JP2012116766A - Cgrp responsiveness suppressor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a material for suppressing CGRP responsiveness of cells.SOLUTION: There is provided the CGRP responsiveness suppressor of cells containing at least one substance selected from the group consisting of great burnet, meadow saxifrage, evening primrose, Himalayan raspberry and their extracts as an active component.

Description

  The present invention relates to a CGRP (calcitonin gene-related peptide) responsive inhibitor.

  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 most well-known physiological action of CGRP is a vasodilating action and an accompanying increase in blood flow, and is considered to be involved in skin blood flow regulation. CGRP administration increases skin blood flow in rats (Non-patent document 1), CGRP knockout mice have increased blood pressure (Non-patent document 2), and rats administered CGRP neutralizing antibody have decreased skin blood flow. (Non-patent Document 3) In patients with Raynaud's disease that is thought to be caused by excessive contraction of peripheral blood vessels, CGRP-containing nerves in fingers are lacking (Non-patent Document 4). It has been known that the skin blood flow of the arm is increased by intravenous injection (Non-patent Document 5).

  CGRP is also involved in cerebrovascular disorders such as migraine and cluster headache. High levels of CGRP in the jugular vein of patients with migraine attacks (Non-patent document 6), and migraine patients with elevated CGRP levels in saliva during the period between attacks (Non-patent document 7) CGRP itself triggers migraine (Non-patent document 8), CGRP antagonist BIBN4096BS is effective in treating acute migraine attacks (Non-patent document 9), CGRP injecting headache induced by CGRP infusion It has been reported that it can be prevented by administration (Non-patent Document 10).

Various other actions have been reported for the physiological action of CGRP. For example, regulation of substance P in inflammation, increased vascular permeability, regulation of nicotinic receptors at the neuromuscular junction, inhibition of gluconeogenesis and promotion of glycolysis, promotion of pancreatic enzyme secretion, inhibition of gastric acid secretion, promotion of heart rate, nerve Various activities such as regulation of activity, regulation of calcium metabolism, promotion of bone formation, insulin secretion, increase in body temperature, decrease in food intake. 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 changes in skin circulation and preventing and / or treating the above diseases and symptoms. So far, CGRP receptor antagonists (Non-Patent Documents 9 and 10), anti-CGRP antibodies, and Iris genus water extract (Patent Document 1) are known as substances that can suppress CGRP responsiveness. However, it is desired to develop further substances that can promote or suppress CGRP responsiveness and are useful for improving various diseases associated with CGRP.

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 Ann Neurol, 1990, 28: 183-187 Headache, 2006, 46: 24-33 Cephalalgia, 2002, 22: 54-61 New Engl J Med, 2004, 350: 1104-1110 Clin Pharmacol Ther, 2005, 77: 202-213

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

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

That is, the present invention provides the following.
(1) A CGRP responsive inhibitor for cells, comprising as an active ingredient at least one selected from the group consisting of bitumen, yukinoshita, evening primrose, Himalayan raspberry, and extracts thereof.
(2) The inhibitor according to (1), wherein the cell is a vascular smooth muscle cell.
(3) A prophylactic and / or ameliorating agent for migraine, comprising at least one selected from the group consisting of walnut mushrooms, saxifrage, evening primrose, Himalayan raspberry, and extracts thereof.
(4) A prophylactic and / or ameliorating agent for hypersensitivity comprising at least one selected from the group consisting of walnut mushrooms, saxifrage, evening primrose, Himalayan raspberries and extracts thereof as an active ingredient.
(5) A method for suppressing CGRP responsiveness of a cell, wherein a cell having a CGRP receptor and which is desired to suppress CGRP responsiveness is selected from the group consisting of burdock, saxifrage, evening primrose, Himalayan raspberry and extracts thereof. Culturing in the presence of at least one of the selected materials.
(6) The method according to (5), wherein the cell is a vascular smooth muscle cell.

  ADVANTAGE OF THE INVENTION According to this invention, the CGRP responsiveness inhibitor which can suppress the CGRP responsiveness of a cell is provided. The CGRP responsive inhibitor is useful for the prevention and / or treatment of various diseases and symptoms related to CGRP.

