JP5730548B2 - CGRP response promoter - Google Patents

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 is also known to be involved in pregnancy and delivery due to its anti-inflammatory effect through prostaglandin synthesis, preconditioning for ischemia-reperfusion injury, and the expression of receptors in the myometrium, uterus, and placenta. ing.

  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 type hypersensitivity, pregnancy toxemia, premature labor and the like. Moreover, the treatment of migraine, hypersensitivity, etc. is mentioned as a use of a CGRP receptor antagonist. In fact, many CGRP receptor antagonist compounds have been developed as anti-migraine agents.

In addition, the relationship between CGRP and periodontal disease is also known. Non-Patent Document 6 shows that many sensory nerves expressing CGRP enter during inflammation, and CGRP exhibits anti-inflammatory through suppression of H ₂ O ₂ production and antigen presentation in macrophages and suppression of lymphocyte division. It has been reported. In addition, it has been reported that it is involved in dental pulp formation because it has the activity of promoting the growth of human dental pulp cells and is expected to form dentin through the promotion of BMP-2 expression (Non-patent Document 7). 8). Furthermore, since it has a bone resorption suppressing action (Non-Patent Document 9), periodontal disease is also mentioned as a therapeutic use of CGRP or a CGRP responsiveness promoter.

  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 Neuropeptides in Dental Pulp, 2008, 34 (7): 773-788 J Dent Res, 1995, 74: 1066-1071 J Endod, 1997, 23: 485-489 Calcif Tissue Int, 1987, 40: 149-154

  The present invention relates to the provision of a CGRP responsive promoter for cells containing carrot or an extract thereof as an active ingredient.

  When the present inventors examined the material which can accelerate | stimulate CGRP responsiveness of a cell, it discovered that CGRP responsiveness was accelerated | stimulated by the carrot extract.

That is, the present invention provides the following.
(1) A CGRP responsive promoter for cells containing carrot or an extract thereof as an active ingredient.
(2) The promoter according to (1), wherein the cell is a vascular smooth muscle cell.
(3) The promoter according to (1) or (2), wherein the extract is a water extract or an aqueous ethanol extract.
(4) A preventive and / or ameliorating agent for periodontal disease comprising carrot or an extract thereof as an active ingredient.
(5) A method for promoting CGRP responsiveness of a cell, comprising a step of culturing a cell having a CGRP receptor and desired to promote CGRP responsiveness in the presence of carrot or an extract thereof.
(6) The method according to (5), wherein the cell is a vascular smooth muscle cell.
(7) The method of (5) or (6), wherein the extract is a water extract or an aqueous ethanol extract.

  ADVANTAGE OF THE INVENTION According to this invention, the CGRP responsiveness promoter which can promote CGRP responsiveness of a cell is provided. The CGRP responsive 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 such as periodontal disease.

Promotion of CGRP responsiveness of HCASMC by carrot extract. N = 3. **: p <0.01 by Dunnett's significance test for [cAMP] i in wells containing only 0.5 nM CGRP. Promotion of CGRP responsiveness of HCASMC under various carrot extract addition conditions. N = 3, **: p <0.01 by Dunnett's significance test for [cAMP] i in wells (condition 1) 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 deterioration 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 the present specification, “carrot” refers to Panax ginseng belonging to the genus Arachnaceae , and “carrot extract” means an extract obtained from carrot. The extract can be any part of the carrot, such as whole grass, leaves, stems, buds, flowers, buds, wood parts, bark, lichens, roots, rhizomes, corms, corms, tubers, seeds, fruits, mycorrhiza or resin. Etc., or a combination thereof, but an extract from the root is preferred. The part may be subjected to the extraction step as it is, 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 carrot extract in the present invention can be obtained by extracting carrots 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 carrots (dry mass conversion). The extraction conditions are not particularly limited as long as sufficient extraction can be performed, but the extraction time is preferably 3 minutes to 30 days, more preferably 60 minutes to 14 days, and the extraction temperature is from 0 ° C to the boiling point of the solvent. Preferably, 5-70 degreeC is more preferable. 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 Examples described later, carrot or an extract thereof has an effect of significantly promoting the CGRP responsiveness of cells. Therefore, carrot or its extract is useful as a CGRP responsiveness promoter for cells. In addition, carrot or an extract thereof can exert an effect of improving skin circulation through the CGRP responsiveness promoting action, resulting in improvement of skin metabolism, promotion of blood circulation, promotion of wound healing, or hypertension, heart failure. Prevention of diseases such as liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor And / or useful for improvement.

  That is, carrot or an extract thereof can be used for promoting CGRP responsiveness of cells. Alternatively, carrots or extracts thereof can be used to improve skin circulation or skin metabolism, promote blood circulation, or promote wound healing. Or alternatively, carrots or extracts thereof may cause hypertension, heart failure, liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal It can be used for prevention and / or improvement of diseases such as illness, pregnancy toxemia and premature labor. Preferably, the disease is periodontal disease. The use can be in humans or non-human animals, or tissues, organs, cells derived therefrom, and can be therapeutic or non-therapeutic.

