JP6163359B2 - 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. This vascular network creates skin circulation. 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 axonal reflex. 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 and peripheral nerves and exerts its action through specific receptors. It is well known that CGRP receptors are expressed in vascular endothelial cells and vascular smooth muscle cells. In any cell, the blood vessel dilates and blood flow increases when CGRP binds to its receptor. In vascular endothelial cells, when CGRP binds to a cellular CGRP receptor, an increase in intracellular cAMP occurs through activation of adenylate cyclase via the G protein. The increase in cAMP activates protein kinase A, which in turn activates eNOS, thus promoting NO production. The produced NO acts on vascular smooth muscle cells, and K ⁺ channels are opened through intracellular cGMP activation via NO. This relaxes the vascular smooth muscle and causes vascular dilation, resulting in increased blood flow. When CGRP directly binds to vascular smooth muscle cells, protein kinase A is activated by the activation of adenylate cyclase via G protein, and the K ⁺ channel is opened. As a result, the vascular smooth muscle is also relaxed, the blood vessels are dilated, and the blood flow is increased.

  The relationship between CGRP and blood flow is well known. For example, the administration of CGRP in rats increases skin blood flow (Non-Patent Document 1), the blood pressure of CGRP knockout mice increases (Non-Patent Document 2), and rats administered with a CGRP neutralizing antibody exhibit skin blood flow. In patients with Raynaud's disease that is thought to be due to excessive contraction of peripheral blood vessels (Non-patent document 4), patients with Raynaud's disease It is known that the skin blood flow of the arm is increased by intravenously injecting CGRP (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.

  From the above physiological effects, CGRP or CCRP responsive promoters can be used for blood circulation promotion, blood flow promotion, wound healing promotion, and hypertension, heart failure, liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, Treatment of sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed type hypersensitivity, pregnancy toxemia, premature labor and the like are mentioned. 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. Moreover, since it has the activity of promoting the growth of human dental pulp cells and the formation of dentine through the promotion of the expression of BMP-2, it is reported that it is involved in the formation of dental pulp (Non-patent Document 7). 8). Furthermore, periodontal disease is also mentioned as a therapeutic use of CGRP or a CGRP response promoter because of its bone resorption inhibitory effect (Non-Patent Document 9).

  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.

  Panax ginseng is a medicinal plant known for a long time. Ginsenoside is known as a medicinal component, and 50 or more types of ginsenoside have been isolated from ginseng so far. In addition to the well-known tonic effect of ginseng or ginsenoside, in recent years antitumor action, anti-arteriosclerosis / antihypertensive action, anti-stress action, immunoregulatory action, anti-inflammatory / anti-allergic action, anti-diabetes mellitus Various effects such as action, action on central nervous system, improvement of memory and learning, neuroprotection, neurotransmitter release or uptake action have been found (Non-patent Documents 10 and 11). In addition, other ginsenoside activities include voltage-dependent Ca channel inhibition, calcium-dependent K channel activation, ATP-sensitive K channel activation, G protein-coupled inward rectifier K channel activation, voltage-dependent Na channel inhibition, Effects on ion channels such as inhibition of NMDA agonistic ion channels, inhibition of nicotinic acetylcholinergic ion channels, and inhibition of serotonergic ion channels have been reported (Non-patent Document 12).

  However, there was no report that ginsenoside is involved in the control of CGRP responsiveness.

Japanese 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 Journal of Endodontics, 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 Advances in Food and Nutrition Research, 2009, 55: 1-99 Phytochemical Analysis, 2008, 19: 2-16 CNS Drug Reviews, 2007, 13 (4): 381-404

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

  The present inventors have examined materials that can promote CGRP responsiveness of cells, and have found that a ginsenoside compound having a specific structure has a CGRP responsiveness promoting action.

That is, the present invention provides the following.
(1) The following formula (I):
(In the formula (I),
R ₁ is hydrogen or a sugar residue consisting of 1 to 3 monosaccharide units;
R ₂ is a group represented by the following formula (i) or formula (ii);
In formula (i), R ₃ is hydrogen or a sugar residue consisting of 1 to 3 monosaccharide units; in formula (ii), either a or b is a bond;
In each of R ₁ and R _3, the sugar residue is linear or branched; the 1 to 3 monosaccharide units are each independently composed of glucose, xylose, arabinose, rhamnose and glucuronic acid. Selected from the group and each independently substituted or unsubstituted)
Or the following formula (II):
A CGRP responsive promoter for cells, which comprises a compound represented by formula (I) or an optical isomer thereof as an active ingredient.
(2) A preventive and / or ameliorating agent for periodontal disease comprising the compound represented by the formula (I) or (II) or an optical isomer thereof as an active ingredient.
(3) A blood flow and / or blood circulation promoter comprising the compound represented by the formula (I) or (II) or an optical isomer thereof as an active ingredient.
(4) A method for promoting CGRP responsiveness of a cell, the method comprising culturing a cell having a CGRP receptor and desired to promote CGRP responsiveness in the presence of the CGRP responsive promoter described in (1) Including a method.