Inhibition of CGRP responsiveness of HCASMC by bitumen extract. N = 3. **: p <0.01 by Dunnett's significance test for [cAMP] i in wells containing only 0.5 nM CGRP. Inhibition of CGRP responsiveness of HCASMC by Yukinoshita extract. N = 3. **: p <0.01 by Dunnett's significance test for [cAMP] i in wells containing only 0.5 nM CGRP. Inhibition of CGRP responsiveness of HCASMC by evening primrose extract. N = 3. **: p <0.01 by Dunnett's significance test for [cAMP] i in wells containing only 0.5 nM CGRP. Inhibition of CGRP responsiveness of HCASMC by Himalayan raspberry extract. N = 3. **: p <0.01 by Dunnett's significance test for [cAMP] i in wells containing only 0.5 nM 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, “inhibition of CGRP responsiveness of cells” refers to suppression of 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, vascular smooth muscle cells open K ⁺ channels through protein kinase A activation. Alternatively, in vascular endothelial cells, eNOS is activated through protein kinase A activation, and production of NO 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 inhibitor according to the present invention can suppress the activation of these series of processes in cells induced by CGRP. For example, the action of the CGRP responsive inhibitor of the present invention includes inhibition of binding between CGRP and its receptor, suppression of G protein activation, suppression of adenylate cyclase activation, suppression of increase in intracellular cAMP, Examples include inhibition of protein kinase A activation, inhibition of NO production or cGMP activation, and inhibition of K ⁺ channel opening. Preferably, the “cell CGRP responsiveness” can be determined using the degree of suppression of the increase in intracellular cAMP caused by CGRP as an index. Therefore, the CGRP responsive inhibitor of the present invention preferably suppresses the increase in intracellular cAMP caused by CGRP.

In the present invention, the “cell” that suppresses 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 the present specification, “Wale Moko” refers to Sanguisorba officinalis of the Rosaceae family, “Yukinoshita” refers to Saxifraga stolonifera of the Family of Sphagnumaceae , “Erica evening primrose” refers to Oenothera biennis of the red clover family, “Himalayan raspberry” "Refers to Rubus ellipticus of the Rosaceae family.

  Unless specifically limited, the plant extract is any part of the plant, for example, whole grass, leaves, stems, buds, flowers, buds, wood parts, bark, lichens, roots, rhizomes, corms, corms, tubers , Seeds, fruits, mycorrhiza, resin, etc., or combinations thereof. Preferred sites for obtaining the extract are the roots and rhizomes for walnut mushrooms, whole grass for saxifrage, seeds for evening primrose, and roots for Himalayan raspberry. 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 plants (dry mass conversion). The extraction conditions are not particularly limited as long as the extraction is performed at a temperature between 0 ° C. and the boiling point of the solvent so that sufficient extraction can be performed. 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.
Furthermore, from the viewpoint of improving the pharmacological activity of the extract, separation / purification techniques such as chromatography and liquid-liquid partition may be used to remove inactive impurities from the extract.

  As shown in Examples described later, a material selected from the group consisting of walnut mushrooms, snowy shore, evening primrose, Himalayan raspberry, and extracts thereof has an effect of significantly suppressing the CGRP responsiveness of cells. Therefore, these materials are useful as a CGRP response inhibitor of cells. In addition, these materials can exert an effect of preventing and / or improving diseases such as migraine and hypersensitivity through the CGRP responsiveness suppressing action.

  That is, the above material can be used for suppressing CGRP responsiveness of cells, or for preventing and / or improving diseases such as migraine and hypersensitivity. 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.

  Therefore, as one aspect, the present invention provides a CGRP responsive inhibitor for cells containing, as an active ingredient, at least one selected from the group consisting of walnut mushrooms, saxifrage, evening primrose, Himalayan raspberry, and extracts thereof. . The CGRP responsive inhibitor can be used for suppressing CGRP responsiveness of cells, or for preventing and / or improving diseases such as migraine and hypersensitivity. As one embodiment, the CGRP responsiveness inhibitor of the present invention consists essentially of at least one of the above materials.

  As another aspect, the present invention relates to a disease such as migraine, hypersensitivity, etc., containing as an active ingredient at least one material selected from the group consisting of bituminous flowers, saxifrage, evening primrose, Himalayan raspberry and extracts thereof. An agent for preventing and / or improving the above is provided. In one embodiment, the agent consists essentially of at least one of the materials.

  A material selected from the group consisting of bituminous, cypress, evening primrose, Himalayan raspberry, and extracts thereof is for suppressing CGRP responsiveness of cells or for preventing and / or improving diseases such as migraine and hypersensitivity. It can be used for the production of a composition, a medicine, a quasi-drug, a cosmetic, a food and drink. 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 responsive inhibitor of the present invention is blended in the composition, medicine, quasi-drug, cosmetics, food and drink, etc., for suppressing CGRP responsiveness of cells, or migraine It may be an active ingredient for preventing and / or improving diseases such as hypersensitivity.

  The medicine or quasi-drug contains at least one of the above-mentioned materials or the CGRP responsiveness inhibitor 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 inhibitor 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 a pharmacological component, as long as the CGRP responsiveness suppression 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 inhibitor of the present invention, or in combination with the above carrier and / or other active ingredients and pharmacological ingredients as necessary. Can be manufactured. The content of the material in the medicine or quasi-drug is preferably 0.000001 to 100% by mass, more preferably 0.000002 to 100% by mass, as dry weight, and 0.000005 to More preferably, the content is 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-drinks contain the raw material mentioned above or the CGRP responsive inhibitor of this invention as an active ingredient. The food and drink is based on the concept of CGRP responsive suppression, migraine, prevention and / or improvement of diseases such as hypersensitivity, etc. It can be a food or a food for specified health use.