  Therefore, as one aspect, the present invention provides a CGRP responsive promoter for cells containing carrot or an extract thereof as an active ingredient. 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 responsive promoter of the present invention consists essentially of carrot or an extract thereof. Preferably, the disease is periodontal disease.

  As another aspect, the present invention provides a skin circulation improving agent containing carrot or an extract thereof as an active ingredient. As another aspect, the present invention provides a skin metabolism improving agent containing carrot or an extract thereof as an active ingredient. Preferably, the agent for improving skin metabolism is an agent for improving reduction in skin metabolism due to aging or the like. In one embodiment, these agents consist essentially of carrots or extracts thereof.

  As another aspect, the present invention provides a blood circulation promoter containing carrot or an extract thereof as an active ingredient. As another aspect, the present invention provides a wound healing promoter containing carrot or an extract thereof as an active ingredient. In yet another aspect, the present invention relates to hypertension, heart failure, ischemic reperfusion-induced liver injury, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage containing carrot or its extract as an active ingredient, Provided is a preventive and / or ameliorating agent for diseases such as pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, and premature labor. As a preferred embodiment, the present invention provides a preventive and / or ameliorating agent for periodontal disease comprising carrot or an extract thereof as an active ingredient. In one embodiment, these agents consist essentially of carrots or extracts thereof.

  Carrots or extracts thereof can enhance CGRP responsiveness of cells, improve skin circulation or skin metabolism, promote blood circulation, promote wound healing, or hypertension, heart failure, ischemia Prevention and / or prevention of diseases such as liver damage due to reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor It can be used for the production of compositions for improvement, medicines, quasi drugs, cosmetics, foods and drinks and the like. 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 carrot or its extract or the CGRP responsive promoter of the present invention is blended in the composition, medicine, quasi-drug, cosmetics, food and drink, etc., and improves skin circulation for promoting CGRP responsiveness of cells. Or for improving skin metabolism, for promoting blood circulation, for promoting wound healing, or for hypertension, heart failure, liver damage due to ischemic reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, lung It may be an active ingredient for preventing and / or improving diseases such as essential hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, and premature labor.

The said pharmaceutical or quasi-drug contains the carrot or its extract, or the CGRP responsive promoter of this 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.
Preferably, the medicine or quasi-drug is a dentifrice, mouthwash, mouthspray, mouthwash, or oral ointment containing carrot or an extract thereof or the CGRP response promoter of the present invention as an active ingredient. Oral compositions such as creams and coating agents can be used.

  The medicament or quasi-drug may contain carrot or an extract thereof or the CGRP responsive promoter of the present invention alone or in combination with a pharmaceutically acceptable 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 the other active ingredient and pharmacological component, as long as the CGRP responsive acceleration | stimulation effect | action of a carrot or its extract is not lost.

  The pharmaceutical or quasi-drug is prepared from a carrot or an extract thereof or the CGRP responsive promoter of the present invention, or in combination with the carrier and / or other active ingredients and pharmacological ingredients as necessary. Can be manufactured. The content of the carrot or its extract in the medicine or quasi-drug is preferably 0.00001 to 100% by mass, more preferably 0.00002 to 100% by mass, as a dry weight. More preferably, the content is 0.0005 to 100% by mass, even more preferably 0.0001 to 100% by mass, and still more preferably 0.001 to 100% by mass.

  The said food-drinks contain the carrot or its extract, or the CGRP responsiveness promoter of this invention as an active ingredient. The food and drink is based on the concept of promoting CGRP responsiveness and preventing and / or improving periodontal disease, and if necessary, the food and drink, functional food, food for the sick, and food for specified health use. be able to.

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.
As a preferred form, the food and drink of the present invention may be gum, candy, tablet, beverage, pet gum, etc. with the concept of periodontal disease prevention and / or improvement.

In order to prepare the above-mentioned various forms of food and drink, the carrot or its extract or the CGRP response promoter 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 carrot or the extract thereof in the food or drink is preferably 0.00001 to 100% by mass, more preferably 0.00002 to 75% by mass, as dry weight, and 0.00005 to The content is more preferably 50% by mass, even more preferably 0.0001 to 30% by mass, and still more preferably 0.0005 to 20% by mass.