  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 used to improve changes in skin circulation due to aging, etc., to promote blood flow and / or blood circulation, and to prevent or treat various diseases and symptoms related to CGRP such as periodontal disease. Useful.

Promotion of CGRP responsiveness of HCASMC by ginsenoside compound. *: P <0.05, **: p <0.01 (vs. control, Dunnett test, N = 3). Promotion of CGRP responsiveness of HCASMC by ginsenoside compound. *: P <0.05, **: p <0.01 (vs. control, Dunnett test, N = 3). Promotion of CGRP responsiveness of HCASMC by ginsenoside compounds added under different conditions. *: P <0.05, **: p <0.01 (vs. control, Dunnett test, N = 3).

  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. This relaxes the vascular smooth muscle and causes vascular dilation, resulting in increased blood flow. 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. As a result, vascular smooth muscle relaxes, vascular dilation occurs, and blood flow increases.
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.

  The cell CGRP responsive promoter, the preventive and / or ameliorant of periodontal disease, the blood flow and / or the blood circulation promoter according to the present invention include a compound represented by the following formula (I) or an optical isomer thereof: The active ingredient.

In formula (I), R ₁ is hydrogen or a sugar residue consisting of 1 to 3 monosaccharide units, preferably hydrogen or a sugar residue consisting of 1 to 2 substituted or unsubstituted monosaccharide units. It is a group.
In formula (I), R ₂ is a group represented by the following formula (i) or formula (ii), or an optical isomer of formula (i).

In the formula (i), R ₃ is hydrogen or a sugar residue consisting of 1 to 3 monosaccharide units, preferably hydrogen or a sugar residue consisting of 1 to 2 substituted or unsubstituted monosaccharide units. It is. In formula (ii), one of a and b is a bond.

In each of R ₁ and R ₃ , the sugar residue may be linear or branched. In each of R ₁ and R _3, the monosaccharide unit constituting the sugar residue is independently selected from the group consisting of glucose, xylose, arabinose, rhamnose, and glucuronic acid. Preferably, the monosaccharide unit of R ₁ is glucose and the monosaccharide unit of R ₃ is glucose or arabinose.

  The monosaccharide units can also be independently substituted or unsubstituted. Preferably each monosaccharide unit is unsubstituted, more preferably all monosaccharide units are unsubstituted. When the monosaccharide unit is substituted, the monosaccharide unit located at the end of the sugar residue (that is, when the sugar residue consists of one monosaccharide unit, the monosaccharide unit In the case of consisting of a monosaccharide unit, it is preferable that only the monosaccharide unit located at the end not bonded to the oxygen atom in the above formula (I) or formula (i) is substituted.

Examples of the substituent when the monosaccharide unit is substituted include an acetyl group (Ac), a malonyl group, a butenoyl group, an octenoyl group, and an oleanoyl group with respect to the monosaccharide unit in R ₁ . Further, with respect to the monosaccharide units of R _3, acetyl groups. In both R ₁ and R ₃ , these substituents are preferably a monosaccharide unit located at the end of the sugar residue (that is, the monosaccharide unit when the sugar residue is composed of one monosaccharide unit). Or when the sugar residue is composed of a plurality of monosaccharide units, it is substituted at the 6-position of the above-mentioned formula (I) or the monosaccharide unit located at the end not bonded to the oxygen atom in formula (i)) The

Preferred examples of R ₁ include hydrogen, β-D-glucopyranosyl, 2-O- (β-D-glucopyranosyl) -β-D-glucopyranosyl, 2-O- (6-O-acetyl-β-D-glucopyranosyl). ) -Β-D-glucopyranosyl and the like.

Examples of preferable sugar residues for R ₃ include Glc-Glc-, Ara-Glc-, Xyl-Glc-, and the like. Preferred examples of R ₃ include hydrogen, β-D-glucopyranosyl, 6-O- (β-D-glucopyranosyl) -β-D-glucopyranosyl, 6-O- (α-L-arabinofuranosyl) -β. -D-glucopyranosyl or 6-O- (α-D-arabinopyranosyl) -β-D-glucopyranosyl and the like can be mentioned.

