1, wherein the cells are vascular smooth muscle cells.   A method of promoting CGRP responsiveness of a cell, which has CGRP receptor and wants to promote CGRP responsiveness, includes butcher bloom, yellowfin, hemocalysfurba, spinach, hibermata, birch, rice protein, rice oil, Cultured in the presence of at least one material selected from the group consisting of aloe vera, buttonpi, laminaria, star fruit, cornflower, lemongrass, kyounin, lemon, sweet marjoram, maronier, royal jelly, rosehip and extracts thereof. A method comprising the step of:   4. The method of claim 3, wherein the cell is a vascular smooth muscle cell.