  The cosmetic contains carrot or an extract thereof or the CGRP responsiveness promoter of the present invention as an active ingredient. The cosmetic may contain carrot or an extract thereof or the CGRP responsiveness promoter of the present invention alone, or may be contained in combination with a carrier acceptable as a cosmetic. 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 carrots or extracts thereof is not lost, the cosmetics are other active ingredients and cosmetic ingredients such as moisturizers, whitening agents, UV protection agents, cell activators, detergents , Keratolytic agents, makeup ingredients (for example, makeup bases, foundations, funny, powders, teak, lipsticks, eye makeup, eyebrow, mascara, etc.) and the like. 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 carrots or extracts thereof or the CGRP responsive promoter of the present invention or, if necessary, in combination with the carrier and / or other active ingredients and cosmetic ingredients by a conventional method. it can. The content of the carrot or its extract in the cosmetic is preferably 0.00001 to 100% by mass, more preferably 0.00002 to 50% by mass, and more preferably 0.005 to 30% as a dry weight. It is more preferable to set it as the mass%, It is still more preferable to set it as 0.0001-20 mass%, It is still more preferable to set it as 0.001-10 weight%.

  The present invention also provides a method for promoting CGRP responsiveness of cells. The method includes the step of adding carrot or an extract thereof to a cell having a CGRP receptor and in which CGRP responsiveness is desired to be promoted. When the cell is a cell culture, the concentration of the added carrot or its extract is 0.00001 to 2% (w / v), preferably 0.0001 to 1% (w / v) as a dry weight. And more preferably 0.0003 to 0.8% (w / v). The timing of adding the carrot or an extract thereof may be before CGRP stimulation and / or at the time of CGRP stimulation.

The carrot or its extract 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, heart failure. Prevention of diseases such as liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor And / or for improvement, can be administered in an effective amount to a subject in need thereof.
In one embodiment, the administration is non-therapeutically intended to improve skin circulation, improve skin metabolism, or promote 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. Preferably, an animal suffering from periodontal disease, an animal suspected thereof, or an animal having a high risk thereof is mentioned as an administration target. 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 6000 mg / day, more preferably 100 μg to 3000 mg / day, more preferably 1000 μg to 2000 mg / day, in terms of carrot bulk drug. Preferably, 10,000 μg to 1000 mg / day is more preferable.

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

Production Example 1
1000 g of 50% ethanol aqueous solution was added to 100 g of ginseng root Panax ginseng (made in China), soaked and extracted at room temperature for 7 days, and then filtered to obtain 840 mL of carrot extract (evaporation residue 2.8 w / v%).

Example 1
The effect of promoting CGRP responsiveness of cells by carrot extract was examined.
(1) Method 1. 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. Cell Preparation Normal human coronary artery smooth muscle cells were seeded in a 48-well plate at a density of 4000 cells / cm ² using a growth medium, 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 two days, the cells were cultured in a differentiation medium containing carrot extract (Ichimaru Falcos Co., Ltd., Singhongginisin LV, extract content 2.8 w / v%) shown in Table 1 and 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.) at a predetermined concentration. The cells were washed twice with a basal medium containing carrot extract (Humedia-SB2; Kurabo Industries), 200 μl of the same medium was further added, and cultured at 37 ° C. for 60 minutes to allow the inhibitor to permeate the cells. The medium was removed, a basal medium containing carrot extract, 0.5 nM CGRP, and phosphodiesterase inhibitor at the concentrations shown in Table 1 was added, and cultured 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 to promote CGRP responsiveness of carrot extract 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.

(2) Result The measurement result of CGRP responsiveness is shown in FIG. Improve Index 100% corresponds to [cAMP] i of 893.5 fmol / well. The Improve Index increased according to the carrot extract concentration.

Example 2
The CGRP responsiveness of the cells was examined by variously changing the carrot extract addition conditions.
(1) Method HCASMC (Kurabo, cell lot # 625723) was prepared under the same conditions as in Example 1. The last two days of culture in the differentiation medium were cultured under the conditions shown in Table 1, and then the CGRP responsiveness of the cells was measured in the same manner as in Example 1.

(2) Result The measurement result of CGRP responsiveness is shown in FIG. Improve Index 100% corresponds to [cAMP] i of 893.5 fmol / well. Improve Index increased by adding carrot extract during the last 2 days of culture in differentiation medium to CGRP stimulation (conditions 2-4). On the other hand, since the Improve Index was significantly low in the absence of CGRP (condition 5), the action of carrot extract is not a substitute for CGRP (for example, a CGRP receptor agonist) but CGRP and its receptor. It has been shown to promote a series of responses elicited by binding.

Claims (6)

A CGRP responsive promoter for cells containing carrots or extracts thereof as an active ingredient (except when used as a food or drink, a preventive and / or ameliorating agent for periodontal disease, or a blood circulation promoter) .   The promoter according to claim 1, wherein the cells are vascular smooth muscle cells.   The promoter according to claim 1 or 2, wherein the extract is a water extract or an aqueous ethanol extract.   A method for promoting CGRP responsiveness of a cell, comprising culturing a cell having a CGRP receptor and desired to promote CGRP responsiveness in the presence of carrot or an extract thereof. The method of claim 4 , wherein the cell is a vascular smooth muscle cell. The method according to claim 4 or 5 , wherein the extract is a water extract or an aqueous ethanol extract.

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