  The cell CGRP responsive promoter, the preventive and / or ameliorating agent for periodontal disease, the blood flow and / or blood circulation promoter according to the present invention is a compound represented by the following formula (II), or an optical isomer thereof: Is the active ingredient.

  In the present invention, a compound represented by the formula (I) or an optical isomer thereof (hereinafter collectively referred to as a compound of the formula (I)), and a compound represented by the formula (II) or an optical isomer thereof ( In the present specification, preferred examples of the compound (hereinafter collectively referred to as the compound of formula (II)) include the compounds exemplified in the following table.

  The compound of the formula (I) or the formula (II) can be isolated or purified from Panax ginseng (Panax ginseng belonging to the genus Tochibanin). For example, an extract extracted from a root of Panax ginseng with a solvent such as an aqueous ethanol solution is separated by column chromatography using an aromatic column such as Diaion HP20 or an HPLC ODS column, thereby obtaining the formula (I) Alternatively, the compound of formula (II) can be isolated. Alternatively, a commercially available ginsenoside compound may be used. For example, the compounds described in Table 1 above can be purchased from Carbosynth, LKT Laboratories, or the like.

  As shown in Examples described later, the compound of formula (I) or formula (II) has an effect of significantly promoting CGRP responsiveness of cells. Therefore, the compound of the formula (I) or the formula (II) is useful as a CGRP response promoter for cells. Further, the compound of formula (I) or formula (II) exerts effects such as improvement of skin circulation, promotion of blood flow and / or blood circulation, and promotion of dental pulp formation through the CGRP responsiveness promoting action. As a result, improvement of skin metabolism, promotion of blood flow and / or blood circulation, promotion of wound healing, or hypertension, heart failure, liver damage due to ischemic reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, It is useful for the prevention and / or improvement of diseases such as subarachnoid hemorrhage, pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, premature labor.

  That is, the compound of formula (I) or formula (II) can be used for promoting CGRP responsiveness of cells. Alternatively, the compound of formula (I) or formula (II) can be used to improve skin circulation or skin metabolism, to promote blood flow and / or blood circulation, to promote wound healing. Alternatively, the compound of formula (I) or formula (II) may be used for hypertension, heart failure, liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed It can be used for prevention and / or amelioration of diseases such as type hypersensitivity, periodontal disease, pregnancy toxemia, and premature labor. Preferably, the disease is periodontal disease. The use can be in human or non-human animals, or in vitro in tissues, organs, cells, etc. derived therefrom. In addition, the use may be a therapeutic use, or a non-therapeutic use intended to improve skin circulation for the purpose of promoting health or cosmetics, improving skin metabolism, or promoting blood flow and / or blood circulation. Also good. When the compound of the formula (I) or the formula (II) is used in the present invention, any one of the compounds represented by the formula (I) or the formula (II) described above or optical isomers thereof is used alone. Or any two or more of them may be used in combination.

  Therefore, as one aspect, the present invention provides a CGRP responsive promoter for cells containing the compound of formula (I) or formula (II) as an active ingredient. The CGRP responsive promoter can be used to promote CGRP responsiveness of cells, and based on the CGRP responsive promoting action, to improve skin circulation or skin metabolism, blood flow and / or To promote blood circulation, promote wound healing, or hypertension, heart failure, liver damage due to ischemia reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity It can be used for prevention and / or amelioration of diseases such as illness, periodontal disease, pregnancy toxemia, and premature labor. In one embodiment, the CGRP responsive promoter of the present invention consists essentially of a compound of formula (I) or formula (II). Preferably, the disease is periodontal disease.

  As another aspect, the present invention provides a skin circulation improving agent containing a compound of formula (I) or formula (II) as an active ingredient. In another aspect, the present invention provides a skin metabolism improving agent containing a compound of formula (I) or formula (II) 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 a compound of formula (I) or formula (II).

  In another aspect, the present invention provides a blood flow and / or blood flow promoter containing a compound of formula (I) or formula (II) as an active ingredient. In another aspect, the present invention provides a wound healing promoter containing a compound of formula (I) or formula (II) as an active ingredient. In still another aspect, the present invention relates to hypertension, heart failure, ischemic reperfusion-induced liver damage, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, containing the compound of formula (I) or formula (II) as an active ingredient. Provided is a preventive and / or ameliorating agent for diseases such as arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, premature labor. As a preferred embodiment, the present invention provides a preventive and / or ameliorating agent for periodontal disease comprising a compound of formula (I) or formula (II) as an active ingredient. In one embodiment, these agents consist essentially of a compound of formula (I) or formula (II).