  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-mentioned materials or the CGRP responsiveness inhibitor 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 above-mentioned materials in the food and 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 inhibitor of the present invention as an active ingredient. The cosmetic may contain the above-mentioned materials or the CGRP responsiveness inhibitor 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 do it. 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 suppressing action of the above materials is not lost, the cosmetics are effective 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 is produced from the above-mentioned materials or the CGRP responsiveness inhibitor of the present invention or, if necessary, by combining the carrier and / or other active ingredients and cosmetic ingredients according to a conventional method. Can do. The content of the material in the cosmetic is preferably 0.000001 to 100% by mass, more preferably 0.000002 to 50% by mass, and 0.000005 to 30% by mass as a dry weight. More preferably, it is more preferably 0.00001 to 20% by mass, and still more preferably 0.0001 to 10% by weight.

  The present invention also provides a method for suppressing CGRP responsiveness of cells. In the method, at least one of materials selected from the group consisting of walnut mushrooms, saxifrage, evening primrose, Himalayan raspberry, and extracts thereof is added to cells that have a CGRP receptor and want to suppress CGRP responsiveness. Including a process. When the cell is a cell culture, the concentration of the material added is 0.000001 to 2% (w / v), preferably 0.00001 to 1% (w / v) as a dry weight. More preferably, it is 0.00003-0.8% (w / v). The timing of adding the above-mentioned materials may be before CGRP stimulation and / or during CGRP stimulation.

  The above-mentioned materials or the CGRP responsive inhibitor of the present invention is used for the suppression of CGRP responsiveness, the improvement of skin circulation or the improvement of skin metabolism, or the prevention and / or improvement of diseases such as migraine and hypersensitivity. Thus, they can be administered in an effective amount to a subject in need thereof.

Examples of the administration target include animals suffering from diseases such as migraine and hypersensitivity, animals suspected to be high risk, and animals having 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 orally administered to humans, the dose is preferably 10 μg to 50000 mg / day per adult as the dry weight of walnut mushroom, snowy night, evening primrose, Himalayan raspberry or an extract thereof, and 100 μg to 30000 mg / day. The day is more preferable, 1000 μg to 20000 mg / day is more preferable, and 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 Cell CGRP Responsiveness Inhibitory Effect by Various Substances (1) Method Cell culture Normal human coronary artery smooth muscle cells (HCASMC; Kurabo Industries, 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) 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 suppress 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 an Inhibition Index. The ability of the test substance to suppress CGRP responsiveness was tested for a significant difference with respect to the control [cAMP] i, and a p value of less than 0.1 was considered to be significantly effective.

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

Example 2 Inhibition of CGRP responsiveness of cells by bituminous extract Cellulose extract of Table 1 was used as a test substance at different amounts, and the cells were cultured by the same method as in Example 1, and the CGRP responsiveness of the cells was measured. . The results are shown in FIG. Inhibition Index 100% corresponds to [cAMP] i of 1306.5 fmol / well.

Example 3 Inhibition of CGRP responsiveness of cells by Yukinosita extract Using different amounts of Yukinosita extract shown in Table 1 as a test substance, cells were cultured by the same method as in Example 1, and the CGRP responsiveness of the cells was measured. . The results are shown in FIG. Inhibition Index 100% corresponds to [cAMP] i of 1256.0 fmol / well.

Example 4 Inhibition of CGRP responsiveness of cells by evening primrose extract Using different amount of evening primrose extract shown in Table 1 as a test substance, cells were cultured by the same method as in Example 1, and the CGRP responsiveness of the cells was measured. . The results are shown in FIG. Inhibition Index 100% corresponds to [cAMP] i of 1306.5 fmol / well.

Example 5 Inhibition of CGRP responsiveness of cells by Himalayan raspberry extract Using different amounts of Himalayan raspberry extract described in Table 1 as a test substance, cells were cultured by the same method as in Example 1, and the CGRP responsiveness of the cells was measured. It was measured. The results are shown in FIG. Inhibition Index 100% corresponds to [cAMP] i of 1306.5 fmol / well.

Claims (6)

  A CGRP responsive inhibitor of a cell comprising as an active ingredient at least one of materials selected from the group consisting of bituminous plant, snowy mushroom, evening primrose, Himalayan raspberry and extracts thereof.   The inhibitor according to claim 1, wherein the cell is a vascular smooth muscle cell.   A prophylactic and / or ameliorating agent for migraine comprising, as an active ingredient, at least one selected from the group consisting of bitumou, yukinoshita, evening primrose, Himalayan raspberry and extracts thereof.   A prophylactic and / or ameliorating agent for hypersensitivity comprising at least one selected from the group consisting of walmocow, saxifrage, evening primrose, Himalayan raspberry and extracts thereof as an active ingredient.   A method for suppressing CGRP responsiveness of a cell, wherein a cell having a CGRP receptor and desired to suppress CGRP responsiveness is selected from the group consisting of walnut mushrooms, saxifrage, evening primrose, Himalayan raspberry and extracts thereof Culturing in the presence of at least one of the materials.   6. The method of claim 5, wherein the cell is a vascular smooth muscle cell.

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