  In another aspect, the present invention provides the use of a compound of formula (I) or formula (II) for the manufacture of a cellular CGRP responsive promoter. As yet another aspect, the present invention relates to a skin circulation improving agent, a blood flow and / or blood circulation promoter, a wound healing promoter, or hypertension, heart failure, liver damage caused by ischemia reperfusion, renal failure, gastrointestinal ulcer, Formula (I) or Formula for the manufacture of an agent for preventing and / or improving diseases such as sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor Use of a compound of (II) is provided. Preferably, the agent for improving skin metabolism is an agent for reducing skin metabolism due to aging or the like. Also preferably, the disease is periodontal disease. In one embodiment, the skin circulation improving action, the blood flow and / or blood circulation promoting action, the wound healing promoting action, or the disease preventing and / or improving action of the agent is brought about by a cell CGRP responsive promoting action. It is.

  The compound of formula (I) or formula (II) is a wound healing agent for promoting cell CGRP responsiveness, for improving skin circulation or for improving skin metabolism, for promoting blood flow and / or blood circulation. Hypertension, 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 poisoning It can be used for the production of a composition, medicine, quasi-drug, cosmetic, food and drink for the prevention and / or improvement of diseases such as illness and premature labor. 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 compound of formula (I) or formula (II) is blended in the composition, medicine, quasi-drug, cosmetics, food and drink, etc., and improves skin circulation or skin metabolism to promote CGRP responsiveness of cells. To improve, promote blood flow and / or blood circulation, promote wound healing, or hypertension, heart failure, liver damage due to ischemic reperfusion, renal failure, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid It may be an active ingredient for preventing and / or improving diseases such as bleeding, pulmonary hypertension, delayed type hypersensitivity, periodontal disease, pregnancy toxemia, and premature labor.

The said pharmaceutical or quasi-drug contains the compound of Formula (I) or Formula (II) 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 elixirs, syrups and suspensions. Examples of the dosage form for oral administration include injection, infusion, transdermal, transmucosal, nasal, enteral, inhalation, suppository, bolus, patch, external preparation for skin, bath preparation, heating device and the like. .
Preferably, the medicament or quasi-drug is a dentifrice, mouthwash, mouthspray, mouthwash, oral ointment, cream or the like containing the compound of formula (I) or formula (II) as an active ingredient. It can be an oral composition such as a coating agent. Also preferably, the oral composition is a composition for preventing and / or improving periodontal disease.
Preferably, the medicine or quasi-drug may be a patch, an external preparation for skin, a bath agent, or a heating tool containing the compound of formula (I) or formula (II) as an active ingredient.

  The medicament or quasi-drug may contain the compound of formula (I) or formula (II) 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, activity enhancers, anti-inflammatory agents, bactericides, flavoring agents, 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 responsiveness promoting effect of the compound of Formula (I) or Formula (II) is not lost.

  The pharmaceutical or quasi-drug is produced from the compound of formula (I) or formula (II) or, if necessary, by combining the carrier and / or other active ingredients and pharmacological ingredients according to a conventional method. Can do. The content of the compound of formula (I) or formula (II) in the pharmaceutical or quasi drug is preferably 0.0000001 to 100% by mass as the total of the compounds of formula (I) or formula (II). , 0.000001-100 mass% is more preferable, 0.000002-100 mass% is further preferable, 0.000005-100 mass% is still more preferable, 0.00001-100 mass% % Is still more preferable.

  The said food-drinks contain the compound of Formula (I) or Formula (II) as an active ingredient. The food or drink is based on the concept of promoting CGRP responsiveness, preventing and / or improving periodontal disease, and promoting blood flow and / or blood circulation. It can be set as health food, food and drink for the sick, and food and drink for specified health use. These foods and drinks can be distinguished from general foods and drinks by display.

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 preferable form, the food and drink of the present invention may be gum, candy, tablet, beverage, pet gum, and the like based on the concept of preventing and / or improving periodontal disease, promoting blood flow and / or blood circulation.

In order to prepare the above-mentioned various forms of foods and drinks, the compound of formula (I) or formula (II) alone or other food or 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 compound of formula (I) or formula (II) in the above food or drink is preferably 0.0000001 to 100% by mass in total of the compounds of formula (I) or formula (II). It is more preferable to set it as -75 mass%, It is further more preferable to set it as 0.000001-50 mass%, It is still more preferable to set it as 0.000001-30 mass%, It is set as 0.000001-10 mass%. Is still preferred.

  The cosmetic contains the compound of formula (I) or formula (II) as an active ingredient. The cosmetic may contain the compound of formula (I) or formula (II) alone or 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, thickeners, 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 compound of the formula (I) or the formula (II) is not lost, the cosmetic is other active ingredients and cosmetic ingredients, for example, a moisturizer, a whitening agent, an ultraviolet protective agent, It may contain cell activators, detergents, keratolytic agents, makeup ingredients (for example, makeup bases, foundations, funny, powder, teak, lipstick, eye makeup, eyebrow, mascara, etc.). Examples of the cosmetic preparation include creams, emulsions, lotions, suspensions, gels, powders, packs, sheets, patches, sticks, cakes, bathing agents, and any other form that can be used in cosmetics. .

  The cosmetic can be produced by a conventional method from the compound of formula (I) or formula (II) or, if necessary, in combination with the carrier and / or other active ingredients and cosmetic ingredients. The content of the compound of formula (I) or formula (II) in the cosmetic is preferably 0.0000001 to 100% by mass in total of the compounds of formula (I) or formula (II). It is more preferably 75% by mass, even more preferably 0.000001-50% by mass, still more preferably 0.000001-30% by mass, and 0.000001-10% by mass. Further preferred.

Alternatively, the compound of formula (I) or formula (II) is used for promoting CGRP responsiveness, for improving skin circulation or skin metabolism, for promoting blood flow and / or blood circulation, for promoting wound healing. Or high blood pressure, 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 delivery In order to prevent and / or ameliorate diseases such as these, they can be administered or ingested in an effective amount to a subject who needs or desires them.
In one embodiment, the administration or ingestion is performed non-therapeutically with the aim of improving skin circulation, improving skin metabolism, promoting blood flow and / or blood circulation for promoting health or cosmetic purposes.

  As an object of administration or ingestion, it is necessary or desirable to improve the reduction of skin metabolism due to aging or the like, preferably an animal that requires or desires improvement of skin circulation or skin metabolism. Include animals. Alternatively, the subjects of administration or ingestion include animals that need or desire blood flow and / or blood circulation or wound healing, as well as hypertension, heart failure, liver damage due to ischemic reperfusion, kidney Animals with or suspected to have illness, gastrointestinal ulcer, sepsis, osteoporosis, arrhythmia, subarachnoid hemorrhage, pulmonary hypertension, delayed hypersensitivity, periodontal disease, pregnancy toxemia, premature labor, etc. Or animals at high risk. Preferably, an animal suffering from periodontal disease, an animal suspected or high-risk thereof, or an animal in need of promotion of dental pulp formation is exemplified as a subject of administration or ingestion. The animal is preferably a human or non-human mammal, more preferably a human.

  The preferred dosage or intake may vary according to the subject's species, weight, sex, age, condition or other factors. The dose or route of administration or ingestion, the interval, and the amount or interval of ingestion can be appropriately determined by those skilled in the art. For example, when orally administered to humans, the dose is preferably 1 μg to 6000 mg / day per adult, and more preferably 10 μg to 5000 mg / day, as the sum of the compounds of formula (I) or formula (II). Preferably, 100 μg to 4000 mg / day is more preferable, and 500 μg to 3000 mg / day is still more preferable.

  Alternatively, the administration subject can be an animal-derived tissue, organ, cell, or a culture 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. Accordingly, the present invention also provides a method for promoting CGRP responsiveness in the tissue, organ, cell, or culture or fraction thereof. The method includes, for example, culturing a cell having a CGRP receptor and promoting CGRP responsiveness in the presence of a compound of formula (I) or formula (II). When the cell is a cell culture, the concentration of the compound of formula (I) or formula (II) added is 0.001 to 1000 μM in total of the compound of formula (I) or formula (II), preferably It is 0.002-500 micromol, More preferably, it is 0.01-200 micromol. The timing of adding the compound of formula (I) or formula (II) may be before CGRP stimulation and / or during CGRP stimulation.

  As exemplary embodiments of the present invention, the following compositions, manufacturing methods, uses or methods are further disclosed herein. However, the present invention is not limited to these embodiments.

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

Example 1
The effect of promoting the CGRP response of cells by the ginsenoside compound was examined.
1. Method (1) Cell culture Normal human coronary artery smooth muscle cells (HCASMC; Kurabo Corp., cell lot # 01272) are grown in growth medium (basic medium HuMedia-SB2 500 mL, FBS 25 mL, hEGF 0.5 mL, hFGF-B 0.5 mL. , 0.5 mL of insulin and 0.5 mL of antibacterial agent; Kurabo Industries Co., Ltd.). For passage, 0.025% trypsin / 0.01% EDTA was used.

(2) Cell preparation Normal human coronary artery smooth muscle cells (HCASMC) 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 (basal medium HuMedia-SB2 in 500 mL) FBS 5 mL, heparin 0.5 mL, antibacterial agent 0.5 mL added; The medium was changed every other day and cultured for 7 days, and used for measurement of CGRP responsiveness.

(3) Measurement of CGRP responsiveness The cells contain a phosphodiesterase inhibitor (PDI) IBMX (3-Isobutyl-1-methylxanthine, 500 μM; Sigma) and Ro-20-1724 (100 μM; Wako Pure Chemical Industries, Ltd.) The plate was washed twice with a basal medium (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 penetrate into the cells. The medium was removed, a basal medium containing 30 μM or 100 μM ginsenoside compound, 0.5 nM CGRP, and a phosphodiesterase inhibitor was added, and the mixture was 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 CGRP responsiveness promoting ability of the ginsenoside compound was expressed by [cAMP] i when stimulated with only 0.5 nM CGRP as a reference (control), and the percentage was expressed as Improve Index.
(4) Test compound The following ginsenoside compounds were used for the test. Ginsenoside Rb3, Rd, Re, Rf, Rg1, protopanaxatriol, purchased from Carbosynth, Ginsenoside Rb1, Rb2, Rc, Rg2, 20 (R) Rg3, 20 (S) Rg3, Rh1, Rh2, RF1, Protopanaxadiol was purchased from LKT Laboratories. Ginsenoside Rh3 was purchased from Cfm Oskar Tropitzsch, Ginsenoside Rk2 and Rs3 from WuXi App Tec, Ginsenoside F2 from BOC Sciences, and Ginsenoside Rg5 from WuXi App Tec.

2. Results The measurement results of CGRP responsiveness are shown in FIGS. The Improve Index 100% in FIG. 1 corresponds to [cAMP] i of 1113.5 fmol / well. The Improve Index 100% in FIG. 2 corresponds to [cAMP] i of 1234.5 fmol / well. Ginsenoside Rb1, Rb2 and Rb3, ginsenoside Rc, ginsenoside Rd, ginsenoside (20 (S)) Rg3 and (20 (R)) Rg3, ginsenoside Rh2, protopanaxatriol, protopanaxadiol, ginsenoside Rh3, ginsenoside Rk2, ginsenoside Rs3, ginsenoside F2, and ginsenoside Rg5 statistically significantly promoted the CGRP responsiveness of cells compared to the control in both cases of addition of 30 μM and 100 μM (Dunnett test, N = 3).

Example 2
Various addition conditions of the ginsenoside compound were changed, and the CGRP responsiveness of the cells was examined.
1. Method HCASMC (Kurabo, cell lot # 01272) was prepared under the same conditions as in Example 1. Thereafter, a phosphodiesterase inhibitor and ginsenoside and / or CGRP (0.5 nM) were added to the culture under the conditions described in Table 3, followed by culturing. Thereafter, the cells were responsive to CGRP in the same manner as in Example 1. Was measured. As ginsenoside, Rh2 (30 μM), 20 (S) Rg3 (100 μM), or 20 (R) Rg3 (100 μM) was used.

2. Result The measurement result of CGRP responsiveness is shown in FIG. Improve Index 100% corresponds to [cAMP] i of 1189.3 fmol / well. Improve Index increased by adding ginsenoside during CGRP stimulation (condition 2). On the other hand, since the Improve Index was statistically significantly lower in the absence of CGRP (condition 3), the action of the ginsenoside compound was not a substitute for CGRP (for example, a CGRP receptor agonist), but CGRP and It has been shown to promote a series of responses elicited by binding to its receptor.

Claims (4)

CGRP responsive promoter of cells as an active ingredient one or more one of the lower hear compounds and their optical isomers.
Under hear compounds and accelerators of blood flow and / or blood circulation as an active ingredient one or more any of those optical isomers.
Wherein the cell is a vascular smooth muscle cell, enhancer according to claim 1, wherein. 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 the CGRP responsive promoter according to claim 1 . Method.